【摘要】 目的：基于网络药理学、分子对接和动物实验对参苓白术散缓解顺铂所致肾毒性（cisplatin nephrotoxicity, C-IN）的核心活性成分及潜在作用机制进行探讨。方法：运用中药系统药理学数据库与分析平台等数据库及文献检索初步获取参苓白术散组方中药的活性成分并预测其靶点；利用在线人类孟德尔遗传数据库、人类基因数据库、Drugbank数据库获取C-IN的靶点；取参苓白术散活性成分靶点与C-IN疾病相关靶点录入Venny 2.1.0软件，绘制韦恩图，得到两者的交集靶点。利用STRING数据库构建交集靶点的蛋白质相互作用（protein-protein interaction, PPI）网络并进行拓扑分析筛选核心靶点；运用DAVID数据分析平台进行基因本体（gene ontology, GO）富集分析及京都基因与基因组百科全书（kyoto encyclopedia of genes and genomes, KEGG）信号通路富集分析；选取关键靶点及核心活性成分，使用AutoDock软件进行分子对接。SPF级雄性C57BL/6J小鼠24只随机分为对照组、模型组、减毒组及参苓白术散组，每组6只，模型组和减毒组小鼠每3天腹腔注射1次顺铂注射液(3 mg·kg^-1)诱导肾损伤，减毒组同时灌胃给予参苓白术散治疗(3 g·kg^-1),参苓白术散组仅灌胃给予参苓白术散(3 g·kg^-1),对照组每天给予等量的生理盐水。30天后处死小鼠，采用电感耦合等离子体质谱检测肾组织内铂离子含量；脲酶法测定血清尿素氮（blood urea nitrogen, BUN）水平；肌氨酸氧化酶法测定血清肌酐（creatinine, Cre）的水平；HE染色观察肾组织病理变化；PCR检测肾组织AKT1 mRNA、TP53 mRNA、PIK3R1 mRNA、MAPK1 mRNA、MAPK3 mRNA水平。结果：参苓白术散组方中药活性成分204个，相关靶点335个；C-IN疾病相关靶点1 298个，交集靶点137个。利用Cytoscape 3.8.2软件分析参苓白术散“有效成分-交集靶点”网络得到排名前3位的核心活性成分为槲皮素、山柰酚、木犀草素。PPI网络分析筛选出核心靶点分别为TP53、STAT3、AKT1、HSP90AA1等。GO富集分析得到794项生物学过程、84项细胞组分及152项分子功能。KEGG富集分析共得到188条通路，主要涉及癌症相关通路、脂质和动脉粥样硬化相关通路以及糖尿病并发症的AGE-RAGE通路、PI3K/AKT信号通路和MAPK信号通路。核心活性成分与关键作用靶点分子对接显示结合性良好，且核心活性成分均与MAPK1蛋白有最优结合活性。与模型组比较，减毒组肾组织内铂离子含量明显降低（P<0.01）,BUN水平显著降低（P<0.05）,肾小管评分显著降低（P<0.01）,肾组织损伤明显改善，PIK3R1 mRNA表达水平显著升高（P<0.01）,TP53 mRNA、MAPK1 mRNA、MAPK3 mRNA表达水平显著降低（P<0.01）。结论：参苓白术散可能通过调节PI3K/AKT信号通路、MAPK信号通路缓解顺铂所致肾毒性。更多还原

【Abstract】 Objective: Based on network pharmacology, molecular docking technology, and animal experiments, to explore the core active ingredients and potential mechanism of Shenling Baizhu Powder in alleviating cisplatin-induced nephrotoxicity（C-IN）.Method: The active components of Shenling Baizhu Powder were preliminarily obtained and their targets were predicted through databases such as the Traditional Chinese Medicine System Pharmacology Database（TCMSP） and literature retrieval.Utilizing the Online Mendelian Inheritance in Man（OMIM）,the human gene database（GeneCards）,and the Drugbank database to obtain the targets of C-IN.The active component targets of Shenling Baizhu Powder and C-IN disease-related targets were input into Venny 2.1.0 software, and a Venn diagram was drawn to obtain the intersection targets of the two.The STRING database was used to build a PPI network of intersection targets and perform topology analysis to screen core targets.The DAVID data analysis platform was to conduct gene ontology（GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway enrichment analysis.Finally, select key targets and core active ingredients, and use AutoDock software for molecular docking.24 male C57BL/6J mice of SPF grade were randomly divided into control group, model group, attenuated group, and Shenling Baizhu Powder group, with 6 mice in each group.The mice in the model group and the attenuated group were intraperitoneally injected with cisplatin injection(3 mg·kg^-1) for every 3 days to induce renal injury, and the mice in the attenuated group were given Shenling Baizhu Powder(3 g·kg^-1) by intragastric administration at the same time.The Shenling Baizhu Powder was only given Shenling Baizhu Powder(3 g·kg^-1) by intragastric administration, and the control group was given the same amount of normal saline every day.The mice were sacrificed 30 days later, and the content of platinum ions in kidney tissue was detected by inductively coupled plasma mass spectrometry（ICP-MS）.The level of serum urea nitrogen（BUN） was measured by the urease method.The level of serum creatinine（Cre） was determined by the sarcosine oxidase method.The pathological changes of renal tissue were observed by HE staining.And the levels of AKT1mRNA,TP53 mRNA,PIK3R1 mRNA,MAPK1 mRNA,and MAPK3 mRNA in renal tissue were detected by PCR.Results: In Shenling Baizhu Powder prescription, there were 204 active ingredients, 335 related targets, 1 298 C-IN disease-related targets, and 137 intersecting targets.Using Cytoscape 3.8.2 software to analyze Shenling Baizhu Powder′s "active ingredient-intersection target" network, the top three core active ingredients were quercetin, kaempferol, and luteolin.The core targets selected by PPI network analysis were TP53,STAT3,AKT1,HSP90AA1,etc.GO enrichment analysis obtained 794 biological processes, 84 cellular components, and 152 molecular functions.KEGG enrichment analysis obtained a total of 188 pathways, mainly involving cancer-related pathways, lipid and atherosclerosis-related pathways and the AGE-RAGE signaling pathway in diabetic complications, PI3K/Akt signaling pathway, and MAPK signaling pathway.The docking of the core active ingredients and the key target molecules showed good binding, and the core active ingredients all had optimal binding activity with the MAPK1 protein.Compared with the model group, the platinum ion content in the kidney tissue of the attenuated group was significantly lower（P<0.01）,the BUN level was significantly lower（P<0.05）,the renal tubular score was significantly lower（P<0.01）,and the renal tissue damage was significantly alleviated.The expression level of PIK3R1 mRNA was significantly increased（P<0.01）,and the expression level of TP53 mRNA,MAPK1 mRNA,and MAPK3 mRNA was significantly decreased（P<0.01）.Conclusion: Shenling Baizhu Powder may alleviate C-IN by regulating the PI3K/Akt signaling pathway and MAPK signaling pathway.更多还原