Pub Date : 2026-01-10DOI: 10.1016/j.jep.2026.121185
Ruiyi Liu , Zhangjie Wu , Ying Yin , Chenghao Song , Chengyu Zhang , Shan Xing , Jingwen Tan , Zhenzhen Fu , Nga Lee Wong , Mingxue Deng , Mingzhi Han , Yuhan Peng , Changyu Wu , Huijun Xie , Gang Chen
Ethnopharmacological relevance
Insomnia and anxiety are highly comorbid, severely compromising quality of life. Efficacy of current pharmacological interventions for this dual condition remains limited. Zhi-Gan Formula (ZG), consisting of Zhi-Zi-Chi Decoction and Ganmai Dazao Decoction, two classic Traditional Chinese Medicine (TCM) formulae clinically widely used for insomnia or anxiety, holds promise as a therapeutic option for insomnia-anxiety comorbidity.
Aim of the study
This study aimed to assess ZG's sleep-promoting and anxiolytic efficacy, and investigate the novel mechanism through which pituitary adenylate cyclase-activating polypeptide (PACAP) in the medial prefrontal cortex (mPFC) modulates comorbid sleep and anxiety conditions.
Materials and methods
Mice received 4-chloro-DL-phenylalanine (PCPA) injections and were subsequently administered ZG or diazepam. Behaviors were assessed using the pentobarbital-induced sleep test, open-field test (OFT), and elevated plus-maze test (EPM). Key pathways were identified via network pharmacology analysis and validated using long-term potentiation (LTP) recordings and protein quantification. Viral-mediated PACAP knockdown vectors were transfected into the mPFC.
Results
PCPA administration induced insomnia and anxiety-like behaviors. ZG administered for 3 days significantly shortened sleep latency, prolonged sleep duration, and alleviated anxiety-like behaviors, whereas diazepam only partially improved anxiety-like behaviors. Network pharmacology analysis suggested ZG's engagement in neuropeptide-receptor interactions and synaptic transmission pathways. Assessments of synaptic plasticity showed that ZG improved mPFC LTP and the expression of synaptic proteins (PSD95, synapsin-1, BDNF) impaired in the model mice. Moreover, the expression of the neuropeptide PACAP and downstream eEF2 signaling for synaptic protein synthesis were all improved by ZG. Crucially, perfusion of a PACAP agonist in the mPFC brain slices from sleep-deprived mice rescued LTP deficits. Finally, mPFC PACAP knockdown abolished the therapeutic effects and the enhanced expressions of the synaptic proteins by ZG.
Conclusions
ZG alleviated insomnia-anxiety comorbidity by restoring synaptic plasticity in the mPFC via the PACAP-eEF2-BDNF pathway, which may also shed light on the development of a novel therapeutic approach for the treatment of sleep-anxiety comorbidity.
{"title":"Zhi-Gan Formula improved insomnia and anxiety comorbidity in a mouse model via PACAP signaling in the medial prefrontal cortex","authors":"Ruiyi Liu , Zhangjie Wu , Ying Yin , Chenghao Song , Chengyu Zhang , Shan Xing , Jingwen Tan , Zhenzhen Fu , Nga Lee Wong , Mingxue Deng , Mingzhi Han , Yuhan Peng , Changyu Wu , Huijun Xie , Gang Chen","doi":"10.1016/j.jep.2026.121185","DOIUrl":"10.1016/j.jep.2026.121185","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div>Insomnia and anxiety are highly comorbid, severely compromising quality of life. Efficacy of current pharmacological interventions for this dual condition remains limited. Zhi-Gan Formula (ZG), consisting of Zhi-Zi-Chi Decoction and Ganmai Dazao Decoction, two classic Traditional Chinese Medicine (TCM) formulae clinically widely used for insomnia or anxiety, holds promise as a therapeutic option for insomnia-anxiety comorbidity.</div></div><div><h3>Aim of the study</h3><div>This study aimed to assess ZG's sleep-promoting and anxiolytic efficacy, and investigate the novel mechanism through which pituitary adenylate cyclase-activating polypeptide (PACAP) in the medial prefrontal cortex (mPFC) modulates comorbid sleep and anxiety conditions.</div></div><div><h3>Materials and methods</h3><div>Mice received 4-chloro-DL-phenylalanine (PCPA) injections and were subsequently administered ZG or diazepam. Behaviors were assessed using the pentobarbital-induced sleep test, open-field test (OFT), and elevated plus-maze test (EPM). Key pathways were identified via network pharmacology analysis and validated using long-term potentiation (LTP) recordings and protein quantification. Viral-mediated PACAP knockdown vectors were transfected into the mPFC.</div></div><div><h3>Results</h3><div>PCPA administration induced insomnia and anxiety-like behaviors. ZG administered for 3 days significantly shortened sleep latency, prolonged sleep duration, and alleviated anxiety-like behaviors, whereas diazepam only partially improved anxiety-like behaviors. Network pharmacology analysis suggested ZG's engagement in neuropeptide-receptor interactions and synaptic transmission pathways. Assessments of synaptic plasticity showed that ZG improved mPFC LTP and the expression of synaptic proteins (PSD95, synapsin-1, BDNF) impaired in the model mice. Moreover, the expression of the neuropeptide PACAP and downstream eEF2 signaling for synaptic protein synthesis were all improved by ZG. Crucially, perfusion of a PACAP agonist in the mPFC brain slices from sleep-deprived mice rescued LTP deficits. Finally, mPFC PACAP knockdown abolished the therapeutic effects and the enhanced expressions of the synaptic proteins by ZG.</div></div><div><h3>Conclusions</h3><div>ZG alleviated insomnia-anxiety comorbidity by restoring synaptic plasticity in the mPFC via the PACAP-eEF2-BDNF pathway, which may also shed light on the development of a novel therapeutic approach for the treatment of sleep-anxiety comorbidity.</div></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121185"},"PeriodicalIF":5.4,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145958771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-09DOI: 10.1016/j.jep.2026.121176
Wenwen Zhu , Ranran Chen , Lianhao Wu , Yiyang Shui , Longhai Liu , Junyi Wang , Yi Sun , Yafei Yu , Yu Yuan , Zhenqiang You , Yueguo Wu
Ethnopharmacological relevance
Dendrobium officinale Kimura et Migo (D. officinale), a rare and precious orchid (Orchidaceae) in traditional Chinese medicine, exhibits a sweet flavor and a cold nature, and acts on lung, stomach, and kidney meridians with marked efficacy in nourishing lung yin. When applied medicinally and in functional foods, it demonstrates antioxidant, immunomodulatory, anti-inflammatory, and anti-inflammatory, and antitumor bioactivities.
Aim of the study
This study aimed to investigate the role and mechanisms of action of an ethyl acetate extract of D. officinale (DOEA) in the treatment of acute lung injury (ALI).
Materials and methods
The pharmacological effects of DOEA extract were evaluated using lipopolysaccharide (LPS)-induced ALI models in vivo and RAW264.7 cell models in vitro. Transcriptomic profiling was performed via RNA sequencing to elucidate the mechanism by which DOEA protects against LPS-induced injury. ALI Furthermore, the protective effects of DOEA against LPS-induced ALI and cellular inflammatory responses were comprehensively evaluated through histology, lung index assessment, immunofluorescence, immunohistochemistry, Western blotting, quantitative real-time PCR, and flow cytometry. Exploring the anti-inflammatory effects of DOEA on the IL-17A signaling pathway was achieved by knocking down IL-17RA in RAW264.7 cells via siRNA interference technology, as well as by conducting exogenous IL-17A stimulation experiments.
