Skin aging commonly manifests as deepening wrinkles, loss of elasticity, and weakened barrier function, resulting from the long-term accumulation of multiple biological processes. Dermal fibroblasts, as the primary source of extracellular matrix, not only provide structural support but also play an active role in aging. On one hand, they undergo intrinsic aging due to telomere shortening, mitochondrial decline, and dysregulation of signaling pathways (e.g., TGF-β, mTOR). On the other hand, they release inflammatory cytokines and proteases via the senescence-associated secretory pattern (SASP), disrupting keratinocyte function, melanin distribution, immune surveillance, and even microvascular and adipose tissue functions. This destabilizes the matrix equilibrium and exacerbates inflammation, creating a vicious cycle. While strategies like dasatinib/quercetin, rapamycin, or retinol show promise, they remain constrained by transdermal efficiency and targeting limitations. This review aims to elucidate these mechanisms and interactions, providing insights for developing more effective anti-aging interventions.

Non-alcoholic fatty liver disease (NAFLD) represents a chronic liver disorder with widespread global prevalence, primarily attributed to hepatic lipid accumulation, oxidative stress, and inflammatory responses. In the realm of traditional Chinese medicine, Ganoderma lucidum is predominantly utilized for its hepatoprotective properties. The objective of this study was to isolate and identify novel bioactive compounds capable of mitigating hepatic fat accumulation. An in vitro steatosis model was established using oleic acid-induced HepG2 cells to evaluate the total triglyceride (TG) content across various components. Fractionation of the compounds was guided by the observed reduction in TG content, employing multiple chromatographic techniques to successfully isolate ten Ganoderma triterpenes. Structural elucidation was achieved through 1D and 2D NMR spectroscopy, supplemented by additional spectroscopic methods. This investigation led to the identification of two previously unreported lanostane-type triterpenes (1–2) alongside eight known analogues (3–10). Compound 1 and 2 exhibit structural distinctions from the other compounds, primarily in the substituents at positions C-3, C-7, and C-15, as well as in the spatial orientation of these substituents. In vitro experiments were conducted to assess the efficacy of various compounds in inhibiting lipid accumulation. Compound 1-5 demonstrated a significant reduction in TG levels within the OA-induced HepG2 cell model (p < 0.05). In comparison to the model group, Compound 1 demonstrated a moderate lipid-lowering effect, (2.11 mmol/gprot vs. 2.70 mmol/gprot, p < 0.003). Conversely, Compound 2 exhibited a significantly more pronounced lipid-lowering effect, (1.27 mmol/gprot vs. 2.70 mmol/gprot, p < 0.0001). Furthermore, when compared with the positive control drug, the lipid-lowering efficacy of Compound 2 was significantly superior to that of Compound 1. Furthermore, the application of network pharmacology, molecular docking, and molecular dynamics simulations elucidated the mechanism of action underlying the effects of methyl ganoderenic acid A(2).

OBJECTIVE:This study aims to investigate how hyperoside (HYP) alleviates oxidative stress-induced osteoporosis, its molecular mechanisms, and its impact on osteoblast differentiation, oxidative damage, and the estrogen-PI3K/VEGF signaling pathway. METHODS:The osteoblast differentiation model was induced using dexamethasone, and osteoblast-related markers like ALP, NO, GSH, MDA, and SOD were measured post-HYP intervention. A zebrafish model was used to assess HYP's impact on ROS and bone formation. Network pharmacology identified key oxidative stress and osteoporosis targets, with HYP's binding affinity confirmed via molecular docking and simulation. RT-qPCR verified the expression of key pathway targets. RESULTS:HYP can counteract dexamethasone-induced inhibition of osteoblast differentiation, boost ALP, NO, GSH, and SOD levels, and lower MDA levels in osteoblasts. It also reduces ROS accumulation and enhances bone formation in zebrafish. Network pharmacology identified a common oxidative stress and osteoporosis target, with molecular docking confirming HYP's stable binding. RT-qPCR showed HYP significantly upregulates SRC, PI3K, AKT1, and e-NOS, activating the estrogen-PI3K/VEGF pathway. CONCLUSION:HYP plays an anti-oxidative stress effect through targeted regulation of estrogen-PI3K/VEGF signal axis, and then promotes osteoblast differentiation and bone formation, which provides a new potential candidate drug and experimental basis for the treatment of osteoporosis.

