基于网络药理学探讨EGCG对顺铂所致大鼠急性肾损伤的保护作用

Protective effect of epigallocatechin-3-gallate on cisplatin-induced acute kidney injury in rats based on network pharmacology

目的基于网络药理学和体内动物模型实验,评价表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对顺铂(cisplatin,CIS)诱导的大鼠急性肾损伤(acute kidney injury,AKI)的保护效果。方法通过TCMSP、Gene Cards、OMIM网站收集EGCG、AKI作用靶点,取交集后构建蛋白质-蛋白质互作网络(PPI),Cytoscape 3.9.1对交集靶点进行可视化分析,筛选出关键靶点,通过DAVID数据库进行KEGG和GO富集分析。32只雄性Wistar大鼠,随机分为对照组(CON组)、EGCG组、顺铂组(CIS组)和CIS+EGCG组。CON组和CIS组灌胃生理盐水,EGCG组和CIS+EGCG组灌胃EGCG(40 mg/kg),连续28 d,第26天CIS组、CIS+EGCG组腹腔注射CIS(7 mg/kg),第29天收集血液和组织。检测血清尿素氮(BUN)、肌酐(SCr)水平;苏木素-伊红(HE)染色观察肾病理变化;TUNEL检测肾组织细胞凋亡情况;Western Blot、qRT-PCR及免疫组化验证分析结果。结果网络药理学筛选出87个EGCG与AKI交集基因,25个核心靶点,通过PI3K/AKT等信号通路和多种生物过程影响AKI的发展;EGCG预处理明显降低AKI大鼠血清中BUN、SCr水平,改善AKI大鼠肾病变,缓解AKI大鼠肾组织凋亡;Western Blot、qRT-PCR及免疫组化结果表明预处理EGCG可激活PI3K/AKT通路。结论基于以上研究结果,推测出EGCG可能通过激活PI3K/AKT信号通路进而缓解CIS诱导的大鼠AKI。

Objective To evaluate the protective effect of epigallocatechin-3-gallate(EGCG)on cisplatin(CIS)-induced acute kidney injury(AKI)in rats based on network pharmacology and in vivo animal model experiments.Methods Targets of EGCG and AKI were collected from the TCMSP,Gene Cards,and OMIM websites,and a protein-protein interaction network was constructed based on the intersection.The intersection targets were analyzed visually using Cytoscape 3.9.1 and the key targets were screened out.Kyoto Encyclopedia and of Genes and Genomes and Gene Ontology enrichment analyses were carried out using the DAVID database.Thirty-two male Wistar rats were divided randomly into four groups:CON group,EGCG group,CIS group,and CIS+EGCG group.Rats in the control and CIS groups were given normal saline every day,rats in the EGCG and CIS+EGCG groups were given EGCG(40 mg/kg)every day for 28 d,and rats in the CIS and CIS+EGCG groups received an intraperitoneal injection of CIS(7 mg/kg)on the 26th day.Blood and tissue samples were obtained on the 29th day.Serum urea nitrogen and creatinine levels were detected,renal pathology was observed by HE staining,and apoptosis in renal tissue was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling.Western Blot,qRT-PCR,and immunohistochemistry were used to verify the result.Results Eighty-seven genes intersecting EGCG and AKI and 25 core targets were screened from network pharmacology,which influenced the development of AKI via signal pathways such as PI3K/Akt and various biological processes.EGCG pretreatment significantly reduced serum levels of serum urea nitrogen and creatinine,improved the renal pathology,and reduced renal tissue apoptosis in AKI rats.Western Blot,qRT-PCR and immunohistochemistry showed that pretreatment with EGCG activated the PI3K/Akt pathway.Conclusions EGCG alleviates CIS-induced AKI in rats via the PI3K/Akt signaling pathway.