Phytomedicine最新文献

Ischemic stroke ranks as the second leading cause of global mortality and disability. Although reperfusion is crucial for salvaging brain tissue, it carries the risk of secondary injuries, such as ferroptosis. Gastrodin, a neuroprotective compound found in Chinese herbal medicine, may regulate this process. However, its impact on stroke-induced ferroptosis remains unclear.

Objective: This research endeavors to probe Gastrodin's influence on post-ischemic ferroptosis, deciphering its mechanisms and assessing its therapeutic promise.

Methods: We developed rat models of middle cerebral artery occlusion/reperfusion (MCAO/R) and created oxygen-glucose deprivation/reoxygenation (OGD/R)-damaged PC12 cell models. Gastrodin was administered to assess ferroptosis using Prussian blue staining and fluorescence probes. To investigate the effects of gastrodin on the xCT/GPX4 and ACSL4/LPCAT3 pathways, we employed molecular docking, immunofluorescence, Western blotting, and quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, we used transmission electron microscopy and JC-1 fluorescence probes to examine mitochondrial integrity and function.

Results: Our study demonstrated that gastrodin significantly reduced iron accumulation and lipid peroxidation in the brains of MCAO/R rats and OGD/R-injured PC12 cells. It suppressed reactive oxygen species (ROS) and ameliorated mitochondrial membrane potential. It potentiates the xCT/GPX4 axis while repressing the ACSL4/LPCAT3 pathway, leading to improved mitochondrial architecture and function, notably characterized by decreased mitochondrial membrane potential, reduced ROS levels, and increased formation of mitochondrial cristae. By modulating the xCT/GPX4 and ACSL4/LPCAT3 pathways, gastrodin mitigated ferroptosis in ischemic stroke, thereby preserving mitochondrial structural and functional integrity. This study provides novel mechanistic insights into gastrodin's therapeutic potential for treating ischemic stroke, highlighting the importance of traditional Chinese medicine in modern medical therapy.

Background: Andrographolide is a medicinal compound which possesses anti-SARS-CoV-2 activity. A number of cellular targets of andrographolide have been identified by target predictions and computational studies.

Purpose: However, a potential cellular target of andrographolide has never been explored in SARS-CoV-2 infected lung epithelial cells. We aimed to identify cellular pathways involved in andrographolide-mediated anti-SARS-CoV-2 activity.

Methods: The viral infection was determined by immunofluorescence staining, enzyme-linked immunosorbent assay and focus-forming assay. Proteomic analysis was employed to identify cellular pathways and key proteins controlled by andrographolide in the human lung epithelial cells Calu-3 infected by SARS-CoV-2. Immunofluorescence staining was used to test protein expression and localization. Western blot and realtime PCR were utilized to elucidate gene expression. Cellular glutathione level was examined by a reduced/oxidized glutathione assay. An ectopic gene expression was delivered by plasmid transfection.

Results: Gene ontology analysis indicates that proteins involved in nuclear factor erythroid 2-related factor 2 (NRF2)-regulated pathways were differentially expressed by andrographolide. Notably, andrographolide increased expression and nuclear localization of the transcription factor NRF2. In addition, transcriptional expression of GCLC and glutamate-cysteine ligase modifier subunit (GCLM), which are NRF2 target genes, were induced by andrographolide. We further find that infection of SARS-CoV-2 resulted in a reduction of glutathione level in Calu-3; the effect that was rescued by andrographolide. Moreover, andrographolide also induced expression of the glutathione producing enzyme GCLC in SARS-CoV-2 infected lung epithelial cells. Importantly, an ectopic over-expression of GCLC or treatment of N-acetyl-L-cysteine in Calu-3 cells led to a decrease in SARS-CoV-2 infection.

Conclusion: Collectively, our findings suggest the interplay between GCLC-mediated glutathione biogenesis induced by andrographolide and the anti-SARS-CoV-2 activity. The glutathione biogenesis and recycling pathways should be further exploited as a targeted therapy against SARS-CoV-2 infection.

