曲格列汀诱导高糖条件下肾系膜细胞表达HO-1  

Trelagliptin induces human renal mesangial cell expression heme oxyge-nase-1 under high glucose condition

作　　者：伍先明[1] 李爱琼 周丽丽[2] 彭小珊 李坚 WU Xian-ming;LI Ai-qiong;ZHOU Li-li;PENG Xiao-shan;LI Jian(Department of Geriatrics,The First People’s Hospital of Chenzhou,Chenzhou 423000,China;Children’s Hospital,The First People’s Hospital of Chenzhou,Chenzhou 423000,China;Chongqing Youth Vocational&Technical College,Chongqing 400712,China;Department of Basic Medicine,Xiangnan College of Medicine,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院老年病科,湖南郴州423000 [2]郴州市第一人民医院儿童医院,湖南郴州423000 [3]重庆青年职业技术学院,重庆400712 [4]湘南学院基础医学院,湖南郴州423000

出　　处：《中国病理生理杂志》2019年第12期2163-2168,共6页Chinese Journal of Pathophysiology

基　　金：湖南省教育厅科学研究项目(No.17C1475)

摘　　要：目的:观察曲格列汀在高糖条件下对人肾系膜细胞(HRMCs)表达血红素氧合酶1(HO-1)的影响,并初步探讨其分子机制。方法:在正常浓度(5 mmol/L)葡萄糖及高糖(25 mmol/L)条件下培养HRMCs,分别加入1、5和10μmol/L曲格列汀作用8~24 h。随后用RT-qPCR以及Western blot分别检测HO-1的mRNA和蛋白表达,比色法测定HO-1酶活性。此外,采用荧光探针检测细胞内活性氧簇(ROS)的变化,Western blot检测Akt磷酸化水平和核转录因子E2相关因子2(Nrf2)的表达水平。结果:高糖条件下HRMCs中HO-1的表达以及酶活性水平轻微增高,而经1、5和10μmol/L曲格列汀作用后,HO-1的mRNA、蛋白表达以及酶活性显著增高(P<0.05);同时,曲格列汀也能增加高糖条件下HRMCs的ROS水平并诱导Akt磷酸化,采用ROS清除剂NAC以及PI3K/Akt抑制剂LY294002处理后,HO-1的表达水平明显降低;此外,曲格列汀也可增强HRMCs核内转录因子Nrf2的表达,且NAC和LY294002能抑制曲格列汀诱导的Nrf2的表达。而沉默Nrf2后,曲格列汀诱导的HO-1表达减少。结论:曲格列汀可能能通过ROS和PI3K/Akt途径激活Nrf2而增加高糖条件下HRMCs表达HO-1。AIM:To observe the effect of trelagliptin on the expression of heme oxygenase-1(HO-1)in human renal mesangial cells(HRMCs)under high glucose condition and further to explore its molecular mechanism.METHODS:HRMCs were cultured at normal concentration(5 mmol/L)glucose and high concentration(25 mmol/L)glucose,and then were treated with trelagliptin at 1,5 and 10μmol/L for 8~24 h.The expression of HO-1 at mRNA and protein levels was determined by RT-qPCR and Western blot,and the activity of HO-1 enzyme was measured by colorimetry.The changes of reactive oxygen species(ROS)were detected by fluorescence probe,and the protein levels of phosphorylated Akt and nuclear factor E2-related factor 2(Nrf2)were determined by Western blot.RESULTS:The expression and enzyme activity of HO-1 in HRMCs were increased slightly under high glucose condition,while the expression at mRNA and protein levels and enzyme activity of HO-1 were increased significantly after treatment of trelagliptin at 1,5 and 10μmol/L.At the same time,trelagliptin also increased the level of ROS and induced Akt phosphorylation in HRMCs under high glucose condition.After treatment with NAC(ROS scavenger)and LY294002(PI3K/Akt inhibitor),the expression of HO-1 was significantly decreased.In addition,trelagliptin also increased the level of nuclear transcription factor Nrf2,and NAC and LY294002 inhibited the activation of Nrf2 induced by trelagliptin.After knock-down of Nrf2 expression,trelagliptin lost the ability to induce HO-1 expression.CONCLUSION:Trelagliptin may activate Nrf2 via ROS and PI3K/Akt pathway,and increase HO-1 expression in HRMCs under high glucose condition.

关 键 词：曲格列汀 氧化应激 人肾系膜细胞 PI3K/AKT信号通路 

分 类 号：R363.2[医药卫生—病理学] R587.2[医药卫生—基础医学]