Results
DOEA mitigated LPS-induced ALI by attenuating pulmonary cytokine expression, suppressing NF-κB pathway activation, and reducing oxidative tissue damage via the inhibition of cellular oxidative stress. Moreover, the efficacy of DOEA against ALI was significantly associated with the targeting of IL-17RA. Notably, IL-17RA knockdown significantly alleviated the LPS-induced inflammatory response in RAW264.7 cells. Furthermore, DOEA was able to suppress the inflammatory response induced by exogenous IL-17A stimulation in these cells.
Conclusion
DOEA alleviates LPS-induced acute lung injury and cellular inflammatory responses by inhibiting the IL-17 signaling pathway.
民族药理学相关性：木村石斛（Dendrobium officinale Kimura et Migo）是一种稀有珍贵的中药兰花（兰科），味甘性寒，作用于肺、胃、肾经，具有显著的养肺阴功效。当药用和功能性食品应用时，它显示出抗氧化、免疫调节、抗炎、抗炎和抗肿瘤的生物活性。研究目的：探讨officinale乙酸乙酯提取物（DOEA）对急性肺损伤（ALI）的治疗作用及其机制。材料与方法：采用脂多糖（LPS）诱导的体内ALI模型和体外RAW264.7细胞模型，评价DOEA提取物的药理作用。转录组学分析通过RNA测序来阐明DOEA对lps诱导的损伤的保护机制。进一步，通过组织学、肺指数评估、免疫荧光、免疫组织化学、Western blotting、实时荧光定量PCR、流式细胞术等方法，综合评价DOEA对lps诱导的ALI及细胞炎症反应的保护作用。通过siRNA干扰技术敲除RAW264.7细胞中的IL-17RA，并进行外源性IL-17A刺激实验，探索DOEA对IL-17A信号通路的抗炎作用。结果：DOEA通过降低肺细胞因子表达，抑制NF-κB通路激活，通过抑制细胞氧化应激减轻氧化组织损伤，减轻lps诱导的ALI。此外，DOEA对ALI的疗效与IL-17RA的靶向性显著相关。值得注意的是，IL-17RA敲低显著减轻了lps诱导的RAW264.7细胞炎症反应。此外，DOEA能够抑制外源性IL-17A刺激引起的这些细胞的炎症反应。结论：DOEA通过抑制IL-17信号通路减轻lps诱导的急性肺损伤和细胞炎症反应。
{"title":"Ethyl acetate fraction from Dendrobium officinale inhibits IL-17A signaling pathway to mitigate acute lung injury","authors":"Wenwen Zhu , Ranran Chen , Lianhao Wu , Yiyang Shui , Longhai Liu , Junyi Wang , Yi Sun , Yafei Yu , Yu Yuan , Zhenqiang You , Yueguo Wu","doi":"10.1016/j.jep.2026.121176","DOIUrl":"10.1016/j.jep.2026.121176","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div><em>Dendrobium officinale</em> Kimura et Migo (<em>D. officinale</em>), a rare and precious orchid (Orchidaceae) in traditional Chinese medicine, exhibits a sweet flavor and a cold nature, and acts on lung, stomach, and kidney meridians with marked efficacy in nourishing lung yin. When applied medicinally and in functional foods, it demonstrates antioxidant, immunomodulatory, anti-inflammatory, and anti-inflammatory, and antitumor bioactivities.</div></div><div><h3>Aim of the study</h3><div>This study aimed to investigate the role and mechanisms of action of an ethyl acetate extract of <em>D. officinale</em> (DOEA) in the treatment of acute lung injury (ALI).</div></div><div><h3>Materials and methods</h3><div>The pharmacological effects of DOEA extract were evaluated using lipopolysaccharide (LPS)-induced ALI models in vivo and RAW264.7 cell models in vitro. Transcriptomic profiling was performed via RNA sequencing to elucidate the mechanism by which DOEA protects against LPS-induced injury. ALI Furthermore, the protective effects of DOEA against LPS-induced ALI and cellular inflammatory responses were comprehensively evaluated through histology, lung index assessment, immunofluorescence, immunohistochemistry, Western blotting, quantitative real-time PCR, and flow cytometry. Exploring the anti-inflammatory effects of DOEA on the IL-17A signaling pathway was achieved by knocking down IL-17RA in RAW264.7 cells via siRNA interference technology, as well as by conducting exogenous IL-17A stimulation experiments.</div></div><div><h3>Results</h3><div>DOEA mitigated LPS-induced ALI by attenuating pulmonary cytokine expression, suppressing NF-κB pathway activation, and reducing oxidative tissue damage via the inhibition of cellular oxidative stress. Moreover, the efficacy of DOEA against ALI was significantly associated with the targeting of IL-17RA. Notably, IL-17RA knockdown significantly alleviated the LPS-induced inflammatory response in RAW264.7 cells. Furthermore, DOEA was able to suppress the inflammatory response induced by exogenous IL-17A stimulation in these cells.</div></div><div><h3>Conclusion</h3><div>DOEA alleviates LPS-induced acute lung injury and cellular inflammatory responses by inhibiting the IL-17 signaling pathway.</div></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121176"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tetrastigma hemsleyanum Diels et Gilg, traditionally used in southern China to treat fever, cough, and pneumonia, is a well-known ethnomedicinal plant. Polysaccharides extracted from its aboveground parts (leaves and canes) possess potent immunomodulatory and anti-inflammatory activities. However, the mechanisms underlying their protective effects against viral-mimic acute lung injury (ALI) remain largely unexplored.
Materials and methods
Mice were pretreated with STHP-5 (1–4 mg/kg, intratracheal instillation) once daily for three days before Poly(I:C) challenge to induce ALI. Lung injury, inflammation, and barrier integrity were evaluated by histopathology (H&E), TUNEL assay, wet/dry ratio, bronchoalveolar lavage fluid (BALF) analysis, myeloperoxidase (MPO) staining, and flow cytometry of immune cells. Cytokine levels, oxidative stress, and tight-junction proteins were assessed by cytometric bead array (CBA), ROS detection, Western blotting, and quantitative real-time PCR (RT-qPCR). In vitro, bone marrow–derived macrophages (BMDMs) and BEAS-2B epithelial cells were stimulated with Poly(I:C) with or without STHP-5 pretreatment, and examined by RT-qPCR, Western blotting, immunofluorescence, and cytokine assays to evaluate the STING/TBK1/NF-κB and IRF1/STAT1 signaling axes.
Results
Prophylactic administration of STHP-5 significantly alleviated pulmonary inflammation, reduced cytokine secretion, and preserved alveolar epithelial integrity in Poly(I:C)-challenged mice. Further analysis revealed that STHP-5 inhibited activation of the STING/TBK1/NF-κB and IRF1/STAT1 signaling pathways in BMDMs, thereby suppressing M1 macrophage polarization and pro-inflammatory cytokine production. Additionally, STHP-5 protected BEAS-2B cells from Poly(I:C)-induced injury by maintaining tight-junction proteins and reducing oxidative stress.
Conclusion
STHP-5 effectively mitigates Poly(I:C)-induced acute lung injury through coordinated suppression of innate immune hyperactivation and reinforcement of epithelial barrier defense. These findings provide new mechanistic insight into the multi-target pharmacological actions of STHP-5 and support its potential development as a safe, locally deliverable polysaccharide-based therapeutic for viral and inflammatory lung diseases.
