缩泉益肾方含药血清通过抑制KDM6A调节肾小球足细胞上皮-间充质转化
Regulation of Epithelial-Mesenchymal Transition of Glomerular Podocytes by Suoquan Yishen(缩泉益肾)Formula Containing Serum through Inhibition of KDM6A
王菲 1肖曼 1陈柏岑 1黄明月 1马妍 1刘书蔓 1王珏 2夏玉成 1谢毅强 1赵明明1
作者信息
- 1. 海南医学院,海口 571199
- 2. 贵州中医药大学,贵阳 550025
- 折叠
摘要
目的:探讨缩泉益肾方含药血清通过抑制组蛋白去甲基化酶(Histone demethylase,KDM6A),调控上皮-间充质转化(Epithelial-mesenchymal transition,EMT)相关标志物和足细胞损伤因子的表达,改善足细胞(Podocyte)EMT和足细胞损伤的机制研究.方法:除正常对照组外,高糖胁迫足细胞损伤后,随机分为模型对照组、缩泉益肾方14.4 g/kg含药血清5％、10％、20％组和卡格列净4.45 μg/mL组.采用显微镜观察足细胞形态实验、CCK-8法、细胞划痕实验和流式细胞术,分别检测足细胞形态变化、活力、迁移力和凋亡率;RT-qPCR和Western blot检测足细胞中组蛋白去甲基化酶(Kdm6a)、Krüppel样因子10(Klf10)、足细胞裂孔膜蛋白(Nephrin)、α-平滑肌肌动蛋白(Asma)、N-钙粘蛋白(Ncadherin)和P-钙粘蛋白(Pcadherin)mRNA和蛋白的表达.结果:与正常对照组比较,模型对照组足细胞活力、迁移力显著降低,凋亡率显著升高(P＜0.01),Kdm6a、Klf10、Asma、Ncadherin mRNA和蛋白的表达显著上调(P＜0.01),Pcadherin、Nephrin mRNA和蛋白的表达显著下调(P＜0.01);与模型对照组比较,缩泉益肾方14.4 g/kg含药血清5％、10％和卡格列净4.45 μg/mL组足细胞活力、细胞迁移力显著升高(P＜0.01),细胞凋亡率显著降低(P＜0.01),缩泉益肾方14.4 g/kg含药血清10％组和卡格列净4.45 μg/mL组中Kdm6a、Klf10、Asma、Ncadherin mRNA和蛋白的表达下调(P＜0.05或P＞0.01),Pcadherin、Nephrin mRNA和蛋白的表达明显上调(P＜0.05或P＜0.01).结论:缩泉益肾方含药血清可能通过抑制KDM6A,调控EMT相关标志物和足细胞损伤因子,从而改善足细胞EMT和足细胞损伤.
Abstract
Objective:To investigate the mechanism by which Suoquan Yishen(缩泉益肾)Formula containing serum improves epithelial-mesen-chymal transition(EMT)and injury of podocytes by inhibiting the expression of histone demethylase KDM6A and regulating EMT-related markers and podocyte injury factors.Methods:Except for the normal control group,after podocyte injury by high glucose stress,the experiment was randomly divided into model control group,14.4 g/kg Suoquan Yishen Formula containing serum of 5％,10％,and 20％groups,and 4.45 μg/mL Cagliflozin group.The morphological changes,viability,mobility,and apoptosis rate of the podocyte were detected by podocyte morphology assay,CCK-8 method,cell scratch assay,and flow cytometry,respectively.RT-qPCR and Western blot were used to detect the mR-NA and protein expressions of Kdm6a,Krüppel-like factor 10(Klf10),podocyte lytic membrane protein(Nephrin),α-smooth muscle actin(α-Sma),N-cadherin(N-cadherin),and P-cadherin(P-cadherin).Results:Compared with the normal control group,the model control group significantly decreased podocyte viability and mobility and increased apoptosis rate(P＜0.01).The mRNA and protein expressions of Kdm6a,Klf10,α-Sma and N-cadherin were significantly up-regulated(P＜0.01),while the mRNA and protein expressions of P-cadherin and Nephrin were significantly down-regulated(P＜0.01).Compared with the model control group,podocyte viability and mobility were significant-ly increased(P＜0.01),and apoptosis rate was significantly decreased(P＜0.01)in the 14.4 g/kg Suoquan Yishen Formula containing ser-um of 5％and 10％groups and 4.45 μg/mL Cagliflozin group.In the 14.4 g/kg Suoquan Yishen Formula containing serum of 10％group and 4.45 μg/mL Cagliflozin group,the mRNA and protein expressions of Kdm6a,Klf10,α-Sma and N-cadherin were down-regulated(P＜0.01 or P＞0.05),while those of P-cadherin and Nephrin were significantly up-regulated(P＜0.05 or P＜0.01).Conclusion:Suoquan Yish-en Formula containing serum may improve EMT and injury of podocytes by inhibiting KDM6A and regulating EMT-related markers and podo-cyte injury factors.
关键词
缩泉益肾方/糖尿病肾病/足细胞/上皮-间充质转化/组蛋白去甲基化酶Key words
Suoquan Yishen(缩泉益肾)Formula/Diabetic nephropathy/Podocyte/Epithelial-mesenchymal transition/Histone demethylase KDM6A引用本文复制引用
基金项目
国家自然科学基金面上项目(82174334)
海南省研究生创新科研项目(Qhys2021-358)
出版年
2024
国家科技期刊平台
NSTL
万方数据