小檗碱对高糖诱导足细胞凋亡的干预作用及其相关调控机制研究  

Effects of berberine on high glucose-induced podocyte apoptosis and its related mechanism

作　　者：李超 关锡梅 倪伟建 唐丽琴 LI Chao;GUAN Xi-mei;NI Wei-jian;TANG Li-qin(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Dept of Pharmacy,Anhui Provincial Hospital,the First Affiliated Hospital of University of Science and Technology of China,Hefei 230001,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]中国科学技术大学附属第一医院(安徽省立医院)药剂科,安徽合肥230001

出　　处：《中国药理学通报》2019年第12期1681-1687,共7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)

摘　　要：目的研究高糖对小鼠肾小球足细胞凋亡的影响及其特点,探讨小檗碱对高糖诱导足细胞凋亡的干预作用及可能机制。方法体外培养小鼠肾小球足细胞,观察足细胞在不同浓度高糖和不同培养时间的形态学变化,CCK-8法测定足细胞的活性,Hoechst染色、Annexin V-FITC/PI双染法检测足细胞的凋亡水平,激光共聚焦显微镜观察caspase-3的表达,Western blot法检测caspase-3、6、7、8、9与Bcl-2的表达。结果高糖(15、20、25、30 mmol·L^-1)培养分别在72、48、36、24 h时诱发足细胞凋亡;小檗碱能增加高糖培养的足细胞活力,明显减少高糖诱导的足细胞凋亡,不同程度的下调高糖诱导的caspase-3、6、7、8和9的表达,提高Bcl-2表达。结论研究不同高糖环境在不同时间诱发足细胞凋亡的特点,为进行足细胞凋亡与肾脏损伤的干预治疗提供重要的时间参考;小檗碱缓解高糖诱发的足细胞凋亡,其作用机制可能与caspase-8/caspase-3和Bcl-2/caspase-9/caspase-3凋亡通路相关。Aim To investigate the influence and characteristics of high glucose(HG)induced podocyte ap-optosis,and explore the protective effect of berberine on the apoptosis and the possible mechanism.Methods Mouse podocytes were cultured under different time and HG conditions to observe the effects of different glucose levels and time on podocyte apoptosis.The effect of berberine on podocyte activity was measured by CCK-8 assay.Meantime,the protective effect of berberine on podocyte apoptosis was detected by Hoechst staining and flow cytometry.In addition,the effects of HG and berberine on the expression of apoptotic proteins and related signaling pathways in podocytes were analysed by laser confocal microscopy and Western blot.Results High glucose(15,20,25 and 30 mmol·L^-1)could induce podocyte apoptosis at 72,48,36 and 24h,respectively.Berberine could increase the activity of podocytes cultured with high glucose,significantly reduce the apoptosis of podocytes induced by high glucose,down-regulate the expression of caspase-3,6,7,8 and 9 induced by high glucose,and increase the expression of Bcl-2.Conclusions The characteristics of podocyte apoptosis induced by different HG and time may provide important reference for clinical intervention of podocyte apoptosis and kidney injury.Berberine can alleviate the apoptosis of podocyte induced by high glucose,and its mechanism may be related to the apoptosis pathway of caspase-8/caspase-3 and Bcl-2/caspase-9/caspase-3 signaling pathways.

关 键 词：小檗碱 糖尿病肾病 高糖 足细胞凋亡 凋亡信号通路 干预作用 

分 类 号：R-332[医药卫生] R282.71