To screen for anti-PMO active components in Cornus officinalis based on ERβ enzyme activity using affinity ultrafiltration and molecular docking techniques.ERβ enzyme and UHPLC-Q-Exactive Orbitrap MS were employed to analyze the anti-PMO activity and predict the components of different concentrations of ethanol extracts of C. officinalis. Molecular docking was used to verify the interaction mechanism between small molecule ligands and ERβ. The anti-PMO activity of the predicted components was further verified using MC3T3-L1 cells. The results showed that the ERβ enzyme can serve as a target enzyme for screening anti-PMO components. The 50% ethanol extract exhibited the best activity. UHPLC-Q-Exactive Orbitrap MS identified and analyzed 17 potential active components. Based on the binding rate and molecular docking results, the following three active components were identified: sweroside, 4-hydroxycinnamic acid, and cornuside. In vitro activity validation confirmed that these components have potential anti-PMO effects. The ERβ enzyme can be used as a potential target enzyme for screening anti-PMO active components in traditional Chinese medicine. The combination of affinity ultrafiltration and molecular docking provides an effective and rapid method for active component screening, offering valuable references for the targeted therapy, mechanism exploration, and quality control of traditional Chinese medicine.

BACKGROUND:Verbascoside, a compound classified as a phenylethanol glycoside in Dihuang, has been the subject of modern pharmacological investigations. These studies have revealed its noteworthy antioxidant, anti-inflammatory, memory-enhancing, neuroprotective, antitumor, and various other pharmacological properties. While verbascoside exhibits favorable antioxidant effects, its precise mechanism of action in ameliorating osteoporosis through the treatment of oxidative stress remains unclear. METHODS:This study employed CCK8, ALP, ELISA, and ROS staining techniques to examine the osteoporotic effects of verbascoside on zebrafish and MC3T3-E1 cells. Additionally, this study aimed to investigate the molecular mechanism by which verbascoside improves osteoporosis by mitigating oxidative stress. To identify the common targets of verbascoside in relation to oxidative stress and osteoporosis, network pharmacology and molecular dynamics simulation were employed. The construction of the verbascoside - oxidative stress - osteoporosis - potential target gene network aimed to identify the core targets, while the mechanism of action was elucidated through KEGG analysis, and the accuracy was confirmed by assessing the mRNA expression of the targets. RESULTS:In vivo experiments demonstrated that verbascoside exhibited therapeutic effects on osteoporosis and reduced ROS production in zebrafish. In vitro experiments further revealed that verbascoside enhanced the proliferation and differentiation of MC3T3-E1 cells, thereby improving the oxidative stress status of osteoblasts. Thirteen core targets and estrogen signaling pathways were identified through the application of network pharmacology. The pivotal role of the estrogen signaling pathway in facilitating the ability of verbascoside to mitigate oxidative stressinduced osteoporosis was substantiated by the modulation of target protein mRNA expression. CONCLUSION:The findings underscore the considerable therapeutic potential of verbascoside in ameliorating osteoporosis through the alleviation of oxidative stress, thus establishing it as a promising compound for the treatment of this condition.

Depression is a psychological disorder with significant global impact. It is widely hypothesized that this disorder is associated with neuroinflammation, which disrupts neural homeostasis through various pathways. This study aims to investigate the effective compounds and mechanisms of Black Ginseng under forest (BG) in combating neuroinflammation. Utilizing methods such as UPLC-QE Orbitrap-MS, network pharmacology, molecular docking, and cell biology, the efficacy of BG was demonstrated, and its active components were identified. Cell viability and apoptosis were assessed using Trans well migration assays and flow cytometry. The mRNA expression of target genes was confirmed through real-time quantitative PCR (RT-qPCR), elucidating the anti-neuroinflammatory mechanism. The results indicated that BG exhibited a more pronounced effect on ameliorating neuroinflammatory conditions compared to Ginseng under forest (FG). The main active components were found through research and development, including Ginsenoside F1, Ginsenoside Rk1, Ginsenoside Rg3, etc. Among these, Ginsenoside F1 emerged as the most potent active component for treating neuroinflammation, as evidenced by reduced cell migration and apoptosis. The study demonstrates that BG can modulate the PI3K-Akt signaling pathway, leading to a reduction in the expression levels of AKT1, MAPK1, PIK3CA, EGFR, and other mRNAs. These findings suggest that BG is a promising natural antidepressant, providing both theoretical and experimental foundations for the development of new antidepressants based on BG and its active components.

