Pharmaceutical Biology最新文献

Context: Physalis peruviana L. (Solanaceae), also known as Poha, has been used in traditional medicine since pre-Columbian times, particularly in treating cancer.

Objective: To study the chemical composition and potential medicinal properties of Poha.

Materials and methods: The fresh fruits and aerial parts of Poha were extracted. The isolation of extract yields a novel withanolide (physaperuvin K; 1) from the edible fruit, and seven withanolides (2-8), including a rare chlorinated withanolide (physalolactone; 2) from the aerial parts. Structure elucidation/determination was performed, some acetate derivatives were prepared (2a-6a), and the compounds were evaluated with in vitro assays indicative of anti-inflammatory activity.

Results: The structure of 1 was elucidated through NMR spectroscopic analyses. The absolute configuration of compound 2 was determined using single-crystal X-ray diffraction. Compounds 1, 2, and 3 exhibited inhibition of tumor necrosis factor-α-induced nuclear factor-kappa B (NF-κB) activity with IC₅₀ values of 10, 60, and 40 nM, respectively, without causing cytotoxicity at a concentration of 50 μM. Furthermore, compounds 1-3 reduced nitric oxide (NO) production in lipopolysaccharide-activated RAW 264.7 mouse macrophage cells with IC₅₀ values ranging from 0.32 to 13.3 μM without overt cytotoxicity. Overall, acetylation did not significantly impact activity, except for compound 4, wherein the IC₅₀ values in the NF-κB and NO assays were reduced from 11.0 to 0.33 μM, and 1.8 to 0.24 μM, respectively.

Conclusions: These findings enhance our understanding of Poha's constituents and potential medicinal properties. One of the most bioactive compounds identified in this study, physaperuvin K, is found in edible fruit.

Context: WDJ-S4, a standardized herbal formula, has been prescribed for refractory functional dyspepsia (FD) in Korea, but the detailed mechanisms are lacking.

Objective: The present study investigates the acceleration of gastrointestinal (GI) motility by WDJ-S4 and its potential mechanisms.

Materials and methods: For five days, WDJ-S4 (50, 100 and 200 mg/kg) or mosapride (3 mg/kg) was orally given to BALB/c mice. After 20 h of fasting, loperamide (10 mg/kg, i.p.) was given to the mice except for normal group. To assess gastric emptying or intestinal propulsion, 500 μL of 0.05% phenol red or 200 μL of 5% charcoal diet was given once orally.

Results: Loperamide delayed gastric emptying and intestinal propulsion, while WDJ-S4 ameliorated peristaltic dysfunction, evidenced by reductions of remaining phenol red in the stomach and a marked increase of charcoal propulsion in the intestine. WDJ-S4 also normalized levels of acetylcholine and acetylcholine-related enzymes, including choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), in the gastric antrum and jejunum. C-kit level and smooth muscle contraction-related genes were elevated by WDJ-S4 in both the gastric antrum and jejunum.

Conclusion: Overall, WDJ-S4 can effectively promote GI motility. The efficacy is associated with modulation of acetylcholine pathway and the interstitial cells of Cajal (ICCs) activation. All the results provide scientific evidence supporting the clinical usage of WDJ-S4 for FD.

Context: Vascular inflammation is a key process in the pathogenesis of atherosclerosis, which is regulated by NF-κB pathway. Yiqi Wenyang decoction (YQWY), a Traditional Chinese medicine (TCM) formula, has anti-inflammatory properties and may inhibit this pathway, potentially offering anti-atherosclerotic effects.

Objective: The purpose of this study is to investigate the effects of YQWY on atherosclerosis and the underlying mechanism. Materials and methods: ApoE^-/- mice were fed a Western diet and administered with YQWY (low or high dose), atorvastatin, or vehicle for 13 weeks. The size of atherosclerotic plaques was assessed using ORO staining. Vascular inflammation was evaluated with IF or IHC staining. The mechanisms and signaling pathways underlying the effect of YQWY on vasculature were studied using transcriptomic analysis and were validated in vitro in endothelial cells and macrophages.