Background: Owing to high sensitivity and ability for absolute quantification, the droplet digital polymerase chain reaction (ddPCR) is widely used for viral and bacterial detection. However, few studies have been conducted on the application of ddPCR to identify the original plant species used in traditional Chinese medicine and Chinese patent medicine.

Purpose: In this study, we investigated the feasibility of using ddPCR to differentiate between Notopterygium incisum and N. franchetii to establish a sensitive and quantitative method for quality control of herbal materials and preparations.

Methods: Specific minor groove binding (MGB) probes and primers were designed based on stable single nucleotide polymorphisms. The ddPCR experimental conditions were designed and optimised according to the results of multiplex PCR and qPCR, which ultimately confirmed the limits of detection and quantification (LOD and LOQ, respectively) of the method for Notopterygii Rhizoma et Radix. Additionally, the original plant species of Notopterygii Rhizoma et Radix in Jiuwei Qianghuo pills circulating in the market were identified.

Results: The results of the multiplex PCR and qPCR indicated that the probes and primers were specific. Furthermore, a Qsep analyser and Sanger sequencing were used to confirm that the specific amplification products of N. incisum and N. franchetii were 283 and 206 bp, respectively. The optimised ddPCR system was employed to determine the LOD to be 0.000816 ng/µl, and LOQ of N. incisum and N. franchetii to be 0.00408 and 0.003312 ng/µl, respectively. In addition, Notopterygii Rhizoma et Radix in four Jiuwei Qianghuo pills was amplified and successfully identified using ddPCR assays.

Conclusion: This study established a multiplex ddPCR method using MGB probes to identify Notopterygii Rhizoma et Radix, providing a foundation for the identification and quantification of multi-source Chinese herbal medicines.

Background: Artemisia spp. have been used for millennia in traditional medicine to treat a variety of ailments, including malaria. Extracts of Artemisia afra and A. annua remain widely used throughout Africa for healthcare purposes, notably to prevent and/or treat malaria. However, the modes of action of these plant extracts remain unclear, with contradictory reports regarding the presence and role of artemisinin in both plants.

Purpose: The aim of this study was to identify differences in the antimalarial mode of action of A. afra and A. annua by measuring their phenolic profiles and comparing their effect on parasite metabolism in vitro.

Methods: In this work, we analyzed the phenolic profile of A. afra and A. annua extracts through high-performance liquid chromatography (HPLC), detected and quantified artemisinin through HPLC and mass spectrometry (MS), and performed comparative HPLC-MS metabolomic analysis on in vitro-cultured Plasmodium falciparum trophozoites to elucidate the potential modes of action of these plant extracts.

Results: A. afra contained only trace amounts of artemisinin and elicited a different parasite metabolic response compared to A. annua, which contained significantly more artemisinin and correlated closely with the parasite response profile elicited by purified artemisinin. A. annua impacted parasite glutathione metabolism in agreement with the established redox activity of artemisinin, while A. afra had an effect on lipid precursors.

Conclusions: This study reveals that A. afra and A. annua have divergent effects on Plasmodium falciparum metabolism and provides support for ongoing efforts exploring the use of A. afra for the treatment of malaria.

Background: Promoting the coupling of osteogenesis and angiogenesis is a crucial strategy for the treatment of postmenopausal osteoporosis (PMOP). Estrogen deficiency induces ferroptosis, which is closely associated with the pathophysiology of PMOP. Sarsasapogenin (SAR) is a natural sapogenin with anti-oxidative effects. However, it is unclear whether SAR has a protective role against the impaired osteogenesis and angiogenesis coupling in PMOP. In this study, we evaluated the efficacy of SAR in estrogen deficiency-induced osteoporosis and explored the underlying mechanisms.