INTRODUCTION:Postmenopausal osteoporosis, a skeletal disorder induced by estrogen deficiency, is commonly addressed in clinical practice through the use of Cuscutae semen and its compound formulations, owing to their significant therapeutic efficacy. OBJECTIVE:This study sought to systematically identify bioactive compounds present in Cuscutae semen that interact with estrogen receptor β (ESR2) using Affinity Ultrafiltration combined with Ultra-Performance Liquid Chromatography-Quadrupole-Orbitrap Mass Spectrometry (UPLC-QE-Orbitrap-MS) to find possible therapy options for PMOP. METHODS:The Cuscutae semen extract was fractionated using a C18 column to obtain fractions based on distinct polarity. Preliminary evaluation of anti-osteoporotic activity was conducted in MC3T3-E1 cells by assessing osteoprotegerin (OPG) expression. Affinity Ultrafiltration integrated with UPLC-QE-Orbitrap-MS was utilized to screen for ligands binding to ESR2, followed by molecular docking to validate the interaction mechanisms. The osteogenic effects of the identified compounds were further confirmed through CCK-8 proliferation assays, OPG quantification, and alkaline phosphatase (ALP) activity analysis. RESULTS:The fraction of Cuscutae semen extract eluted with 100% methanol exhibited significant anti-osteoporotic activity. Three flavonoids-Astragalin, Isorhamnetin, and Quercitrin-that interact with ESR2 were successfully identified. In vitro validation demonstrated the efficacy of these compounds. CONCLUSION:This study presents a comprehensive strategy that integrates Affinity Ultrafiltration, UPLC-QE-Orbitrap-MS, and bioactivity validation to efficiently identify ESR2-targeted postmenopausal osteoporosis compounds in Cuscutae Semen. The findings offer both theoretical and empirical foundations for the development of innovative therapeutics for postmenopausal osteoporosis.

The primary etiology of postmenopausal osteoporosis is estrogen deficiency. The enzyme aromatase (CYP19A1) serves as the key rate-limiting enzyme in the conversion of androgens to estrogens. In this study, ginseng was selected as the subject of investigation. The study identified that the 50 % ethanol extract of ginseng exhibited the most potent activity in the determination of CYP19A1 enzyme activity. This extract was subsequently analyzed using UHPLC-QE Orbitrap-MS in conjunction with mass spectrometry molecular network technology. To identify the active components effective against postmenopausal osteoporosis, enzyme ultrafiltration affinity, molecular docking, and kinetic simulation techniques were employed. Cross-analysis of binding energy and affinity rate results revealed that ginsenoside Re and ginsenoside Rf possessed the highest absolute binding energy and affinity values, establishing them as the most effective active components. The mechanisms involving ALP, OPG, apoptosis, and qPCR were validated in vitro to confirm the anti-PMOP effects of these active ingredients. This study offers an efficient and rapid method for screening natural products to identify active components for the treatment of postmenopausal osteoporosis.

Rhizoma Drynariae (RD), derived from the desiccated rhizome of Drynaria fortunei (Kunze) J. Sm., has been researched for its potential therapeutic efficacy in modulating inflammatory responses (IR) and treating osteoporosis (OP). Nevertheless, the molecular mechanisms through which RD attenuates OP by modulating the IR microenvironment have yet to be thoroughly elucidated. Consequently, this study aimed to assess the impact of RD on IR and OP through integrated zebrafish and cellular model assays, focusing on the quantification of neutrophil recruitment, bone mineralization area, and the levels of tumor necrosis factor-α, nitric oxide, interleukin-6, and alkaline phosphatase. The experimental data demonstrate that the < 3 kDa RD fraction (RDE-2) effectively mitigates IR and OP models across both zebrafish and cellular models. Utilizing a novel dose-effect weighted network pharmacology, we identified kaempferol-3-O-rutinoside, procyanidin B2, and prunin as the primary bioactive constituents of RDE-2. Combined with untargeted metabolomics analysis, potential targets for their action were identified. Subsequently, a network comprising active ingredients, core targets, metabolic targets, and metabolites was constructed to elucidate their functional interactions. RT-qPCR analysis revealed a significant upregulation in the mRNA expression levels of AKT1, ESR1, ESR2, MMP9, PI3K, SRC, EP300, NCOA3, Runx2, and CREBBP. The findings suggest that RD has the potential to mitigate OP by modulating the IR microenvironment through the ER/PI3K-EP300 signaling axis. Through the integration of dose-effect weighted network pharmacology and metabolomic analysis, our study advances beyond existing descriptive research on RD and pioneers the elucidation of the ER/PI3K-EP300 axis, thereby offering a novel mechanistic explanation.

