摘要
目的探究小檗碱对高糖诱导下足细胞铁死亡的作用机制。方法Western blot法和RT-qPCR法检测desmin、podocin、GPX4、PTGS2、ACSL4在高糖刺激0 h、6 h、12 h、24 h、36 h的变化情况以及小檗碱对高糖情况下Nrf2、HO-1、GPX4和podocin的影响;EdU观察高糖刺激不同时间足细胞的增殖情况;CCK-8法测定足细胞的活力;荧光倒置显微镜,GSH和GSSG试剂盒检测小檗碱对高糖诱导下足细胞氧化应激水平的影响;激光共聚焦显微镜观察desmin的表达;透射电子显微镜观察足细胞的超微形态特征。结果Podocin和GPX4蛋白表达量在24 h明显降低,ACSL4和PTGS2的mRNA水平显著上调。小檗碱明显改善ROS和GSH的水平,上调Nrf2、HO-1、GPX4和podocin的表达,降低PTGS2和ACSL4的水平,从而缓解高糖情况下足细胞质膜起泡、线粒体皱缩等变化。结论在高糖环境下,足细胞会发生不同于自噬、凋亡的另一种数量减少的现象,即铁死亡。小檗碱可以缓解此现象的发生,其机制可能与Nrf2/HO-1/GPX4通路有关。
Aim To explore the regulatory mechanism of berberine in the high glucose(HG)-induced podocyte ferroptosis.Methods Western blot and RT-qPCR were used to detect the changes of GPX4,PTGS2,ACSL4,podocin,and desmin at different time(0 h,6 h,12 h,24 h,36 h)of HG stimulation,and the relative expression of Nrf2,HO-1,GPX4 and podocin under HG after treated with berberine.The proliferation of podocytes stimulated by HG at different time points was observed by EdU staining.The therapeutic effect of berberine on podocyte damage was screened by CCK-8,and the effect of berberine on the level of oxidative stress in HG-induced podocytes were measured by fluorescence inverted microscope,GSH and GSSG kits.In addition,transmission electron microscopy was employed to observe the ultrastructural characteristics of podocytes in each group.Results The expression of podocin and GPX4 was significantly reduced at 24 h,and the mRNA levels of ACSL4 and PTGS2 were significantly up-regulated.Berberine could notably increase the expression of Nrf2,HO-1,GPX4 and podocin,and reduce the levels of PTGS2 and ACSL4.Moreover,berberine significantly improved the levels of ROS and GSH in podocytes under HG conditions,thereby alleviating membrane blistering,mitochondrial shrinkage and other morphological changes of HG-induced podocytes.Conclusions In this study,the number of podocytes decreases,which is a death mode different from autophagy and apoptosis,that is,ferroptosis.Berberine can alleviate the occurrence of this phenomenon by mediating the Nrf2/HO-1/GPX4.
作者
关锡梅
解勇圣
倪伟建
唐丽琴
GUAN Xi-mei;XIE Yong-sheng;NI Wei-jian;TANG Li-qin(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230001,China;School of Pharmacy,Anhui Medical University,Hefei 230032,China;the First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230001,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第3期396-403,共8页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81803602,81773955)
安徽省自然科学基金资助项目(No 1708085QH207)
中央高校基本科研业务费专项资金资助项目(No WK9110000018)。
关键词
小檗碱
糖尿病肾病
足细胞
氧化应激
铁死亡
berberine
diabetic nephropathy
podocytes
oxidative stress
ferroptosis
作者简介
关锡梅(1995-),女,硕士生,研究方向:内分泌及代谢药理学,E-mail:18297911500@163.com;通讯作者:倪伟建(1990-),男,主管药师,硕士生导师,研究方向:内分泌及代谢药理学,E-mail:niweijian@ustc.edu.cn;通讯作者:唐丽琴(1972-),女,主任药师,教授,博士生导师,研究方向:内分泌及代谢药理学,E-mail:tangliqin@ustc.edu.cn。
