中药药理与临床2025，Vol.41Issue(10)：2-10,9.
补阳还五汤调控JAK2/STAT3信号通路改善肾小管上皮细胞线粒体稳态的机制研究
Improvement of Buyang Huanwu(补阳还五)Decoction on Mitochondrial Homeostasis of Renal Tubule Epithelial Cells Based on JAK2/STAT3 Signaling Pathway
摘要
Abstract
Objective:To investigate the mechanism of Buyang Huanwu(补阳还五)Decoction in regulating the mitochondrial homeostasis of hu-man proximal renal tubular epithelial cells(HK-2)induced by high glucose through the JAK2/STAT3 signaling pathway.Methods:Thirty SD rats were randomly divided into a control group and a 14.98 g/kg Buyang Huanwu Decoction group,with 15 rats in each group.Rats were administered with corresponding drugs or saline solution via gavage for seven consecutive days.Drug-containing serum and normal control ser-um were collected afterwards.HK-2 cells were further divided into normal control serum group,model control group,Empagliflozin group,Buy-ang Huanwu Decoction group,hyperosmolar control group,and JAK pathway inhibitor AG490 group.CCK8 assay was used to screen for a 10%concentration of drug-containing serum and normal control serum for 48 hours of intervention.Western blot,real-time quantitative PCR,and immunofluorescence were performed to detect the expressions of KIM-1,inflammatory factors,apoptosis-related proteins,pathway-related proteins,and mitochondrial dynamics proteins.MDA,SOD,ROS,and ATP levels were measured using ELISA.Mitochondrial morphology was observed by transmission electron microscopy,and mitochondrial membrane potential was assessed by flow cytometry.Results:Compared with those in the normal control serum group,the expressions of KIM-1,IL-6,IL-1β,and TNF-α in the model control group were significantly in-creased(P<0.01),accompanied by increased expressions of apoptotic proteins Bax and C-caspase3(P<0.01)and down-regulated expres-sion of anti-apoptotic protein BCL-2.The contents of oxidative stress marker MDA and ROS were significantly elevated(P<0.01),while ATP content and SOD activity were significantly decreased(P<0.01).Protein expressions of p-JAK2 and p-STAT3 were up-regulated(P<0.01),and the expressions of DRP1 and FIS1 were up-regulated(P<0.01),while the expressions of fusion proteins OPA1 and MFN2 were sig-nificantly down-regulated(P<0.01).The mitochondrial membrane potential was decreased(P<0.01),and transmission electron microscopy showed disrupted mitochondrial morphology,including cristae disruption,vacuolar degeneration,swelling,and fragmentation.Compared with those of the model control group,the contents of HK-2 cells KIM-1 and inflammatory factors were decreased in 10%drug-containing serum of the Buyang Huanwu Decoction group,and the expression of the apoptotic protein was down-regulated.The contents of ROS and MDA were de-creased,and the ATP and SOD synthesis were increased,the protein expressions of p-JAK2 and p-STAT3 were down-regulated(P<0.01),and the expressions of DRP1 and FIS1 were down-regulated(P<0.01).The mitochondrial membrane potential was increased,while the mito-chondrial morphology was partially restored.Conclusion:Buyang Huanwu Decoction can alleviate inflammation and oxidative stress of human proximal renal tubular epithelial cells cultured under high glucose conditions.It can also inhibit cell apoptosis,improve damaged mitochondri-al morphology and membrane potential,and maintain mitochondrial dynamics stability.These effects may be related to its negative regulation of the JAK2/STAT3 signaling pathway.关键词
补阳还五汤/高糖/人近端肾小管上皮细胞/线粒体/Janus激酶信号转导器2/转录的信号转导和激活因子3通路Key words
Buyang Huanwu(补阳还五)Decoction/High glucose/Human proximal renal tubular epithelial cell/Mitochondria/JAK2/STAT3 pathway引用本文复制引用
娄菲菲,杨帆,陈志强..补阳还五汤调控JAK2/STAT3信号通路改善肾小管上皮细胞线粒体稳态的机制研究[J].中药药理与临床,2025,41(10):2-10,9.基金项目
国家自然科学基金(编号:82274504) （编号:82274504）
河北省中医药管理局资助项目(编号:2021167). （编号:2021167）