Ginseng exhibits pharmacological efficacy in the treatment of female postmenopausal osteoporosis (PMOP), although its specific active constituents remain unidentified. This study aims to identify the anti-PMOP active components of ginseng by screening against the ESR2 enzyme. Using osteoprotegerin (OPG) as a biomarker, the most effective ESR2-active components of ginseng were determined through experiments with MC3T3-E1 cells. The potential active components within ginseng were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole-exactive orbitrap mass spectrometry (UHPLC-QE Orbitrap-MS) and further validated through molecular docking and molecular dynamics simulations. The single-target affinity ultrafiltration method was employed to assess the affinity rate (AR%) of the ESR2 enzyme by analyzing peak areas, leading to the identification of key active components of ginseng effective against PMOP through molecular docking analysis. Experimental findings indicated that ginseng extract exhibits compatibility with the ESR2 enzyme, with the 95 % ethanol extract demonstrating the highest activity. A total of 35 potential active components were identified through UHPLC-QE Orbitrap-MS analysis. Following a comprehensive evaluation of molecular docking binding energy and AR% results, Ginsenoside Rf, Ginsenoside Re, and Ginsenoside Rb1 were confirmed as the principal active components of ginseng effective against PMOP. The survival rate, OPG content, and alkaline phosphatase (ALP) activity index, a marker of osteogenic differentiation, were utilized to confirm the final key active components. In this study, the ESR2 enzyme was identified as the active target, and a combination of cell experiments, molecular docking, kinetic simulations, and affinity ultrafiltration was employed to effectively screen the active anti-PMOP constituents of ginseng.

BACKGROUND:Dioscoreae Rhizoma (DR) is a plant recognized for its dual medicinal and edible applications, exhibiting notable therapeutic efficacy, particularly in the treatment of postmenopausal osteoporosis (PMOP) associated with estrogen deficiency. OBJECTIVE:This study sought to systematically identify bioactive compounds present in DR that interact with estrogen receptor β (ESR2) and employ affinity ultrafiltration in conjunction with UPLC-QE-Orbitrap-MS to find possible therapy options for PMOP. METHODS:In this study, a C18 column was employed to fractionate the DR extract into distinct fractions, and the optimal active site in DR was identified based on its osteoprotegerin (OPG) content in MC3T3-E1 cells. To identify the DR components exhibiting high binding affinity for ESR2, affinity ultrafiltration coupled with UPLC-QE-Orbitrap-MS was utilized. These findings were further corroborated through molecular docking and molecular dynamics simulations. To further validate the osteogenic effects of the identified compounds, CCK-8 proliferation assays, along with OPG and alkaline phosphatase (ALP) activity assays, were employed. RESULTS:The 30% DR fraction demonstrated significant anti-PMOP activity. Acacetin, Adenosine, and Procyanidin B2 are recognized as the principal active constituents responsible for the anti-PMOP effects of DR. CONCLUSION:This study introduces a comprehensive approach combining affinity ultrafiltration with UPLC-QE-Orbitrap-MS and molecular docking to efficiently identify ESR2-targeted therapeutic compounds for PMOP in DR. The findings offer both theoretical and empirical foundations for the advancement of novel therapeutic strategies for PMOP.