Methods: Bone marrow mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) were utilized to assess the in vitro effects of SAR on the coupling of osteogenesis and angiogenesis. In vivo experiments involved bilateral ovariectomy (OVX)-induced osteoporosis in mice and glutathione peroxidase 4 (GPX4)-knockout (KO) mice. Mice were orally administered SAR (5 or 10 mg/kg/d) for a duration of 12 weeks. The direct target of SAR was investigated through molecular docking, a cellular thermal shift assay, and surface plasmon resonance. Additionally, RNA sequencing was employed to elucidate the underlying mechanisms.

Results: SAR treatment improved cell viability and osteogenic differentiation while inhibiting ferroptosis in iron dextran-induced BMSCs. Furthermore, SAR enhanced the production of slit guidance ligand 3 (SLIT3) in these cells, which stimulated angiogenesis by activating its receptor, roundabout human homolog 1 (ROBO1), in HUVECs. An in vitro model of ferroptosis induced by erastin demonstrated that SAR promoted the coupling of osteogenesis and angiogenesis by upregulating the BMSCs-SLIT3/HUVECs-ROBO1 axis. Activation of GPX4 was identified as a contributing factor to the effects of SAR on this coupling. Transfection of GPX4 small interfering RNA (siRNA) in BMSCs negated the impact of SAR on the BMSCs-SLIT3/HUVECs-ROBO1 axis. Additionally, SAR was found to directly interact with GPX4, enhancing protein stability, with an equilibrium dissociation constant of 44.6 μM. Notably, SAR did not increase SLIT3, ROBO1, or indicators of osteogenesis or angiogenesis in GPX4-KO mice.

Conclusions: These findings underscore the significance of restoring the GPX4/SLIT3/ROBO1 axis in promoting the coupling of angiogenesis and osteogenesis. SAR mitigates PMOP, in part, by activating the BMSCs-SLIT3/HUVECs-ROBO1 axis, with GPX4 serving as an upstream signaling modulator responsible for SLIT3 production. Our observations provide experimental evidence supporting the clinical application of SAR in the treatment of PMOP.

Background: Oral squamous cell carcinoma (OSCC) is one of the most common malignancies. However, there is no effective treatment for OSCC.

Purpose: This study aimed to identify a natural compound with significant efficacy against OSCC and elucidate its primary mechanism of action.

Methods: An FDA-approved drug library and an MCE autophagy-related molecular compound library were screened through high-throughput screening to identify an effective natural compound against OSCC. The IC50 value of sanguinarine (Sang) in OSCC cells was determined using a CCK8 assay. Immunoblotting and immunofluorescence staining were used to assess the effect of Sang on autophagic flux in OSCC cells. Changes in the acidic lysosomal environment were evaluated using RFP-GFP-LC3B and LysoSensor Green DND-189. Furthermore, limited proteolysis-coupled mass spectrometry (LiP-MS) and virtual screening techniques were utilized to identify direct binding targets of Sang, which were subsequently validated by surface plasmon resonance (SPR) and microscale thermophoresis (MST). Molecular docking combined with molecular dynamics analysis identified the binding site between the target protein and Sang. In vitro and in vivo investigations with mutant plasmids confirmed this finding.

Results: Screening led to the identification of the naturally occurring autophagy modulator Sang as a potent inhibitor of OSCC progression. Moreover, Sang impaired lysosomal function through reducing lysosomal-associated membrane proteins, inhibiting lysosomal proteolysis, and altering the lysosomal pH. These effects contributed to defects in autophagic clearance and subsequently suppressed OSCC progression. Notably, Sang bound the phenylalanine 26 (F26) residue in pyruvate kinase M2 (PKM2) and inhibited PKM2 enzymatic activity, subsequently suppressing transcription factor EB (TFEB) expression to inhibit lysosomal function and blocking autophagic flux in OSCC cells.

Conclusion: Our results demonstrate for the first time that Sang can suppress the PKM2/TFEB axis, and influence lysosomal function, thereby blocking autophagy and inhibiting the progression of OSCC, making it a promising therapeutic option for the treatment of OSCC.

Background: Heart failure (HF) represents an advanced stage of various cardiovascular disorders, with its elevated admission rates and resultant health economic burden posing an ongoing global concern.