{"title":"Polysaccharide from Tetrastigma hemsleyanum Diels et Gilg attenuates Poly(I:C)-induced acute lung injury by preserving epithelial barrier integrity and modulating STING/TBK1–NF-κB and IRF1–STAT1 signaling","authors":"Mengjia Zhao , Cong Jin , Jingwen Xu , Haotian Wang , Shuting Huang , Kuixu Gao , Zhixi Lin , Shengjun Chen , Yuchi Chen , Fangmei Zhou , Bingqi Zhu , Zhishan Ding , Mingyuan Zhou","doi":"10.1016/j.jep.2026.121192","DOIUrl":"10.1016/j.jep.2026.121192","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div><em>Tetrastigma hemsleyanum</em> Diels et Gilg, traditionally used in southern China to treat fever, cough, and pneumonia, is a well-known ethnomedicinal plant. Polysaccharides extracted from its aboveground parts (leaves and canes) possess potent immunomodulatory and anti-inflammatory activities. However, the mechanisms underlying their protective effects against viral-mimic acute lung injury (ALI) remain largely unexplored.</div></div><div><h3>Materials and methods</h3><div>Mice were pretreated with STHP-5 (1–4 mg/kg, intratracheal instillation) once daily for three days before Poly(I:C) challenge to induce ALI. Lung injury, inflammation, and barrier integrity were evaluated by histopathology (H&E), TUNEL assay, wet/dry ratio, bronchoalveolar lavage fluid (BALF) analysis, myeloperoxidase (MPO) staining, and flow cytometry of immune cells. Cytokine levels, oxidative stress, and tight-junction proteins were assessed by cytometric bead array (CBA), ROS detection, Western blotting, and quantitative real-time PCR (RT-qPCR). <em>In vitro</em>, bone marrow–derived macrophages (BMDMs) and BEAS-2B epithelial cells were stimulated with Poly(I:C) with or without STHP-5 pretreatment, and examined by RT-qPCR, Western blotting, immunofluorescence, and cytokine assays to evaluate the STING/TBK1/NF-κB and IRF1/STAT1 signaling axes.</div></div><div><h3>Results</h3><div>Prophylactic administration of STHP-5 significantly alleviated pulmonary inflammation, reduced cytokine secretion, and preserved alveolar epithelial integrity in Poly(I:C)-challenged mice. Further analysis revealed that STHP-5 inhibited activation of the STING/TBK1/NF-κB and IRF1/STAT1 signaling pathways in BMDMs, thereby suppressing M1 macrophage polarization and pro-inflammatory cytokine production. Additionally, STHP-5 protected BEAS-2B cells from Poly(I:C)-induced injury by maintaining tight-junction proteins and reducing oxidative stress.</div></div><div><h3>Conclusion</h3><div>STHP-5 effectively mitigates Poly(I:C)-induced acute lung injury through coordinated suppression of innate immune hyperactivation and reinforcement of epithelial barrier defense. These findings provide new mechanistic insight into the multi-target pharmacological actions of STHP-5 and support its potential development as a safe, locally deliverable polysaccharide-based therapeutic for viral and inflammatory lung diseases.</div></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121192"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<div><h3>Ethnopharmacological relevance</h3><div>According to a report published in 2025, Cameroon has the highest breast cancer mortality rate in Africa, with 91,300 deaths recorded in 2022 out of 198,300 new cases. Referring to the use of <em>Acacia seyal</em> Delile in Cameroonian traditional system to treat inflammatory conditions and cancer, we demonstrated that its hydro-ethanolic stem barks extract has anti-breast cancer potential <em>in vitro</em> and <em>in vivo</em>.</div><div><em>Aim of the study</em>: To evaluate the modes of action (anti-oxidant, anti-inflammatory and anti-estrogenic activities) of fractions (polyphenols, polysaccharides and residue) from <em>Acacia seyal</em> Delile.</div></div><div><h3>Materials and methods</h3><div>Once the polyphenol-rich (ASpo), polysaccharides (ASsu) and residue (ASres) fractions were obtained, their total phenolic, and UPLC-HR-LC-MS fingerprints were determined and compared to those of the crude extract. Their <em>in vitro</em> anti-oxidant (DPPH, ABTS and total anti-oxidant capacity), anti-inflammatory (egg albumin denaturation and human erythrocyte membrane stabilization) and estrogenic/anti-estrogenic (MCF-7 cell proliferation/E-screen) activities were determined. Having demonstrated the best activities, the ASpo fraction was then evaluated for its <em>in vivo</em> anti-oxidant effect using the D-Galactose-induced oxidative stress rat model, anti-inflammatory activity via the carrageenan induced-inflammation rat model and estrogenic/anti-estrogenic activity through the 3-day uterotrophic assay in rats.</div></div><div><h3>Results</h3><div>The <em>Acacia seyal</em> Delile polyphenol-rich fraction (ASpo) showed a higher total phenol content (6292.53 ± 282.03 μg Eq ascorbic acid/mg dry extract) with its UHPLC chromatograms presenting many peaks putatively detected in the range of flavonoids. It was the most potent fraction with potent anti-oxidative activities (EC<sub>50</sub> of 0.05 mg/mL, 100 % free radical scavenging after 15 min in DPPH). It contents 2.9 mg Eq trolox/g dry matter of antiradical ingredients and 2803.92 ± 9.61 μg Eq ascorbic acid/g dry matter in ATBS and TAC, respectively. ASpo at 4 mg/mL had a highest percentage of membrane stabilization (97.9 %) and protected the egg albumin against denaturation induced by the heat with IC<sub>50</sub> of 70.2 μg/mL. ASpo exhibited <em>in vitro</em> estrogenic and anti-estrogenic effect in MCF-7 cells in dose-dependent fashion. <em>In vivo</em>, ASpo exhibited a significant and dose-dependent reduction of the MDA levels, increase the GSH levels and SOD activity with a maximum effect (<em>p</em> < 0.001) observed at 150 mg/kg BW. However, at a dose of 75 mg/kg, ASpo had a similar effect to that induced by the crude extract (ASEH) at a dose of 150 mg/kg BW. The ASpo exhibited a dose-dependent protection against hepatotoxicity and nephrotoxicity induced by the D-Galactose in rats, as well as a dose-dependent and time-dependent inhibition
{"title":"Anti-oxidant, anti-estrogenic and anti-inflammatory activities of the UHPLC fingerprinted Acacia seyal Delile (Mimosaceae) extracts: possible anticancer modes of action","authors":"Alain Brice Tueche , Kevine Silihe Kamga , Charlotte Mungoh Tata , Marius Trésor Kemegne Sipping , Calvin Zangueu Bogning , Edwin Mpoh Mmutlane , Alain Bertrand Dongmo , Dieudonné Njamen , Stéphane Zingue , Derek Tantoh Ndinteh","doi":"10.1016/j.jep.2026.121193","DOIUrl":"10.1016/j.jep.2026.121193","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div>According to a report published in 2025, Cameroon has the highest breast cancer mortality rate in Africa, with 91,300 deaths recorded in 2022 out of 198,300 new cases. Referring to the use of <em>Acacia seyal</em> Delile in Cameroonian traditional system to treat inflammatory conditions and cancer, we demonstrated that its hydro-ethanolic stem barks extract has anti-breast cancer potential <em>in vitro</em> and <em>in vivo</em>.</div><div><em>Aim of the study</em>: To evaluate the modes of action (anti-oxidant, anti-inflammatory and anti-estrogenic activities) of fractions (polyphenols, polysaccharides and residue) from <em>Acacia seyal</em> Delile.</div></div><div><h3>Materials and methods</h3><div>Once the polyphenol-rich (ASpo), polysaccharides (ASsu) and residue (ASres) fractions were obtained, their total phenolic, and UPLC-HR-LC-MS fingerprints were determined and compared to those of the crude extract. Their <em>in vitro</em> anti-oxidant (DPPH, ABTS and total anti-oxidant capacity), anti-inflammatory (egg albumin denaturation and human erythrocyte membrane stabilization) and estrogenic/anti-estrogenic (MCF-7 cell proliferation/E-screen) activities were determined. Having demonstrated the best activities, the ASpo fraction was then evaluated for its <em>in vivo</em> anti-oxidant effect using the D-Galactose-induced oxidative stress rat model, anti-inflammatory activity via the carrageenan induced-inflammation rat model and estrogenic/anti-estrogenic activity through the 3-day uterotrophic assay in rats.