Adenosine, a nucleoside, regulates various systems, such as the cardiovascular, immune, and nervous systems, by binding to Adenosine receptors. To elucidate the role of Adenosine in Osteoporosis, this study employed an experimental approach involving the stimulation of bone formation in osteoporotic zebrafish and the reduction of reactive oxygen levels in a glucocorticoid-induced zebrafish model of Osteoporosis with oxidative stress. Adenosine significantly promoted the proliferation of MC3T3-E1 cells and increased the activity of alkaline phosphatase (ALP). It also elevated nitric oxide, glutathione, and superoxide dismutase levels while decreasing malondialdehyde. The target of Adenosine’s impact on Osteoporosis with oxidative stress was elucidated through network pharmacology, revealing the PI3K/Akt pathway, a finding corroborated through RT-qPCR analysis. Hence, we present the mechanism through which Adenosine can be utilized to prevent and manage Osteoporosis accompanied by oxidative stress, distinct from the previously identified mode of action of Adenosine in osteoporosis treatment.

In order to explore the research and development of oyster in the field of medicine and food homology,response surface methodology was used to optimize the preparation process of oyster protease-depeptidase,and its effects on testosterone secretion and oxidative stress in mouse leydig cells were studied.Based on the investigation of hydrolysis degree as the evaluation index,a biomimetic enzymatic hydrolysis method was employed to optimize the preparation process of oyster protein enzymolysis peptides using response surface analysis,building upon the foundation of single-factor experiments.Simultaneously,a hydrogen peroxide(H2O2)-induced oxidative damage model was established using mouse testicular interstitial cells(TM3),and the effects of oyster protein enzymolysis peptides on testosterone(T)secretion and oxidative stress were investigated through assessments of cell viability,DAPI staining,testosterone secretion level,superoxide dismutase(SOD)activity,and malondialdehyde(MDA)content in TM3 cells.The results showed that the optimal enzymatic hydrolysis conditions for oyster protein enzymolysis peptides were as follows:Substrate-to-solvent ratio of 1∶10 g/mL,gastric protease concentration of 1.1％,hydrolysis time of 1.0 h,pancreatic protease concentration of 2.1％,and hydrolysis time of 3.1 h.Under these conditions,the degree of hydrolysis was determined to be 39.43％±0.42％.Oyster protein enzymolysis peptides exhibited varying degrees of proliferative activity on H2O2-induced TM3 cells,significantly(P＜0.05)increasing testosterone secretion,SOD enzyme activity,and reducing MDA levels in TM3 cells.The most pronounced effects were observed at a concentration of 200 μg/mL of oyster protein enzymolysis peptides.In conclusion,the optimization of enzymatic hydrolysis process using response surface methodology proved to be effective and feasible.Oyster protein enzymolysis peptides was found to extremely significant promote TM3 cell proliferation,increase testosterone secretion,enhance SOD enzyme activity,and reduce MDA levels(P＜0.01).

Purpose:Zhixiao Tang (ZXT), a traditional Chinese compound prescription, has been used clinically to treat pneumonia in China. However, the underlying mechanism of ZXT treatment in pneumonia is still unclear. The present study aimed to reveal the potential mechanism of ZXT in pneumonia using a strategy combining metabolomics and network pharmacology. Methods:Initially, the chemical compositions were identified by UPLC-QE-Orbitrap-MS, while the prediction of potential signal pathways was performed through network pharmacology. To assess the anti-inflammatory properties of ZXT in the context of pneumonia, models of 16HBE cells induced by LPS and zebrafish induced by CuSO4 were established to measure levels of inflammatory markers and apoptosis. Subsequently, the differential changes of endogenous metabolites in cells caused by ZXT were examined using metabolomics technology, and the molecular docking analysis of key targets was carried out using Autodock Vina software. Ultimately, the validation of the primary pathways and targets was conducted through quantitative RT-PCR and Western blot techniques. Results:A total of 75 compounds were identified through UPLC-QE-Orbitrap-MS analyses. Network pharmacological analysis shows that it plays an anti-inflammatory role in C-type lectin receptor signaling pathway. After ZXT intervention, the inflammatory factors and apoptosis in cells were significantly reduced. Metabonomics analysis showed that 18 metabolites changed significantly. Four key genes were identified, which exhibited partial compatibility with the findings of network pharmacology. Molecular docking analysis confirmed the substantial affinity of the primary targets for ZXT. Furthermore, ZXT exerted a suppressive effect on neutrophil migration, down-regulated the expression of pro-inflammatory cytokine genes, and inhibited the up-regulation of the Dectin-1/SYK/NF-κB signaling pathway. In vivo cell experiments also yielded consistent experimental outcomes. Conclusion:This study enhances comprehension of the pharmacological mechanism underlying ZXT's efficacy in pneumonia treatment, thereby establishing a scholarly basis for future research and clinical utilization of ZXT in pneumonia management.