Results: YQWY attenuated atherosclerotic plaque development which was associated with reduced vascular inflammation as demonstrated by transcriptomic analysis of aorta. This was verified by reduced expression of proinflammatory chemokines, adhesion molecules, and inflammatory cytokines in aortas from YQWY-treated mice at both mRNA and protein levels. Mechanistically, YQWY suppressed NF-κB activation in endothelial cells and, to a lesser extent, macrophages possibly.

Discussion and conclusions: YQWY protects against vascular inflammation and atherosclerosis by suppressing NF-κB pathway, suggesting the potential of YQWY and its active ingredients as novel anti-atherosclerotic therapeutics.

Context: Xiao-Luo-Wan (XLW), a classical prescription in traditional Chinese medicine, has therapeutic effects on uterine fibroids (UFs). Herein, its anti-UF effects were examined using a systematic pharmacological method.

Objective: To explore the active ingredients of XLW via mass spectrometry and its potential effects on UFs by network pharmacology, molecular docking, and experimental validation.

Materials and methods: A mass spectrometer was used to scrutinize the composition of the XLW drug-containing serum. The critical targets and potential mechanisms of XLW against UFs were predicted by network pharmacology and molecular docking. Next, human uterine leiomyoma cells (UMCs) were treated with 20%, 30%, or 40% XLW serum for 24 h, 48 h or 72 h. Cell viability was analyzed via a CCK-8 assay, and cell apoptosis and the cell cycle were examined via flow cytometry. The predicted targets were further identified by RT-PCR and western blotting.

Results: There were 16 chemical components identified in XLW drug-containing serum, with 53 target genes predicated in the treatment of UFs. The molecular binding of core targets, including TRIM9, NF-κB and p38MAPK, was relatively stable to components, especially buergerinin B, cedrol and ent-15B-16-epoxy- kauan-17-ol. The in vitro experiments revealed that the IC₅₀ of XLW in UMCs was 63.21%, and the anti-UF effects of XLW may be closely associated with targets that inhibit cell proliferation and promote cell apoptosis by regulating TRIM9, NF-κB and p38MAPK expression.

Discussion and conclusions: The integration of mass spectrometry, network pharmacology, molecular docking and biological experiments revealed the key constituents of XLW and its pharmacological mechanism in UFs, which may help in the discovery of therapeutic agents for treating UFs.

Context: Flavones, the key active components in Traditional Chinese Medicine (TCM), have demonstrated antidepressant activity. Given the numerous animal studies conducted, a systematic analysis is essential to provide a valuable reference for future research.

Object: This study investigated the antidepressant activity of flavones based on animal models and summarized the underlying mechanisms.

Methods: We systematically searched 7 bibliographic Databases as of August 12, 2023, such as Web of Science, PubMed, China National Knowledge Infrastructure, etc. The meta-analysis was performed using either the random or fixed-effect model, supplemented by trial sequential analysis (TSA). The Grading of Recommendations, Assessment, Development and Evaluations (GRADE) approach was used to assess the quality of evidence.

Results: A total of 25 studies involving 458 mice were included, identifying five flavones (baicalin, baicalein, apigenin, luteolin, vitexin) with antidepressant activity. Compared to the control group, flavones significantly reduced immobility time in forced swimming and tail suspension tests. Flavones also decreased serum and hippocampal levels of interleukin (IL)-1β and tumor necrosis factor-alpha (TNF-α), reduced nuclear factor kappa B (NF-κB) levels, and increased brain-derived neurotrophic factor (BDNF) levels. Relative to the positive group, flavones raised IL-6, sucrose preference rate, and corticosterone (CORT) levels, with no significant differences in other factors. The TSA showed the efficacy of flavones for treating depression with adequate 'information size' for the primary outcome.

Conclusions: The results demonstrate that flavones exert protective effects against depression in mice, primarily by stimulating neurotrophic factors and modulating inflammatory pathways. These findings emphasize their potential as promising candidates for the development of novel antidepressant therapies.