</div></div><div><h3>Results</h3><div>The <em>Acacia seyal</em> Delile polyphenol-rich fraction (ASpo) showed a higher total phenol content (6292.53 ± 282.03 μg Eq ascorbic acid/mg dry extract) with its UHPLC chromatograms presenting many peaks putatively detected in the range of flavonoids. It was the most potent fraction with potent anti-oxidative activities (EC<sub>50</sub> of 0.05 mg/mL, 100 % free radical scavenging after 15 min in DPPH). It contents 2.9 mg Eq trolox/g dry matter of antiradical ingredients and 2803.92 ± 9.61 μg Eq ascorbic acid/g dry matter in ATBS and TAC, respectively. ASpo at 4 mg/mL had a highest percentage of membrane stabilization (97.9 %) and protected the egg albumin against denaturation induced by the heat with IC<sub>50</sub> of 70.2 μg/mL. ASpo exhibited <em>in vitro</em> estrogenic and anti-estrogenic effect in MCF-7 cells in dose-dependent fashion. <em>In vivo</em>, ASpo exhibited a significant and dose-dependent reduction of the MDA levels, increase the GSH levels and SOD activity with a maximum effect (<em>p</em> < 0.001) observed at 150 mg/kg BW. However, at a dose of 75 mg/kg, ASpo had a similar effect to that induced by the crude extract (ASEH) at a dose of 150 mg/kg BW. The ASpo exhibited a dose-dependent protection against hepatotoxicity and nephrotoxicity induced by the D-Galactose in rats, as well as a dose-dependent and time-dependent inhibition ","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121193"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952246","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-09DOI: 10.1016/j.jep.2026.121183
Xiaoying Zhang , Sha Xiang , Siyun Tang , Xiaoxia Luo , Lingyang Fan , Xianlei Fang , Hua Chen , Zaiqi Zhang , Kai He
<div><h3>Ethnopharmacological relevance</h3><div>The bark of <em>Myrica rubra</em> (Lour.) Siebold & Zucc (MR). is a natural remedy commonly used in China and other Asian nations because of its antioxidant, antiinflammation and antibacterial activities. Myricanol is the main lipid-lowering compounds in the bark of MR. The effects of myricanol on alleviating hyperlipidemia have rarely been reported. No study has investigated the role of the ACSL-SCD axis in the management of hyperlipidemia in vivo.</div></div><div><h3>Aim of the study</h3><div>This study employed a high-fat diet (HFD)-induced hyperlipidemic C57BL/6J mouse model to explore the lipid-lowering effects and underlying mechanisms of myricanol through the ACSL1-SCD1 axis.</div></div><div><h3>Materials and methods</h3><div>An olive oil and lard mixture was used to establish a hyperlipidemic C57BL/6J mouse model. Rosiglitazone (RSG) a peroxisome proliferator-activated receptor γ (PPARγ) agonist, and ACSL inhibitor was used as a positive control. After 8 weeks of high-fat modeling, the mice were randomly divided into the M group, RSG group (0.4 ml, 0.78 mg/kg RSG solution daily), low-dose myricanol group (MYL, 100 mg/kg), and high-dose myricanol group (MYH, 150 mg/kg). After 25 days of treatment, the lipid-lowering effects of myricanol were evaluated by measuring serum lipid levels and histopathological observation. Western blotting, metabolomics, 16S rRNA sequencing, RNA sequencing, ELISA, immunofluorescence staining, double-fluorescence labeling and cellular thermal shift assay (CETSA) were used to explore the underlying mechanisms.</div></div><div><h3>Results</h3><div>Only 7 days of myricanol treatment significantly reduced body weight in obese mice. Myricanol normalized serum levels of TC, TG, HDL-C, and LDL-C; reduced lipid droplet accumulation in hepatocytes; and decreased the epididymal fat volume in mice. Mechanistic studies revealed that myricanol upregulated the expression of ACSL1, PPARγ, CYP7A1, SCD1, ACLY, and SREBF1 in the mouse liver. Additionally, myricanol increased the expression of PPARγ, CPT1A, ACC1, ACSL1, APOE4, and SREBF1 in the mouse epididymal fat. Multimodal omics analyses indicated that the lipid-lowering activity of myricanol was partially mediated by modulating the gut microbiota, such as that of <em>Monoglobus</em> and <em>Lachnospiraceae bacterium 28-4</em>; regulating the interferon (IFN) pathway and IFN-stimulated genes; and influencing the expression of miRNAs such as miR-203b-3p, miR-205-5p, and miR-184-3p. Cellular thermal shift assay, molecular docking, and ELISA confirmed that myricanol directly bound to ACSL1 and decreased the concentrations of ACSL1 and SCD1 in mouse serum. Intriguingly, myricanol promoted mitochondrial biogenesis in a time- and dose-dependent manner and increased ketone body and acetyl-CoA levels in obese mice.</div></div><div><h3>Conclusion</h3><div>This study reveals a novel cellular mechanism through which myricanol reduces li
{"title":"Myricanol modulates PPARγ/ACSL1/SCD1 metabolic signaling pathway to promote mitochondria biogenesis and fatty acid β-oxidation in high-fat diet-induced obese mice","authors":"Xiaoying Zhang , Sha Xiang , Siyun Tang , Xiaoxia Luo , Lingyang Fan , Xianlei Fang , Hua Chen , Zaiqi Zhang , Kai He","doi":"10.1016/j.jep.2026.121183","DOIUrl":"10.1016/j.jep.2026.121183","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div>The bark of <em>Myrica rubra</em> (Lour.) Siebold & Zucc (MR). is a natural remedy commonly used in China and other Asian nations because of its antioxidant, antiinflammation and antibacterial activities. Myricanol is the main lipid-lowering compounds in the bark of MR. The effects of myricanol on alleviating hyperlipidemia have rarely been reported. No study has investigated the role of the ACSL-SCD axis in the management of hyperlipidemia in vivo.</div></div><div><h3>Aim of the study</h3><div>This study employed a high-fat diet (HFD)-induced hyperlipidemic C57BL/6J mouse model to explore the lipid-lowering effects and underlying mechanisms of myricanol through the ACSL1-SCD1 axis.</div></div><div><h3>Materials and methods</h3><div>An olive oil and lard mixture was used to establish a hyperlipidemic C57BL/6J mouse model. Rosiglitazone (RSG) a peroxisome proliferator-activated receptor γ (PPARγ) agonist, and ACSL inhibitor was used as a positive control. After 8 weeks of high-fat modeling, the mice were randomly divided into the M group, RSG group (0.4 ml, 0.78 mg/kg RSG solution daily), low-dose myricanol group (MYL, 100 mg/kg), and high-dose myricanol group (MYH, 150 mg/kg). After 25 days of treatment, the lipid-lowering effects of myricanol were evaluated by measuring serum lipid levels and histopathological observation. Western blotting, metabolomics, 16S rRNA sequencing, RNA sequencing, ELISA, immunofluorescence staining, double-fluorescence labeling and cellular thermal shift assay (CETSA) were used to explore the underlying mechanisms.</div></div><div><h3>Results</h3><div>Only 7 days of myricanol treatment significantly reduced body weight in obese mice. Myricanol normalized serum levels of TC, TG, HDL-C, and LDL-C; reduced lipid droplet accumulation in hepatocytes; and decreased the epididymal fat volume in mice. Mechanistic studies revealed that myricanol upregulated the expression of ACSL1, PPARγ, CYP7A1, SCD1, ACLY, and SREBF1 in the mouse liver. Additionally, myricanol increased the expression of PPARγ, CPT1A, ACC1, ACSL1, APOE4, and SREBF1 in the mouse epididymal fat. Multimodal omics analyses indicated that the lipid-lowering activity of myricanol was partially mediated by modulating the gut microbiota, such as that of <em>Monoglobus</em> and <em>Lachnospiraceae bacterium 28-4</em>; regulating the interferon (IFN) pathway and IFN-stimulated genes; and influencing the expression of miRNAs such as miR-203b-3p, miR-205-5p, and miR-184-3p. Cellular thermal shift assay, molecular docking, and ELISA confirmed that myricanol directly bound to ACSL1 and decreased the concentrations of ACSL1 and SCD1 in mouse serum. Intriguingly, myricanol promoted mitochondrial biogenesis in a time- and dose-dependent manner and increased ketone body and acetyl-CoA levels in obese mice.</div></div><div><h3>Conclusion</h3><div>This study reveals a novel cellular mechanism through which myricanol reduces li","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121183"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-09DOI: 10.1016/j.jep.2026.121179