Introduction: The industrial processing of corn (Zeamays L.) generates by-products such as corn silk, straw peels, and straw core, which contribute to adverse environmental impacts. Our study aimed to investigate sustainable approaches for mitigating these effects by evaluating the hypoglycemic potential and mechanisms of ethyl acetate fractions derived from these corn derivatives.Methods: We employed glucose consumption assays, high glucose stress tests, UPLC-QE-Orbitrap-MS analysis, molecular docking, and simulations to assess their components and efficacy. Antioxidant capacities were evaluated using DPPH, FRAP, ABTS, and •OH scavenging assays.Results: Notably, the ethyl acetate fraction extracted from straw peels (SPE) exhibited a high concentration of flavonoids and phenolic compounds along with pronounced hypoglycemic activity and antioxidant capacity. SPE significantly enhanced glucose consumption in insulin-resistant HepG2 cells while protecting HUVECs against damage caused by high glucose levels. Molecular docking analyses confirmed the interaction between active compounds and α-glucosidase as well as α-amylase, while molecular dynamic simulations indicated stability at their binding sites.Discussion: In conclusion, the hypoglycemic and antioxidative properties observed in corn by-products such as straw peels, corn silk, and straw core can be attributed to the inhibition of α-glucosidase and α-amylase activities, coupled with their rich phenolic and flavonoid content. These findings highlight the potential of these by-products for applications in healthcare management and their sustainable utilization, demonstrating significant value in the use of agricultural residues.

BACKGROUND:Osteoclasts are integral to the advancement of osteoporosis (OP), and their generation under conditions of oxidative stress (OS) involves various pathways. However, the specific mechanism through which the natural antioxidant kaempferol (KAE) mitigates the influence of OS on osteoclasts remains somewhat uncertain. This study aims to evaluate the effect of KAE on osteoclast formation under OS and explore its possible mechanism. METHODS:Zebrafish were used to observe the effects of KAE on OP and OS. OP and OS "double disease targets" network pharmacology were used to predict the action target and mechanism of KAE on OP under OS. The effects of KAE on osteoclast differentiation induced by OS were evaluated using RWA264.7 cells induced by LPS. To elucidate the potential mechanism, we detected the expression of related factors and target genes during induction. RESULTS:The presence of KAE exhibited potential in improving the conditions of OP and OS in zebrafish. KAE can reduce the OS of RAW 264.7 cells stimulated by LPS, inhibit the formation of osteoclasts, and change the level of related factors of OS, and reduce the increase of TRAP. The utilization of network pharmacology and target gene expression assay revealed that KAE exerted a down-regulatory effect on the expression of proto-oncogene tyrosine protein kinase (SRC), nuclear factor kappa-B (NF-κB), Serine/Threonine Kinase-1 (AKT1), Nitric Oxide Synthase 3 (NOS3) and Matrix Metallopeptidase-2 (MMP2). CONCLUSION:Based on the results of this study, KAE may effectively mitigate OS and impede the formation of osteoclasts through the SRC/NF-κB-AKT/NOS3 axis.

