【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。更多还原

【Abstract】 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 10 μmol/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 10 μmol/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(PI3 K/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 PI3 K/Akt pathway, and increase HO-1 expression in HRMCs under high glucose condition.更多还原