Le Viet Ha Tran , Huu Canh Vo , Tran Dang Linh Nguyen , To Hoang Long , Minh-Tri Le , Khac-Minh Thai , Truc Ly Nguyen , Van-Kieu Nguyen , Huynh Nguyen Khanh Tran
<div><h3>Ethnobotanical relevance</h3><div><em>Morus alba</em> L. has been reported to exhibit various pharmacological effects, including antipyretic, hepatoprotective, nephroprotective, antihypertensive, and anti-inflammatory activities, as well as benefits for sore throat relief and eyesight improvement in traditional medicine.</div></div><div><h3>Aim of the study</h3><div>This study investigates the anti-inflammatory potential of two prenylated flavonoids isolated from the twigs of <em>Morus alba</em> L.</div></div><div><h3>Methods</h3><div>The methanol extract and ethyl acetate (EA)-soluble fraction exhibited inhibitory effects on nitric oxide (NO) production. These fractions were further purified using repeated open-column chromatographic techniques, including liquid–liquid partitioning, normal-phase (NP) and reversed-phase (RP) chromatography, Sephadex gel filtration, and preparative HPLC. The chemical structures of compounds <strong>1</strong> and <strong>2</strong> were elucidated based on comprehensive spectroscopic analyses, including 1D and 2D NMR, experimental circular dichroism (CD), and specific optical rotation, along with comparison to reported data in the literature. The inhibitory effects on NO production and cell viability were evaluated in RAW 264.7 macrophage cells using the MTT assay. In addition, molecular docking and pharmacokinetic property predictions were performed for the two promising compounds.</div></div><div><h3>Results</h3><div>The methanol extract and ethyl acetate (EA)-soluble fraction exhibited inhibitory effects on nitric oxide (NO) production, showing 65 % inhibition at 100 μg/mL. These fractions underwent bioassay-guided fractionation, leading to the isolation of morusoin A (<strong>1</strong>) and (−)-mulberranol (<strong>2</strong>), two bioactive prenylated flavonoids. Compounds <strong>1</strong> and <strong>2</strong> significantly inhibited NO production in a dose-dependent manner without affecting cell viability. Furthermore, molecular docking studies were performed to evaluate the interactions of these two flavonoids with key pro-inflammatory cytokines and transcription factors (iNOS, COX-2, TNF-α, IL-1β, IL-6, Nrf2, and NF-κB) to elucidate their potential anti-inflammatory mechanisms. The binding energies were calculated as −9.0/−8.0, −9.1/−9.6, −6.0/−6.2, −6.0/−5.6, −6.5/−6.8, −7.1/−9.4, and −8.8/−9.5 kcal/mol, respectively. In addition, <em>in silico</em> ADMET and toxicity analyses were performed, providing an overview of the pharmacokinetic and safety profiles of the two compounds.</div></div><div><h3>Conclusion</h3><div>Compounds <strong>1</strong> and <strong>2</strong> have been well studied for isolation, structural elucidation, and their anti-inflammatory effects using both <em>in vitro</em> and <em>in silico</em> approaches and indicated that <strong>1</strong> and <strong>2</strong> possessed potential anti-inflammatory activity mediated via the Nrf2/NF-κB signaling pathway, warranting su
{"title":"Isolation, structural re-elucidation of two active prenylated flavonoids from Morus alba L. twigs responsible for anti-inflammatory effects: an in vitro and in silico approach","authors":"Le Viet Ha Tran , Huu Canh Vo , Tran Dang Linh Nguyen , To Hoang Long , Minh-Tri Le , Khac-Minh Thai , Truc Ly Nguyen , Van-Kieu Nguyen , Huynh Nguyen Khanh Tran","doi":"10.1016/j.jep.2026.121179","DOIUrl":"10.1016/j.jep.2026.121179","url":null,"abstract":"<div><h3>Ethnobotanical relevance</h3><div><em>Morus alba</em> L. has been reported to exhibit various pharmacological effects, including antipyretic, hepatoprotective, nephroprotective, antihypertensive, and anti-inflammatory activities, as well as benefits for sore throat relief and eyesight improvement in traditional medicine.</div></div><div><h3>Aim of the study</h3><div>This study investigates the anti-inflammatory potential of two prenylated flavonoids isolated from the twigs of <em>Morus alba</em> L.</div></div><div><h3>Methods</h3><div>The methanol extract and ethyl acetate (EA)-soluble fraction exhibited inhibitory effects on nitric oxide (NO) production. These fractions were further purified using repeated open-column chromatographic techniques, including liquid–liquid partitioning, normal-phase (NP) and reversed-phase (RP) chromatography, Sephadex gel filtration, and preparative HPLC. The chemical structures of compounds <strong>1</strong> and <strong>2</strong> were elucidated based on comprehensive spectroscopic analyses, including 1D and 2D NMR, experimental circular dichroism (CD), and specific optical rotation, along with comparison to reported data in the literature. The inhibitory effects on NO production and cell viability were evaluated in RAW 264.7 macrophage cells using the MTT assay. In addition, molecular docking and pharmacokinetic property predictions were performed for the two promising compounds.</div></div><div><h3>Results</h3><div>The methanol extract and ethyl acetate (EA)-soluble fraction exhibited inhibitory effects on nitric oxide (NO) production, showing 65 % inhibition at 100 μg/mL. These fractions underwent bioassay-guided fractionation, leading to the isolation of morusoin A (<strong>1</strong>) and (−)-mulberranol (<strong>2</strong>), two bioactive prenylated flavonoids. Compounds <strong>1</strong> and <strong>2</strong> significantly inhibited NO production in a dose-dependent manner without affecting cell viability. Furthermore, molecular docking studies were performed to evaluate the interactions of these two flavonoids with key pro-inflammatory cytokines and transcription factors (iNOS, COX-2, TNF-α, IL-1β, IL-6, Nrf2, and NF-κB) to elucidate their potential anti-inflammatory mechanisms. The binding energies were calculated as −9.0/−8.0, −9.1/−9.6, −6.0/−6.2, −6.0/−5.6, −6.5/−6.8, −7.1/−9.4, and −8.8/−9.5 kcal/mol, respectively. In addition, <em>in silico</em> ADMET and toxicity analyses were performed, providing an overview of the pharmacokinetic and safety profiles of the two compounds.</div></div><div><h3>Conclusion</h3><div>Compounds <strong>1</strong> and <strong>2</strong> have been well studied for isolation, structural elucidation, and their anti-inflammatory effects using both <em>in vitro</em> and <em>in silico</em> approaches and indicated that <strong>1</strong> and <strong>2</strong> possessed potential anti-inflammatory activity mediated via the Nrf2/NF-κB signaling pathway, warranting su","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121179"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-09DOI: 10.1016/j.jep.2026.121190
Lihao Chen , Jun Peng , Genyan Qin , Dongdong Li , Xiaolei Yao , Qinghua Peng
Ethnopharmacological relevance
Qiju Dihuang Pill (QJDHW) is a traditional Chinese medicine (TCM) formulation used for treating ocular diseases, and clinical evidence has demonstrated its efficacy in managing dry eye disease (DED). Age-related DED is characterized by tear film instability and ocular surface dysbiosis. Current therapeutic agents face challenges in achieving sustained efficacy. TCM offers a multi-targeted intervention strategy for age-related DED. However, the molecular mechanisms underlying QJDHW, a classical TCM formulation, remain to be systematically elucidated.