Context: Myocardial fibrosis is a common manifestation of end-stage cardiovascular disease, but there is a lack of means to reverse fibrosis. Astragaloside IV (AS-IV), the major active component of Astragalus membranaceus Fisch. ex Bunge Fabaceae, possesses diverse biological activities that have beneficial effects against cardiovascular disease.

Objective: This systematic review aims to summarize the anti-fibrosis effect of AS-IV in animal models (rats or mice only) and its underlying mechanisms, and provide potential directions for the clinical use of AS-IV.

Methods: PubMed, EMBASE, Web of Science, CNKI, Wanfang database, and SinoMed were searched from inception to 31 December 2024. The following characteristics of the included studies were extracted and summarized: animal model, route of administration, dose/concentration, measurement indicators, and potential mechanisms. The quality of the included studies was assessed used a 10-item scale from SYRCLE.

Results and conclusion: AS-IV represents a promising multi-target candidate for myocardial fibrosis treatment in the 24 eligible studies included in the analysis. This systematic review is the first to comprehensively evaluate the anti-fibrosis mechanisms of AS-IV across heterogeneous cardiovascular disease animal models, including myocardial infarction, hypertension, ischemia-reperfusion injury, and myocarditis. The underlying mechanisms of the anti-fibrosis effects of AS-IV may include collagen metabolism, anti-apoptosis, anti-inflammation and, pyroptosis, antioxidants, improving mitochondrial function, regulating senescence, etc. Current evidence remains preclinical, with critical gaps in toxicological profiles, human safety thresholds, and clinical adverse reaction data. Future research must integrate robust toxicological evaluations, optimized combination therapies, and adaptive clinical trials to validate translational potential.

Context: Aging leads to senile osteoporosis (SOP), marked by bone loss and increased fracture risk. Macrophages, as active immune cells in bone tissue, play an important role in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) during aging. Wen-Shen-Tong-Luo-Zhi-Tong Decoction (WSTLZTD), a traditional Chinese herbal formula, has been clinically validated for its efficacy in treating SOP. However, the specific mechanisms by which WSTLZTD exerts its anti-SOP effects-particularly through modulating macrophage senescence-remain unclear.

Objective: The study aims to elucidate the role of WSTLZTD in macrophage senescence and SOP.

Materials and methods: Aged mice received low, medium, high-dose WSTLZTD. Bone loss was evaluated via micro-computed tomography, hematoxylin and eosin staining and osteocalcin, tartrate-resistant acid phosphatase marker analysis. Macrophage senescence detection (β-galactosidase staining, p16, p21) and molecular mechanisms by Western blot, immunohistochemistry, immunofluorescence method were investigated. Macrophage-conditioned medium's effects on BMSC osteogenesis and mitochondrial function were assessed through alkaline phosphatase, Alizarin Red S staining, reactive oxygen species and JC-1 mitochondrial membrane potential (ΔΨm) assays.

Results: In vivo experiments demonstrated that WSTLZTD effectively ameliorated macrophage senescence and osteoporosis in naturally aged mice. Mechanistically, high-dose WSTLZTD attenuated senescence in bone marrow-derived macrophages by mediating LONP1, concurrently suppressing the cyclic GMP-AMP synthase (cGAS)/STING signaling pathway in BMSCs, thereby enhancing osteogenic differentiation of BMSCs. In vitro studies further confirmed that WSTLZTD-containing serum attenuated the senescent phenotype of macrophages. Notably, the LONP1 inhibitor, LONP1-IN-2, was found to diminish the anti-senescence effects of WSTLZTD on macrophages and BMSC osteogenesis.

Discussion and conclusion: WSTLZTD potentially modulate macrophage senescence via LONP1, which subsequently suppresses the activation of the cGAS/STING pathway in BMSCs, ultimately promoting their osteogenic differentiation and ameliorating osteoporosis.

Context: Cancer therapy remains a challenge in healthcare, particularly in the context of triple-negative breast cancer (TNBC), where targeted therapies are still scarce.

