基于Nrf2/HO-1/GPX4信号轴探讨参芪地黄汤抑制高糖诱导人肾小管上皮细胞铁死亡的作用机制  

Shenqi Dihuang Decoction inhibits high-glucose induced ferroptosis of renal tubular epithelial cells via Nrf2/HO-1/GPX4 pathway

作　　者：王智槟 邹晓玲[1] 邹译娴[1] 王理槐[1] 吴源陶 WANG Zhi-bin;ZOU Xiao-ling;ZOU Yi-xian;WANG Li-huai;WU Yuan-tao(Department of Endocrinology,the First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Graduate School,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院内分泌科,湖南长沙410007 [2]湖南中医药大学研究生院,湖南长沙410208

出　　处：《中国中药杂志》2023年第19期5337-5344,共8页China Journal of Chinese Materia Medica

基　　金：湖南省中医药管理局项目(D2022101)。

摘　　要：为探讨参芪地黄汤对高糖诱导人肾小管上皮细胞(HK-2)铁死亡及核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase 1,HO-1)/谷胱甘肽过氧化物酶(glutathione peroxidase 4,GPX4)轴的影响及其潜在机制。构建高糖诱导HK-2细胞损伤模型,制备参芪地黄汤含药血清;筛选参芪地黄汤含药血清的最佳浓度及干预时间,将HK-2分为正常组,高糖组,参芪地黄汤低、中、高剂量组。各组细胞干预后,检测细胞增殖率,观察细胞形态和线粒体超微结构,检测细胞内活性氧(reactive oxygen species,ROS)、亚铁离子(Fe^(2+))、谷胱甘肽(glutathione,GSH)和丙二醛(malondialdehyde,MDA)水平,检测细胞内Nrf2、HO-1、GPX4和xCT蛋白的表达。筛选出10%参芪地黄汤干预24 h为最佳剂量与干预时间,与高糖组比较,参芪地黄汤高剂量组促进高糖诱导的HK-2增殖;与高糖组比较,参芪地黄汤低、中、高剂量组细胞排列紧密,细胞数量增加,改善梭形纤维样为鹅卵石样形态,且改善线粒体膜和嵴的形态结构;抑制ROS升高,改善过氧化损伤;降低Fe^(2+)、MDA水平,升高GSH水平,抑制铁死亡脂质过氧化损伤;激活抗氧化通路,上调Nrf2、HO-1、xCT和GPX4蛋白的表达。总之,参芪地黄汤含药血清在体外能抑制高糖诱导HK-2铁死亡,其机制与抗氧化作用和激活Nrf2/HO-1/GPX4信号轴有关。This study aims to explore the effects of Shenqi Dihuang Decoction on high-glucose induced ferroptosis and the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4) axis in human renal tubular epithelial cells(HK-2) and to clarify the underlying mechanism.The cell injury model was established by exposing HK-2 to high glucose,and the Shenqi Dihuang Decoction-medicated serum was prepared.The optimal concentration and intervention time of Shenqi Dihuang Decoction were determined.HK-2 were divided into normal,high glucose,and low-,medium-,and high-dose Shenqi Dihuang Decoction groups.After interventions,the cell proliferation rate in each group was determined and the cell morphology and mitochondrial ultrastructure were observed.Then,the levels of intracellular reactive oxygen species(ROS),ferrous ion(Fe^(2+)),glutathione(GSH),and malondialdehyde(MDA) and the protein levels of Nrf2,HO-1,GPX4,and xCT were measured.The optimal concentration and intervention time of Shenqi Dihuang Decoction-medicated serum were determined to be 10% and 24 h,respectively.Compared with the high glucose group,high-dose Shenqi Dihuang Decoction promoted the proliferation of HK-2.The cells in the low-,medium-,and high-dose Shenqi Dihuang Decoction groups presented tight arrangement,an increased cell count,improved morphology from a spindle-fiber shape to a cobblestone shape,and improved morphology and structure of mitochondrial membrane and cristae,compared with those in the high glucose group.Meanwhile,all the doses of Shenqi Dihuang Decoction inhibited ROS elevation to mitigate the peroxidation damage,lowered the Fe^(2+) and MDA levels and elevated the GSH level to inhibit lipid peroxidation,and activated the antioxidant pathway to upregulate the protein levels of Nrf2,HO-1,xCT,and GPX4.In conclusion,Shenqi Dihuang Decoction-medicated serum can inhibit high-glucose induced ferroptosis of HK-2 in vitro,which involves the antioxidant effect and the activation of the Nrf2/HO-1/GPX4 pathway.

关 键 词：糖尿病肾脏疾病 参芪地黄汤 铁死亡 Nrf2/HO-1/GPX4信号轴 

分 类 号：R285[医药卫生—中药学]