Aim of the study
This study aims to investigate the proteomic profile of age-related DED in clinical practice and further explore the specific mechanisms of QJDHW in treating age-related DED.
Methods
We established a tripartite research framework integrating clinical phenotyping, omics profiling, and herbal intervention. This study enrolled 30 participants at the First Affiliated Hospital of Hunan University of Chinese Medicine: 15 with age-related DED and 15 healthy controls. All subjects met established diagnostic, inclusion, and exclusion criteria. Morning fasting blood samples were collected, with serum separated by centrifugation for subsequent data-independent acquisition (DIA) proteomic analysis. Biomarkers and associated mechanisms were identified through bioinformatics analysis. An age-related DED rat model was established to investigate the mechanism of QJDHW (Beijing Tongrentang). Methodologies included: ocular surface assessment via tear film breakup time (BUT), Schirmer's test (SIT), and corneal fluorescein staining; spatial learning/memory evaluation using the Morris water maze (platform crossings and escape latency); histopathological examination of corneal and lacrimal gland tissues through hematoxylin-eosin (HE) staining; and multi-platform quantification of complement regulators (C3, C5, CFHR3, CD59) and inflammatory markers (IL-6) using immunohistochemistry, Western blotting, and RT-qPCR.
Results
Proteomic analysis revealed significant enrichment of the complement and coagulation cascades pathway, with key differential proteins including C5, CD59, and CFHR3, etc (P < 0.05). High-dose QJDHW (2.43 g/kg/day) significantly improved tear secretion, prolonged tear film stability, and repaired the morphology and function of the cornea and lacrimal gland. Mechanistically, QJDHW upregulated complement inhibitors (CFHR3, CD59) while downregulating complement activators (C3, C5) and IL-6 expression.
Conclusion
QJDHW exerts therapeutic effects through modulation of the complement activation-inhibition system, effectively suppressing C3/C5-mediated inflammatory cascades. This study provides molecular-level evidence supporting TCM's holistic regulation strategy in age-related ocular disorders.
{"title":"Qiju Dihuang Pill modulates complement system in age-related dry eye disease: Tripartite validation through clinical-omics-animal integration","authors":"Lihao Chen , Jun Peng , Genyan Qin , Dongdong Li , Xiaolei Yao , Qinghua Peng","doi":"10.1016/j.jep.2026.121190","DOIUrl":"10.1016/j.jep.2026.121190","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div>Qiju Dihuang Pill (QJDHW) is a traditional Chinese medicine (TCM) formulation used for treating ocular diseases, and clinical evidence has demonstrated its efficacy in managing dry eye disease (DED). Age-related DED is characterized by tear film instability and ocular surface dysbiosis. Current therapeutic agents face challenges in achieving sustained efficacy. TCM offers a multi-targeted intervention strategy for age-related DED. However, the molecular mechanisms underlying QJDHW, a classical TCM formulation, remain to be systematically elucidated.</div></div><div><h3>Aim of the study</h3><div>This study aims to investigate the proteomic profile of age-related DED in clinical practice and further explore the specific mechanisms of QJDHW in treating age-related DED.</div></div><div><h3>Methods</h3><div>We established a tripartite research framework integrating clinical phenotyping, omics profiling, and herbal intervention. This study enrolled 30 participants at the First Affiliated Hospital of Hunan University of Chinese Medicine: 15 with age-related DED and 15 healthy controls. All subjects met established diagnostic, inclusion, and exclusion criteria. Morning fasting blood samples were collected, with serum separated by centrifugation for subsequent data-independent acquisition (DIA) proteomic analysis. Biomarkers and associated mechanisms were identified through bioinformatics analysis. An age-related DED rat model was established to investigate the mechanism of QJDHW (Beijing Tongrentang). Methodologies included: ocular surface assessment via tear film breakup time (BUT), Schirmer's test (SIT), and corneal fluorescein staining; spatial learning/memory evaluation using the Morris water maze (platform crossings and escape latency); histopathological examination of corneal and lacrimal gland tissues through hematoxylin-eosin (HE) staining; and multi-platform quantification of complement regulators (C3, C5, CFHR3, CD59) and inflammatory markers (IL-6) using immunohistochemistry, Western blotting, and RT-qPCR.</div></div><div><h3>Results</h3><div>Proteomic analysis revealed significant enrichment of the complement and coagulation cascades pathway, with key differential proteins including C5, CD59, and CFHR3, etc (<em>P</em> < 0.05). High-dose QJDHW (2.43 g/kg/day) significantly improved tear secretion, prolonged tear film stability, and repaired the morphology and function of the cornea and lacrimal gland. Mechanistically, QJDHW upregulated complement inhibitors (CFHR3, CD59) while downregulating complement activators (C3, C5) and IL-6 expression.</div></div><div><h3>Conclusion</h3><div>QJDHW exerts therapeutic effects through modulation of the complement activation-inhibition system, effectively suppressing C3/C5-mediated inflammatory cascades. This study provides molecular-level evidence supporting TCM's holistic regulation strategy in age-related ocular disorders.</div></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121190"},"PeriodicalIF":5.4,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145952308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Acorus calamus L., popularly called vacha or sweet flag, is used in Ayurveda, Unani, Siddha, Chinese and other traditional medicines to treat a wide range of illnesses, including neurological, gastrointestinal, respiratory, metabolic, kidney, and liver disorders.
Aim of the study
This study aims to investigate the vasorelaxation potential of β-asarone (BA), a key biomarker of the A. calamus, as well as BA-enriched essential oil (ACRO), supercritical CO2 extract (ACRE), and BA-free fraction of A. calamus.