Objective: Addressing this issue, our study explores a novel targeting approach using small extracellular vesicles (sEVs) isolated from cow milk, functionalized with hyaluronic acid (HA) to target the overexpressed cluster of differentiation 44 (CD44) cell surface receptor in TNBC cells.

Materials & methods: A method for isolating sEVs from cow milk was optimized, and the obtained sEVs were fully characterized in terms of size, morphology, and protein markers. Subsequently, milk-derived sEVs were covalently bound with HA of varying molecular weights (MW, 20-60 kDa, 250 kDa, 1000-1600 kDa) and binding and internalization dynamics were investigated. Breast cancer cell lines, MDA-MB-231 (TNBC and CD44+) and MCF-7 (CD44-), were used as in vitro models to evaluate CD44 selectivity.

Results: The binding and internalization studies unveiled enhanced selectivity of functionalized sEVs for CD44-overexpressing cells compared to non-functionalized sEVs. Notably, higher MW HA exhibited enhanced binding capacity, with partial internalization occurring through CD44 endocytic mechanisms.

Discussion and conclusion: In summary, this work introduces a sEVs isolation method and sheds light on the role of HA MW in enhancing cellular uptake of CD44 overexpressing cancer cells.

Context: As a naturally occurring terpenoid found in Cannabis sativa L. (Cannabaceae), cannabidiol (CBD) has gained public and industry interest for the purposes of personal well-being as a foodstuff and pharmaceutical. Despite a number of publications on CBD toxicology, many have significant limitations, especially those relating to genotoxicity. These include poor characterization of the CBD extract and/or lack rigor in conforming to accepted regulatory guidelines and best practice. A number of regulatory agencies have highlighted these issues and requested additional genotoxicity data to help ensure the safe use of CBD.

Objective: To provide insights into the genotoxicity of a CBD isolate and its lipid carrier.

Materials and methods: We have conducted an in vitro mammalian cell micronucleus (OECD 487) and a bacterial reverse mutagenicity assay (Ames test) (OECD 471) in a CBD isolate (97% > CBD) with its carrier.

Results: The samples tested were non-mutagenic, as determined in the Ames test. The in vitro micronucleus assay conducted was negative for genotoxicity, with no statistically significant increases in the incidences of micronucleated cells observed at any dose compared to negative controls.

Conclusions: These studies confirm that this CBD rich isolate in combination with its carrier, are unlikely to post any genotoxic hazard at exposure levels expected in foods.

Context: Gansui [Euphorbia kansui T. N. Liou ex S.B.Ho (Euphorbiaceae)] has been reported to inhibit the proliferation of non-small cell lung cancer (NSCLC) cells; however, its underlying pharmacological mechanism remains unclear.

Objective: To investigate the potential effects and mechanisms of Gansui in blocking NSCLC progression.

Materials and methods: The targets of Gansui's components and NSCLC-related targets were obtained through public database and published studies. Functional enrichment analysis was performed using the clusterProfiler R package. STRING database was used for protein-protein interaction analysis. CytoHubba plugin was applied to get the hub genes. Molecular docking was applied to assess the binding affinities between the hub targets and the crucial components. Kidjolanin was used to treat A549 and NCI-H1385, and its effects on cell viability, sensitivity of paclitaxel and expression levels of hub genes were investigated by cell counting kit-8 assay, flow cytometry and qPCR.

Results: A total of 16 Gansui active ingredients and 337 targets were collected, of which 298 targets overlapped with NSCLC-related genes. STAT3, EGFR, GRB2, AKT2, AKT3 and PIK3CA were identified as hub genes. The components in Gansui, including kidjoranin 3-O-β-digitoxopyranoside, cynotophylloside B, 13-Oxyingenol-dodecanoate, and kidjolanin had good binding affinity with the hub targets. Kidjolanin inhibited the viability of NSCLC cells, promoted apoptosis and inhibited the expression of hub genes. Kidjolanin also enhanced the proliferation inhibition and apoptosis of NSCLC cells induced by paclitaxel.

Discussion and conclusion: Gansui exerts anti-NSCLC effects via multiple downstream targets, implying its potential in NSCLC treatment.