益气养阴化浊通络方对高糖诱导小鼠肾足细胞自噬ATG5、Beclin-1及BNIP3表达的影响  

Effects of Yiqi Yangyin Huazhuo Tongluo Recipe on the expression of ATG5、Beclin-1 and BNIP3 in mouse renal podocytes autophagy induced by high glucose

作　　者：郭克磊 李颖利 韩立 张鹏[1,2] 叶松山 李凯[1,2] 卞华 GUO Kelei;LI Yingli;HAN Li;ZHANG Peng;YE Songshan;LI Kai;BIAN Hua(ZHANG Zhong-jing School of Chinese Medicine,Nanyang Institute of Technology,Henan Nanyang 473004,China;Henan Key Laboratory of ZHANG Zhong-jing Formulae and Herbs for Immunoregulation,Nanyang Institute of Technology,Henan Nanyang 473004,China)

机构地区：[1]南阳理工学院张仲景国医国药学院,河南南阳473004 [2]南阳理工学院河南省张仲景方药与免疫调节重点实验室,河南南阳473004

出　　处：《中国医院药学杂志》2023年第16期1808-1813,共6页Chinese Journal of Hospital Pharmacy

基　　金：河南省中医药科学研究专项课题(编号:20-21ZY1069);南阳市科技攻关项目(编号:KJGG104);南阳理工学院大学生科研基金项目(编号:NGDJ-2022-37)。

摘　　要：目的:观察益气养阴化浊通络方对高糖诱导的小鼠肾足细胞自噬相关蛋白5(autophagy-related protein 5,ATG5)、B细胞淋巴瘤-2蛋白相互作用中心卷曲螺旋蛋白1(B-cell lymphoma-2-interacting myosin-like coiled-coil protein 1,Beclin-1)及B淋巴细胞瘤-2基因/腺病毒E1B相互作用蛋白3(B-cell lymphoma-2/adenovirus E1 B interacting protein 3,BNIP3)表达的影响,探讨其对自噬的作用。方法:选取Wistar大鼠,分别灌胃20,40,80 g·kg^(-1)益气养阴化浊通络方及生理盐水,制备低、中、高浓度含药血清和空白血清。体外培养小鼠肾足细胞,分为正常对照组、高糖组、益气养阴化浊通络方低、中、高剂量组和雷帕霉素组。CCK-8法检测细胞增殖,细胞划痕检测细胞迁移能力,qRT-PCR和Western blot检测微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3 B,LC3B)、ATG5、Beclin-1及BNIP3 mRNA和蛋白表达。结果:与空白对照组相比,高糖组足细胞增殖减弱,迁移能力增强,LC3B、ATG5、Beclin-1及BNIP3 mRNA和蛋白表达显著减弱;与HG组相比,益气养阴化浊通络方低、中、高剂量含药血清组及雷帕霉素组均可促进足细胞增殖,抑制足细胞迁移,明显促进LC3B、ATG5、Beclin-1及BNIP3 mRNA和蛋白表达。结论:益气养阴化浊通络方能够促进高糖诱导的小鼠肾足细胞自噬,调节足细胞增殖及迁移。OBJECTIVE To observe the effects of Yiqi Yangyin Huazhuo Tongluo Recipe on the expression of autophagy-related protein 5(ATG5)、Beclin-1 and B-cell lymphoma-2/adenovirus E1 B interacting protein 3(BNIP3)in mouse renal podocytes induced by high glucose,and to explore its effect on autophagy.METHODS Wistar rats were given 20,40 and 80 g·kg^(-1) of Yiqi Yangyin Huazhuo Tongluo Recipe and normal saline to prepare low,medium and high concentration drug-containing serum and blank serum.Mouse renal podocytes were cultured in vitro and divided into normal control group,high glucose group,Yiqi Yangyin Huazhuo Tongluo Recipeon low,medium and high dose groups,and rapamycin group.Cell proliferation was detected by CCK-8,cell migration ability was detected by cell scratching,mRNA and protein expressions of LC3,ATG5,Beclin-1 and BNIP3 were detected by qRT-PCR and Western blot.RESULTS Compared with the normal control group,the proliferation of podocytes in the high glucose group was weakened,the migration ability was enhanced,and the mRNA and protein expressions of LC3,ATG5,Beclin-1 and BNIP3 were significantly reduced;Yiqi Yangyin Huazhuo Tongluo Recipeon low,medium and high dose groups and rapamycin group could promote the proliferation of podocytes,inhibit the migration of podocytes,and significantly promote the mRNA and protein expressions of LC3,ATG5,Beclin-1 and BNIP3.CONCLUSION Yiqi Yangyin Huazhuo Tongluo Recipe can promote high glucose-induced autophagy in mouse renal podocytes and regulate podocyte proliferation and migration.

关 键 词：糖尿病肾病 益气养阴化浊通络方 自噬 小鼠肾足细胞 自噬相关蛋白5 B淋巴细胞瘤-2基因/腺病毒E1B相互作用蛋白3 B细胞淋巴瘤-2蛋白相互作用中心卷曲螺旋蛋白1 

分 类 号：R285.5[医药卫生—中药学]