摘要
目的采用网络药理学的方法预测四物汤活性成分的潜在美白作用,并通过体外实验进行功效验证。方法通过TCMSP和PubChem数据库筛选四物汤的活性成分及其靶点,结合Gene Cards数据库获取黑色素合成相关靶点。利用Cytoscape 3.9.1软件绘制“四物汤-活性成分-潜在靶点”关系图并筛选关键成分,随后通过STRING数据库构建蛋白质相互作用网络。采用DAVID数据库进行GO和KEGG分析。通过酪氨酸酶抑制实验、DPPH自由基清除实验和羟自由基清除实验对关键成分进行功效验证。结果通过网络药理学分析筛选出四物汤活性成分71个,与黑色素合成相关的交集靶点74个,其中核心靶点7个,KEGG富集通路主要为PI3K-AKT信号通路。在酪氨酸酶抑制实验中,β-谷甾醇和蔗糖对酪氨酸酶的平均抑制率达98%;芍药内酯苷在质量浓度为0.6 mg/mL和1.0 mg/mL时对酪氨酸酶的抑制率低于35%,而在其余质量浓度下对酪氨酸酶的抑制率高于50%;四物汤对酪氨酸酶无抑制作用。在DPPH自由基清除实验中,β-谷甾醇、蔗糖、芍药内酯苷和四物汤在质量浓度为0.2 mg/mL时对DPPH自由基的清除率均低于18%;β-谷甾醇在质量浓度为1.0 mg/mL时对DPPH自由基的清除率为96%;芍药内酯苷在质量浓度为0.6 mg/mL和0.8 mg/mL时对DPPH自由基的清除率为4.94%和11.64%;四物汤对DPPH自由基的抑制能力随着浓度升高而增强,清除率达59%。在羟自由基清除实验中,β-谷甾醇对羟自由基的最大清除率为52%;蔗糖和芍药内酯苷对羟自由基的最大清除率为88%,但芍药内酯苷在质量浓度为0.2 mg/mL时对羟自由基无清除作用;四物汤在实验浓度范围内对羟自由基的最大清除率仅为33.24%。结论基于网络药理学和体外实验证实,四物汤通过调控黑色素合成通路发挥美白作用,其活性成分β-谷甾醇、蔗糖和芍药内酯苷均表现出酪氨酸酶抑制活性和抗氧化能力,这为开发四物汤作为天然美白剂提供了科学依据。
Objective To predict the potential whitening effect of the active ingredients in Siwu Decoction using the method of network pharmacology and verify the efficacy through in vitro experiments.Methods TCMSP and PubChem databases were used to screen the active ingredients and their targets of Siwu Decoction,and combined with Gene Cards database to obtain melanin synthesis related targets.Cytoscape 3.9.3 software was used to draw a relationship diagram of"Siwu Tang-active ingredients-potential targets"and screen for key components.STRING database was used to construct a protein interaction network.DAVID database was used to perform GO and KEGG analysis.The efficacy of key components was validated through tyrosinase inhibition experiments,DPPH radical scavenging experiments,and hydroxyl radical scavenging experiments.Results Through network pharmacology analysis,71 active components of Siwu Decoction were screened out.There were 74 intersecting targets related to melanin synthesis,among which there were 7 core targets.The main KEGG enrichment pathway was the PI3K-AKT signaling pathway.In the tyrosinase inhibition experiment,the average inhibition rates ofβ-sitosterol and sucrose on tyrosinase reached 98%.The inhibitory rate of albiflorin on tyrosinase was less than 35%at concentrations of 0.6 mg/mL and 1.0 mg/mL,while the inhibitory rate on tyrosinase was higher than 50%at other concentrations.Siwu Decoction had no inhibitory effect on tyrosinase.In the DPPH radical scavenging experiment,the scavenging rates ofβ-sitosterol,sucrose,albiflorin and Siwu Decoction on DPPH radicals were all below 18%at a concentration of 0.2 mg/mL.The scavenging rate ofβ-sitosterol on DPPH free radicals was 96%at a concentration of 1.0 mg/mL.The scavenging rates of albiflorin on DPPH free radicals were 4.94%and 11.64%at concentrations of 0.6 mg/mL and 0.8 mg/mL.The inhibitory ability of Siwu Decoction on DPPH free radicals increased with increasing concentration,with a clearance rate of 59%.In the hydroxyl radical scavenging experiment,the clearance rate ofβ-sitosterol on hydroxyl radicals reached 52%.Sucrose and albiflorin had a hydroxyl radical scavenging rate of 88%,but albiflorin had no hydroxyl radical scavenging effect at a concentration of 0.2 mg/mL.The maximum scavenging rate of Siwu Decoction on hydroxyl radicals within the experimental concentration range was only 33.24%.Conclusion Based on network pharmacology and in vitro experiments,it has been confirmed that Siwu Decoction exerts whitening effects by regulating the melanin synthesis pathway.Its active ingredients,β-sitosterol,sucrose,and albiflorin,all exhibit tyrosinase inhibitory activity and antioxidant capacity,providing a scientific basis for the development of Siwu Decoction as a natural whitening agent.
作者
张慧
区海彤
王婴
王岩
ZHANG Hui;OU Haitong;WANG Ying;WANG Yan(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;Teaching Experiment Center,Guangdong Pharmaceutical University,Guangzhou 510006,China)
出处
《广东药科大学学报》
2025年第6期127-136,共10页
Journal of Guangdong Pharmaceutical University
基金
广东药科大学2024年大学生创新创业训练计划(S202410573030)
广东药科大学2025年大学生创新创业训练计划(20250430210)。
关键词
网络药理学
四物汤
美白作用
Network pharmacology
Siwu Decoction
whitening
