文献类型：期刊文献

中文题名：武威汉简瘀方靶向Sirt1抑制PI3K/AKT信号通路对高血压肾损伤的影响

英文题名：Effect of Wuwei Hanjian Yu Fang Targeting Sirt1 to Inhibit PI3K/AKT Signaling Pathway on Hypertensive Renal Injury

作者：沈路凡[1,2];徐静[1,2];马睿玲[1,2];侯明双[1,2];吕红英[1,2];贾冠军[1,2];寇雨顺[1,2];康万荣[1];伊琳[1,2]

第一作者：沈路凡

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医药防治慢性病疾病重点实验室,兰州730000

第一机构：甘肃中医药大学

年份：2025

卷号：41

期号：11

起止页码：21

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金资助项目(编号:82160842);甘肃省中医药科研项目(编号:GZKP-2022-40);甘肃省联合科研基金一般项目(编号:23JRRA1521)。

语种：中文

中文关键词：武威汉简瘀方;高血压肾损伤;网络药理学;蛋白激酶;磷脂酰肌醇3-激酶;鼠双微体基因2蛋白;沉默调节蛋白1

外文关键词：Wuwei Hanjian Yu Fang(武威汉简瘀方);Hypertensive renal injury;Network pharmacology;Protein kinase;Phosphatidylinositol 3-kinase;Murine double minute 2;Silent information regulator 1

摘要：目的:通过网络药理学和分子对接技术预测并筛选武威汉简瘀方干预高血压肾损伤的潜在分子靶点,通过动物实验分析瘀方对高血压肾损伤潜在靶点及通路表达的影响。方法:借助中药系统药理学数据库TCMSP以及PubChem获取武威汉简瘀方的有效活性成分并进行靶标转换,利用Gene Cards、OMIM数据库获取高血压肾损伤靶点;取武威汉简瘀方与高血压肾损伤交集靶点,构建交集靶点互作网络,并进行拓扑分析,得到关键靶点;运用Auto Dock软件将关键化合物与高血压肾损伤关键蛋白受体进行分子对接;对关键靶点进行GO功能分析和KEGG通路富集分析;以武威汉简瘀方颗粒剂灌胃自发性高血压大鼠(SHR),连续8 w后,检测大鼠血压、尿蛋白、肌酐、尿素氮含量;透射电镜观察大鼠肾脏组织形态改变;Western blot、RT-PCR法检测筛选得到的关键靶点以及相关信号通路蛋白的表达。结果:武威汉简瘀方的主要活性成分为槲皮素、山柰酚、β-谷甾醇等,与Mdm2、Akt1、Sirt1等关键靶点对接良好,其中以槲皮素与各个靶点间的结合情况最为稳定。关键靶点主要参与调节自噬、巨噬细胞的负调控等生物过程,并富集于CellμLar senescence、PI3K、AKT、AMPK和P53等信号通路。动物实验研究结果显示,与正常对照组相比,模型对照组大鼠的血压显著升高(P<0.01),尿蛋白、尿肌酐、尿素氮含量显著升高(P<0.01),肾小球和肾小管内血管内皮的损伤加重,肾组织Mdm2、Akt1的mRNA表达上调,Sirt1mRNA表达下调(P<0.01),p-PI3K、p-AKT1和MDM2蛋白表达上调,SIRT1蛋白表达下调(P<0.01);与模型对照组相比,瘀方干预后大鼠血压明显下降(P<0.05或P<0.01),尿蛋白、尿肌酐、尿素氮含量降低(P<0.05或P<0.01),肾组织Mdm2与Akt1的mRNA表达下调,Sirt1mRNA表达上调(P<0.05或P<0.01),p-PI3K、p-AKT1和MDM2蛋白表达下调,SIRT1蛋白表达上调(P<0.05或P<0.01)。结论:武威汉简瘀方对高血压肾损伤具有一定的保护作用,可能通过上调Sirt1的表达抑制PI3K/AKT/MDM2信号通路发挥保护作用。
Objective:To predict and screen the potential molecular targets of Wuwei Hanjian Yu Fang(武威汉简瘀方)for intervening in hyper-tensive renal injury through network pharmacology and molecular docking technology and analyze the effects of the formula on the expression of potential targets and pathways in hypertensive renal injury through animal experiments.Methods:The TCMSP and PubChem databases were used to obtain active ingredients of Wuwei Hanjian Yu Fang,and target conversion was carried out.The Gene Cards and OMIM databases were used to obtain the targets of hypertensive renal injury.The intersection targets of Wuwei Hanjian Yu Fang and hypertensive renal injury were employed to construct an intersection target interaction network and topology analysis and get key targets.The Auto Dock software was used for the molecular docking of key compounds and key protein receptors of hypertensive renal injury,and the GO function and KEGG path-way enrichment were analyzed on key targets.The results of network pharmacology and molecular docking above were verified via the animal experiment.Wuwei Hanjian Yu Fang granules were orally administrated on spontaneously hypertensive rats(SHR)for eight weeks,and then the levels of blood pressure,urinary protein,creatinine,and urea nitrogen were detected.The morphological changes of renal tissue in SHR rats were observed by transmission electron microscopy,and the expression levels of the screened key targets and related signaling pathways were detected by Western Blot and RT-PCR.Results:Quercetin,kaempferol,β-sitosterol,and other main active ingredients of Wuwei Hanjian Yu Fang were docked well with Mdm2,Akt1,Sirt1,and other key targets.Specifically,quercetin showed the most stable binding with these tar-gets.The key targets were mainly involved in the regulation of autophagy,negative regulation of macrophage,and other biological processes and were enriched in the signaling pathways such as CellμLar senescence,PI3K-AKT,AMPK,and P53.Animal experimental results showed that compared with the normal control group,the model control group displayed significantly elevated blood pressure level(P<0.01),signifi-cantly increased contents of urinary protein,creatinine,and urea nitrogen(P<0.01),exacerbated endothelial damage in the glomerular and tubular vasculature,up-regulated Mdm2 and Akt1 mRNA expressions in renal tissue,down-regulated Sirt1 mRNA expression(P<0.01),in-creased P-PI3K,P-AKT1,and MDM2 protein expressions,and decreased SIRT1 protein expression(P<0.01).Compared with the model control group,the rats after the intervention of the formula displayed significantly decreased blood pressure levels(P<0.01 or P<0.05),re-duced contents of urinary protein,creatinine,and urea nitrogen(P<0.01 or P<0.05),down-regulated Mdm2 and Akt1 mRNA expressions in renal tissue,up-regulated Sirt1 mRNA expression(P<0.01 or P<0.05),decreased P-PI3K,P-AKT1,and MDM2 protein expressions,and in-creased SIRT1 protein expression(P<0.01 or P<0.05).Conclusion:Wuwei Hanjian Yu Fang has a certain protective effect on hypertensive renal injury,which may be exerted through up-regulation of Sirt1 expression by quercetin as the core active ingredient to inhibit the PI3K-AKT-MDM2 signaling pathway.