The purpose of this study was to conduct a screening and comparative analysis of proteolytic peptides with anti-inflammatory properties derived from three different ginseng concoctions:Sundried ginseng,red ginseng and black ginseng.Ginseng proteins were extracted from three different types of ginseng products using a low-temperature leaching method.Subsequently,the extracted proteins underwent enzymatic digestion using alkaline protease,neutral protease,and pepsin through a stepwise enzyme digestion method.This process yielded three distinct enzyme digestion products,namely BGP(black ginseng proteolytic peptide),RGP(red ginseng proteolytic peptide),and SGP(sundried ginseng proteolytic peptide).The samples were subjected to separation using ultrafiltration membranes,resulting in the acquisition of ultrafiltration fractions with distinct molecular weights.Subsequently,the ultrafiltration fractions were further separated utilizing ultrafiltration membranes to obtain fractions with varying molecular weights.The fraction exhibiting the most potent anti-inflammatory activity was determined through the application of a lipopolysaccharide(LPS)-induced RAW264.7 inflammation model.The impact of the active fractions on the secretion of nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin-1β((IL-1β),and interleukin-6(IL-6)by RAW264.7 cells was assessed using enzyme immunoassay.The amino acid composition and content of the three proteolytic peptides were examined.Multivariate statistical analysis was employed to identify the distinct amino acids in the three ginseng concoctions and investigate their correlation with the inhibition of cytokine secretion by RAW264.7 cells.The findings of the study indicated that the proteolytic peptide fraction with a molecular weight of less than 1 kDa in the three ginseng products exhibited the most pronounced impact on the proliferation of RAW264.7 cells compared to the other fractions.Additionally,this fraction significantly suppressed the secretion of NO,TNF-α,IL-6,and IL-1β at concentrations ranging from 50～200 μg/mL(P＜0.05).Notably,at a concentration of 200 iig/mL,the three groups receiving proteolytic peptide administration demonstrated the most potent inhibitory effect on cytokine release.Furthermore,the inhibitory effect of BGP-4 on cytokine release surpassed that of RGP-4 and SGP-4,exhibiting a statistically significant difference(P＜0.05).All three peptides consisted of 17 amino acids,however,their compositions exhibited significant variations.Notably,phenylalanine exhibited the highest content,and the differential amino acids present in the three ginseng concoctions were closely associated with the inhibition of inflammatory factor secretion.This study represents an initial exploration into the impact of concoctions on the anti-inflammatory properties of ginseng,identifying distinct amino acids among different concoctions.These findings offer a valuable reference for the formulation of ginseng concoctions.

Testicular dysfunction (TDF) is characterized by testosterone deficiency and is caused by oxidative stress injury in Leydig cells. A natural fatty amide named N-benzylhexadecanamide (NBH), derived from cruciferous maca, has been shown to promote testosterone production. Our study aims to reveal the anti-TDF effect of NBH and explore its potential mechanism in vitro. This study examined the effects of H2O2 on cell viability and testosterone levels in mouse Leydig cells (TM3) under oxidative stress. In addition, cell metabolomics analysis based on UPLC-Q-Exactive-MS/MS showed that NBH was mainly involved in arginine biosynthesis, aminoacyl-tRNA biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, the TCA cycle and other metabolic pathways by affecting 23 differential metabolites, including arginine and phenylalanine. Furthermore, we also performed network pharmacological analysis to observe the key protein targets in NBH treatment. The results showed that its role was to up-regulate ALOX5, down-regulate CYP1A2, and play a role in promoting testicular activity by participating in the steroid hormone biosynthesis pathway. In summary, our study not only provides new insights into the biochemical mechanisms of natural compounds in the treatment of TDF, but also provides a research strategy that integrates cell metabolomics and network pharmacology in order to promote the screening of new drugs for the treatment of TDF.

目的:研究山羊豆抗非小细胞肺癌的药效物质基础和潜在作用机制.方法:利用非小细胞肺癌A549细胞,给予山羊豆90％乙醇提取物及不同极性萃取物,比较各部位对A549细胞的增殖抑制率,筛选潜在活性部位,通过细胞划痕实验、Tran-swell 细胞迁移实验研究山羊豆活性部位对细胞迁移活力的影响.运用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用(UPLC-Q-Exactive-MS/MS)技术对筛选得到的部位的化学成分进行分析鉴定,利用网络药理学方法筛选抗非小细胞肺癌的核心靶点,在DAVID数据库进行GO和KEGG富集分析,在微生信平台上进行可视化分析.使用Cytoscape软件建立成分-核心靶点-通路网络图,运用Autodock vina软件进行分子对接并应用Pymol软件作可视化处理,分析、预测作用机制.结果:山羊豆乙酸乙酯部位能够有效抑制A549细胞增殖和迁移,山羊豆碱、槲皮苷、阿福豆苷等15个成分为山羊豆抗非小细胞肺癌的潜在药效成分,网络药理学及分子对接结果表明,黄酮类成分与EGFR、HRAS、MAPK等靶点对接吻合度较好.结论:山羊豆抵抗非小细胞肺癌可能是通过多成分、多靶点起作用,该研究可为山羊豆抗肺癌功能的开发应用及质量评价提供科学依据.