Materials and methods
The vasoreactivity of BA, ACRO, ACRE, and the BA-free fraction of A. calamus was evaluated in an ex vivo system with isolated superior mesenteric atrial rings, including elucidating the mode of action. In vitro toxicity in rat smooth muscle cells and in vivo oral toxicity in Swiss albino mice were evaluated to determine the safety profile of the test extract and the molecule.
Results
BA, ACRO and ACRE exhibited promising vasorelaxation in isolated rat mesenteric arteries (1–30 μg/ml), whereas the BA-free fraction exhibited a negligible vasorelaxation response. BA-induced concentration-dependent relaxation responses were studied in rat mesenteric arteries, and it was found to be highly sensitive in modulating calcium channel function in vascular smooth muscle cells, suggesting L-type VDCC as a major putative target in vasorelaxation response. Further, BA, ACRO and ACRE were evaluated for cytotoxicity in vascular smooth muscle cells. In vitro toxicity of BA suggested its safety up to 30 μM in the MTT assay. However, ACRO and ACRE showed cytotoxicity and inhibition of cell proliferation beyond 30 μg/ml when incubated for 12h. In continuation, the results of in vivo toxicity showed that ACRO and ACRE exhibited potential toxicity and produced complete mortality with ACRO and partial mortality with ACRE within the observational period of seven days, although morbidity and observational changes were evident in all the treatment groups when given from the range of 300 mg/kg up to 2000 mg/kg.
Conclusions
This data suggests a classic case of efficacy-toxicity paradox in A.calamus, suggesting further study to retain efficacy and enhance safety.
{"title":"β-Asarone-induced vasorelaxation in isolated rat mesenteric artery: An efficacy vs toxicity paradox of Acorus calamus","authors":"Shweta Parashar , Munmun Kumar Singh , Kavita Singh , Uma Shankar , Suyashi Mishra , Sudeep Tandon , Ram Swaroop Verma , Debabrata Chanda","doi":"10.1016/j.jep.2026.121187","DOIUrl":"10.1016/j.jep.2026.121187","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div><em>Acorus calamus</em> L., popularly called vacha or sweet flag, is used in Ayurveda, Unani, Siddha, Chinese and other traditional medicines to treat a wide range of illnesses, including neurological, gastrointestinal, respiratory, metabolic, kidney, and liver disorders.</div></div><div><h3>Aim of the study</h3><div>This study aims to investigate the vasorelaxation potential of <em>β</em>-asarone (BA), a key biomarker of the <em>A. calamus</em>, as well as BA-enriched essential oil (ACRO), supercritical CO<sub>2</sub> extract (ACRE), and BA-free fraction of <em>A. calamus</em>.</div></div><div><h3>Materials and methods</h3><div>The vasoreactivity of BA, ACRO, ACRE, and the BA-free fraction of <em>A. calamus</em> was evaluated in an <em>ex vivo</em> system with isolated superior mesenteric atrial rings, including elucidating the mode of action. In vitro toxicity in rat smooth muscle cells and <em>in vivo</em> oral toxicity in Swiss albino mice were evaluated to determine the safety profile of the test extract and the molecule.</div></div><div><h3>Results</h3><div>BA, ACRO and ACRE exhibited promising vasorelaxation in isolated rat mesenteric arteries (1–30 μg/ml), whereas the BA-free fraction exhibited a negligible vasorelaxation response. BA-induced concentration-dependent relaxation responses were studied in rat mesenteric arteries, and it was found to be highly sensitive in modulating calcium channel function in vascular smooth muscle cells, suggesting L-type VDCC as a major putative target in vasorelaxation response. Further, BA, ACRO and ACRE were evaluated for cytotoxicity in vascular smooth muscle cells. In vitro toxicity of BA suggested its safety up to 30 μM in the MTT assay. However, ACRO and ACRE showed cytotoxicity and inhibition of cell proliferation beyond 30 μg/ml when incubated for 12h. In continuation, the results of <em>in vivo</em> toxicity showed that ACRO and ACRE exhibited potential toxicity and produced complete mortality with ACRO and partial mortality with ACRE within the observational period of seven days, although morbidity and observational changes were evident in all the treatment groups when given from the range of 300 mg/kg up to 2000 mg/kg.</div></div><div><h3>Conclusions</h3><div>This data suggests a classic case of efficacy-toxicity paradox in <em>A.</em> <em>calamus</em>, suggesting further study to retain efficacy and enhance safety.</div></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121187"},"PeriodicalIF":5.4,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145948786","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-08DOI: 10.1016/j.jep.2025.121147
Zishu Zhang , Heyun Tao , Kexin Mao , Fanying Meng , Shanghong Jiang , Junyi Chen , Xinmiao He , Xuefei Tian
<div><h3>Ethnopharmacological relevance</h3><div>Patients with intermediate and advanced hepatocellular carcinoma (HCC) often derive limited benefit from systemic therapy and experience a substantial symptom burden with deterioration in quality of life. These challenges underscore the need for safe, effective adjunctive or alternative therapies. The traditional Chinese medicine concept of Central Qi shows conceptual contemporary with contemporary understanding of the gut-liver axis and microbiome biology. Chinese herbal medicines centered on tonifying the central and replenishing qi (Buzhong Yiqi) are considered adjunctive or alternative therapeutic options for alleviating symptoms, improving quality of life, and enhancing tolerance and adherence to standard treatments.</div></div><div><h3>Aim of this review</h3><div>To delineate links among the interrelations among Central Qi, the gut microbiota, and HCC, to synthesize evidence on how Buzhong Yiqi medicines modulate the gut microbiota, and to elucidate the proposed mechanistic bases for their potential adjunctive effects in HCC. This review aims to provide a biologically plausible framework to inform microbiome-based mechanistic research and clinical translation.</div></div><div><h3>Methods</h3><div>Databases including PubMed, Web of Science, Elsevier ScienceDirect, CNKI, and Google Scholar were searched using predefined terms related to hepatocellular carcinoma, gut microbiota, and the gut-liver axis, nutrient absorption, metabolic regulation, immune modulation, and Buzhong Yiqi medicines, as well as their representative herbs. In vivo, in vitro, and clinical studies published from 2000 to October 2025 were included.</div></div><div><h3>Results</h3><div>Dysbiosis disrupts the metabolic pathways of bile acid, short-chain fatty acid, and tryptophan metabolic pathways, while compromising the intestinal barrier. This disruption can worsen appetite regulation and energy imbalance, as well as weaken antitumor immunity. Buzhong Yiqi medicines have been reported to enrich beneficial taxa, reduce pathogens and pathobionts, and improve microbial metabolite profiles, restoration of barrier integrity, improvements in nutrient intake and energy homeostasis. These findings are heterogeneous and largely derived from non-HCC models, but collectively suggest potential support of metabolic and immune homeostasis, with possible influence on the tumor immune microenvironment. Early exploratory data also indicate a potential interaction with immune checkpoint inhibitors, although its clinical significance remains uncertain.</div></div><div><h3>Conclusion</h3><div>Central Qi deficiency provides a biologically plausible conceptual framework linking impaired digestion, disrupted energy metabolism, microbial dysbiosis, and reduced immune responsiveness in HCC. Modulation of the GM through Buzhong Yiqi medicines may offer supportive metabolic and immunological benefits, but current evidence is preliminary, based mainly
