芪地糖肾方调控AMPK/ULK1信号通路激活自噬对高糖诱导足细胞凋亡的影响  

Effects of Qi-Di-Tang-Shen decoction inhibits high glucose-induced apoptosis of podocytes via activating autophagy through regulating AMPK/ULK1 signaling pathway

作　　者：郭燕 柳红芳[1] 史扬[2] 郑毅成 宿家铭 董昭熙 刘家佑 仝伶秀 GUO Yan;LIU Hongfang;SHI Yang;ZHENG Yicheng;SU Jiaming;DONG Zhaoxi;LIU Jiayou;TONG Lingxiu(Dongzhimen Hospital of Beijing University of Chinese Medicine,Beijing 100700,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China;Beijing Chaoyang Emergency Rescue Hospital of Integrated Traditional Chinese and Western Medicine,Beijing 100000,China)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]河南中医药大学第一附属医院,河南郑州450000 [3]北京朝阳中西医结合急诊抢救医院,北京100000

出　　处：《现代中西医结合杂志》2025年第1期18-23,31,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：北京市自然科学基金资助项目(7212180);北京中医药大学东直门医院科技创新项目(DZMKJCX-2022-003,DZMKJCX-2023-005)。

摘　　要：目的基于腺苷酸活化蛋白激酶/unc-51样激酶1(AMPK/ULK1)信号通路激活自噬探讨芪地糖肾方对高糖诱导足细胞凋亡的影响。方法取小鼠肾小球足细胞,分为正常糖组、高糖组、高糖+芪地糖肾方组,分别予以5.5 mmol/L完全培养基、35 mmol/L高糖培养基、35 mmol/L高糖培养基+芪地糖肾方冻干粉200μg/mL培养48 h。采用免疫荧光法观察细胞形态及细胞中p-AMPK、ULK1蛋白表达情况,采用Western blot法检测细胞中AMPK、p-AMPK、ULK1、微管相关蛋白1(Beclin1)、p62、微管相关蛋白1轻链3(LC3)、白血病基因-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、肾素(Nephrin)蛋白表达情况。结果与正常糖组比较,高糖组细胞骨架损伤,细胞中p-AMPK、ULK1表达荧光强度均明显降低(P均<0.05);细胞中p-AMPK/AMPK、ULK1、Beclin1、LC3、Bcl-2、Nephrin蛋白相对表达量均明显降低(P均<0.05),p62、Bax蛋白相对表达量均明显升高(P均<0.05)。与高糖组比较,高糖+芪地糖肾方组细胞骨架损伤减轻,细胞中p-AMPK、ULK1表达荧光强度均明显升高(P均<0.05);细胞中p-AMPK/AMPK、ULK1、Beclin1、LC3、Bcl-2、Nephrin蛋白相对表达量均明显升高(P均<0.05),p62、Bax蛋白相对表达量均明显降低(P均<0.05)。结论芪地糖肾方可以通过调控AMPK/ULK1信号通路激活自噬而抑制高糖诱导的足细胞凋亡。Objective It is to investigate the effect of Qi-Di-Tang-Shen decoction(QDTSD)on high glucose-induced apoptosis of podocytes via activating autophagy through regulating AMP activated protein kinase and unc-51-like kinase 1(AMPK/ULK1)signaling pathway.Methods Mouse glomerular podocytes were taken and divided into normal glucose group,high glucose group and high glucose+QDTSD group,and were cultured in 5.5 mmol/L complete medium,35 mmol/L high glucose medium,35 mmol/L high glucose medium+200μg/mL lyophilized powder of QDTSD for 48 h,respectively.The cell morphology and the protein expressions of pAMPK and ULK1 in the cells were observed by immunofluorescence method,the protein expressions of AMPK,pAMPK,ULK1,Beclin1,p62,LC3,Bcl-2,Bax and nephrin were detected by Western blot method.Results Compared with the normal glucose group,the cytoskeleton was damaged in the high glucose group,and the fluorescence intensities of p-AMPK and ULK1 expressions in the cells were all significantly reduced(all P<0.05);the relative protein expressions of p-AMPK/AMPK,ULK1,Beclin1,LC3,Bcl-2,and Nephrin in the cells were all significantly reduced(P all<0.05),while the protein relative expressions of p62 and Bax were all significantly increased(all P<0.05).Compared with the high glucose group,the cytoskeleton damage was reduced in the high glucose+QDTSD group,and the fluorescence intensities of the expressions of p-AMPK and ULK1 in the cells were significantly increased(all P<0.05);the relative protein expressions of p-AMPK/AMPK,ULK1,Beclin1,LC3,Bcl-2,Nephrin in the cells was significantly increased(all P<0.05),and the relative protein expressions of p62 and Bax were all significantly decreased(all P<0.05).Conclusion QDTSD can inhibit the apoptosis of podocytes induced by high glucose via activating autophagy through regulating AMPK/ULK1 signaling pathway.

关 键 词：糖尿病肾脏病 足细胞 芪地糖肾方 腺苷酸活化蛋白激酶 unc-51样激酶1 自噬 凋亡 

分 类 号：R-33[医药卫生]