Purpose: To evaluate the health and economic benefits of herbal medicine (HM) for patients with HF.

Study design: Population-based cohort study.

Methods: A five-year retrospective cohort study was carried out at a nationally recognized hospital in China. The study utilized propensity score matching (PSM) to match patients with HF. Chi-square tests were used to analyze dichotomous variables, and t-tests were employed for continuous variables. Logistic regression was used to examine hospital readmission rates, while multiple linear regression was utilized to evaluate direct medical costs. Statistical significance was set at p < 0.05.

Results: After implementing PSM, 1924 HF patients were included in the analysis. The study identified two significant risk factors affecting the readmission rates: age over 65 years (adjusted odds ratio (OR) = 1.25, 95 % confidence interval (CI) [1.02, 1.53]) and smoking (adjusted OR = 1.31, 95 % CI [1.01, 1.70]). Additionally, patients who received adjunctive HM treatment exhibited a significantly lower readmission rate compared to those without HM treatment (adjusted OR = 0.76, 95 % CI [0.64, 0.92]). Furthermore, the use of HM during patient hospitalization did not significantly impact direct medical expenses but instead provided positive health economic benefits (incremental cost-effectiveness ratio (ICER) = 98.52). Factors influencing direct routine medical costs included over 65 years of age (Coef = 60.78, 95 % CI [36.25, 85.31]), and cardiac function classification (New York Heart Association (NYHA) III: Coef = 1979.92, 95 % CI [1401.82, 2558.03]; NYHA IV: Coef = 6052.48, 95 % CI [5166.59, 6938.38]).

Conclusions: The integration of HM in patients with HF reduced readmission rates without a notable increase in direct medical costs, and the expense of HM remains an economically range indicating positive health economic outcomes.

Background: Inflammation is the body's innate reaction to foreign pathogens and serves as a self-regulating mechanism. However, the immune system can mistakenly target the body's own tissues, triggering unnecessary inflammation. For millennia, medicinal plants have been employed for the treatment of diseases. One such plant, Rhaphidophora peepla, has demonstrated potential anti-inflammatory properties. However, the precise mechanism underlying its anti-inflammatory effects remains elusive.

Study design: For this study, validation of target molecules by different experimental approaches and employing two different in vivo experiments were tried to improve the immunopharmacological value of Rhaphidophora peepla.

Purpose: Our goal is to elucidate the mechanism through which the ethanol extract of Rhaphidophora peepla (Rp-EE) demonstrates anti-inflammatory properties, both in vivo and in vitro.

Method: Rp-EE was phytochemically analyzed with gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). Bioinformatic analysis with protein-protein interaction (PPI) networks and Kyoto Encyclopedia of Genes and Genomes (KEGG), nitric oxide (NO) assay, MTT assay, RT-PCR, ELISA, luciferase assay, CETSA, hematoxylin and eosin (H&E) staining, and Western blotting analysis were used to evaluate anti-inflammatory activity of Rp-EE and its mechanism.

Results: Rp-EE significantly reduced inflammatory responses including nitric oxide (NO) release induced by lipopolysaccharide (LPS) at the non-cytotoxic concentrations in vitro, and HCl/EtOH-induced gastritis and LPS-induced acute lung injury models in vivo. Mechanistically, it was revealed that Rp-EE can specifically target spleen tyrosine kinase (Syk) and transforming growth factor β-activated kinase 1 (TAK1) to suppress the phosphorylation levels of nuclear factor (NF)-κB subunits (p65 and p50) and activator protein (AP)-1 subunits (c-Jun and c-Fos).

Conclusion: Rp-EE can inhibit inflammatory reactions managed by Syk and TAK1, resulting in suppressing the Syk/AKT/NF-κB and TAK1/MAPK/AP-1 signaling pathways. These findings lead us to a possibility that Rp-EE can be developed as a promising anti-gastric ulcer and anti-lung injury remedy.