民族药理学相关性：中晚期肝细胞癌（HCC）患者通常从全身治疗中获得有限的益处，并且伴随着生活质量的恶化而经历实质性的症状负担。这些挑战强调需要安全、有效的辅助或替代疗法。传统中医的中气概念与当代对肝肠轴和微生物生物学的理解显示出概念上的时代性。以补中益气为中心的中草药被认为是缓解症状、改善生活质量、增强耐受性和坚持标准治疗的辅助或替代治疗方案。本文旨在探讨中气、肠道菌群和HCC之间的相互关系，综合补中益气药物调节肠道菌群的证据，并阐明其在HCC中潜在辅助作用的机制基础。本综述旨在提供一个生物学上合理的框架，为基于微生物组的机制研究和临床转化提供信息。方法：检索PubMed、Web of Science、Elsevier ScienceDirect、中国知网（CNKI）和谷歌Scholar等数据库，使用肝细胞癌、肠道菌群、肠肝轴、营养吸收、代谢调节、免疫调节、补中益气药物及其代表性中药等相关术语进行检索。包括从2000年到2025年10月发表的体内、体外和临床研究。结果：生态失调破坏了胆汁酸、短链脂肪酸和色氨酸的代谢途径，同时损害了肠道屏障。这种破坏会加重食欲调节和能量失衡，并削弱抗肿瘤免疫。据报道，补中益气药物可以丰富有益类群，减少病原体和病原菌，改善微生物代谢谱，恢复屏障完整性，改善营养摄入和能量稳态。这些发现是异质的，主要来自非hcc模型，但总体上表明代谢和免疫稳态的潜在支持，可能影响肿瘤免疫微环境。早期的探索性数据也表明与免疫检查点抑制剂的潜在相互作用，尽管其临床意义仍不确定。结论：中枢性气虚提供了一个生物学上合理的概念框架，与HCC中消化受损、能量代谢紊乱、微生物生态失调和免疫反应性降低有关。通过补中益气药物调节GM可能提供支持性代谢和免疫益处，但目前的证据是初步的，主要基于相关发现，需要谨慎解释。主要的不确定性仍然存在于因果关系、hcc特异性机制和临床效果的一致性方面。未来的研究应优先考虑标准化制剂、机制验证、生物标志物引导分层和严格设计的临床试验，以阐明这些建议途径的临床相关性，并促进全球对综合治疗的接受。
{"title":"The Central Qi theory in traditional Chinese medicine: Gut microbiota modulation as a strategic target for hepatocellular carcinoma therapy","authors":"Zishu Zhang , Heyun Tao , Kexin Mao , Fanying Meng , Shanghong Jiang , Junyi Chen , Xinmiao He , Xuefei Tian","doi":"10.1016/j.jep.2025.121147","DOIUrl":"10.1016/j.jep.2025.121147","url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><div>Patients with intermediate and advanced hepatocellular carcinoma (HCC) often derive limited benefit from systemic therapy and experience a substantial symptom burden with deterioration in quality of life. These challenges underscore the need for safe, effective adjunctive or alternative therapies. The traditional Chinese medicine concept of Central Qi shows conceptual contemporary with contemporary understanding of the gut-liver axis and microbiome biology. Chinese herbal medicines centered on tonifying the central and replenishing qi (Buzhong Yiqi) are considered adjunctive or alternative therapeutic options for alleviating symptoms, improving quality of life, and enhancing tolerance and adherence to standard treatments.</div></div><div><h3>Aim of this review</h3><div>To delineate links among the interrelations among Central Qi, the gut microbiota, and HCC, to synthesize evidence on how Buzhong Yiqi medicines modulate the gut microbiota, and to elucidate the proposed mechanistic bases for their potential adjunctive effects in HCC. This review aims to provide a biologically plausible framework to inform microbiome-based mechanistic research and clinical translation.</div></div><div><h3>Methods</h3><div>Databases including PubMed, Web of Science, Elsevier ScienceDirect, CNKI, and Google Scholar were searched using predefined terms related to hepatocellular carcinoma, gut microbiota, and the gut-liver axis, nutrient absorption, metabolic regulation, immune modulation, and Buzhong Yiqi medicines, as well as their representative herbs. In vivo, in vitro, and clinical studies published from 2000 to October 2025 were included.</div></div><div><h3>Results</h3><div>Dysbiosis disrupts the metabolic pathways of bile acid, short-chain fatty acid, and tryptophan metabolic pathways, while compromising the intestinal barrier. This disruption can worsen appetite regulation and energy imbalance, as well as weaken antitumor immunity. Buzhong Yiqi medicines have been reported to enrich beneficial taxa, reduce pathogens and pathobionts, and improve microbial metabolite profiles, restoration of barrier integrity, improvements in nutrient intake and energy homeostasis. These findings are heterogeneous and largely derived from non-HCC models, but collectively suggest potential support of metabolic and immune homeostasis, with possible influence on the tumor immune microenvironment. Early exploratory data also indicate a potential interaction with immune checkpoint inhibitors, although its clinical significance remains uncertain.</div></div><div><h3>Conclusion</h3><div>Central Qi deficiency provides a biologically plausible conceptual framework linking impaired digestion, disrupted energy metabolism, microbial dysbiosis, and reduced immune responsiveness in HCC. Modulation of the GM through Buzhong Yiqi medicines may offer supportive metabolic and immunological benefits, but current evidence is preliminary, based mainly ","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"360 ","pages":"Article 121147"},"PeriodicalIF":5.4,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145948820","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-08DOI: 10.1016/j.jep.2026.121184
Ting Huang , Yang Yang , Jianyan Yan , Lining Yu , Xiaoyan Mao , Lifen Chen , Li Xu , Junfang Zhang , Yijie Sun , Liming Chong , Jia Zeng
Ethnopharmacological relevance
Zhibo Qingliang (ZBQL) formula, a clinically validated traditional Chinese medicine (TCM) comprising sixteen herbs, has been used for over two decades to treat chronic glomerulonephritis (CGN), particularly in cases presenting with proteinuria due to damp-heat retention and declining healthy energy. Despite its extensive clinical application, the preclinical safety profile of the modernized ZBQL granule remained unestablished.
Aim of the study
This study aimed to evaluate the safety of ZBQL dry extract through single- and repeated-dose oral toxicity studies in Sprague Dawley (SD) rats, including the determination of the maximum tolerated dose (MTD) and no-observed-adverse-effect level (NOAEL).
Materials and methods
A single-dose study was conducted by the administration of 30.0 g/kg ZBQL extract (43.3 × human dose) to SD rats, followed by 14-day observation. A 13-week repeated-dose study with a 4-week recovery period was performed at daily doses of 6.0, 12.0, and 15.0 g/kg (equivalent to 8.7, 17.3, and 21.6 × human dose), with an administration volume of 30 mL/kg. Parameters included clinical signs, body weight, food consumption, hematology, coagulation, blood biochemistry, urinalysis, organ weights (absolute and relative), and histopathology.
Results
The single-dose MTD was ≥30.0 g/kg, with only transient diarrhea observed. In the repeated-dose study, the NOAEL was 12.0 g/kg. Dose-dependent reductions in body weight and food intake were observed in both male and female rats, attributed to the high viscosity. Reversible, non-adverse changes included mild hematological shifts, biochemical alterations consistent with reduced nutritional intake, and adaptive histopathological findings such as centrilobular hepatocyte hypertrophy and alveolar macrophage aggregation at the high dose. All changes resolved following the recovery period.
Conclusions
ZBQL extract demonstrated a wide safety margin in rats, with no evidence of direct organ toxicity. Observed effects were reversible and consistent with exaggerated pharmacological activity or adaptive responses. These findings support the continued clinical development of ZBQL granule as a safe and effective TCM-based therapeutic.
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