氯沙坦抑制高糖及间歇性高糖诱导NRK-52E细胞凋亡研究  

EFFECT OF LOSARTAN ON HIGH GLUCOSE AND FLUCTUATING HIGH GLUCOSE INDUCED APOPTOSIS IN NRK-52E

作　　者：钟宇华 梁华晟 苏会璇 

机构地区：[1]广西医科大学第九附属医院内分泌科,北海536000

出　　处：《广西医科大学学报》2010年第5期669-672,共4页Journal of Guangxi Medical University

基　　金：广西科技厅自然科学基金资助项目(桂科自No.0991294);广西北海市科学研究与技术开发计划项目(北科合字200901059)

摘　　要：目的:探讨氯沙坦对高糖及间歇性高糖诱导大鼠肾小管上皮细胞株NRK-52E凋亡的抑制作用。方法:作用72h后,分别采用MTT检测NRK-52E细胞活力,细胞凋亡采用流式细胞仪检测Annexin Ⅴ-FITC,细胞ROS含量采用CM2H2DCFDA试剂盒检测。Western blot检测空白组(0 mmol/L)、正常葡萄糖组(5mmol/L)、高糖干预组(25 mmol/L)、间歇性高糖组(5～25 mmol/L交替)、氯沙坦组(10 μmol/L)、高糖氯沙坦组(10 μmol/L氯沙坦预处理后在高糖培养)、间歇性高糖氯沙坦组(10 μmol/L)中凋亡调控标记蛋白Caspase-3及Caspase-9在NRK-52E细胞表达。结果:高浓度葡萄糖及间歇性高糖均上调Caspase-3及Caspase-9表达,细胞活性降低、细胞凋亡率升高,ROS含量显著上升,其中在间歇性高糖作用更显著。应用氯沙坦干预后可以部分下调高糖或间歇性高糖中Caspase-3及Caspase-9表达,细胞凋亡率显著下降,ROS含量下降。结论:氯沙坦可通过抑制高糖及间歇性高糖诱导NRK-52E细胞氧化应激从而减轻细胞凋亡。Objective:To study whether losartan can protect NRK-52E from high glucose and fluctuating high glucose induced apoptosis.Methods:Western blot of Caspase-3 and Caspase-9 in control group (0 mmol/L glucose),normal glucose group(5 mmol/L glucose),high glucose group(25 mmol/L glucose),fluctuating high glucose group(5 mmol/L glucose/25 mmol/L glucose),losartan group(10 nmmol/L losartan),losartan and high glucose group(10 nmmol/L losartan and 25 mmol/L glucose),losartan and fluctuating high glucose group(10 nmmol/L losartan and 5 mmol/L glucose/25 mmol/L glucose) was performed.Then cell viability and apoptosis were used to measure MTT and Annexin Ⅴ,ROS was detected by CM2H2DCFDA after 72 h.Results:Losartan could increase cell viability and decrease cell apoptosis in high glucose or fluctuating high glucose.The expression of Caspase-3 and Caspase-9 was up-regulated by hyperglycemia and fluctuating high glucose.However losartan could partly down-regulate the expression of Caspase-3 and Caspase-9 under fluctuating high glucose high glucose condition or high glucose.Fluctuating high glucose could also increase more ROS than high glucose,and losartan could decrease ROS under fluctuating high glucose high glucose condition or high glucose.Conclusion:Losartan maybe protect NRK-52E from high glucose and fluctuating high glucose which induced oxidative stress,and decrease cell apoptosis.

关 键 词：氯沙坦 抑制作用 高糖诱导 间歇性高糖 细胞凋亡率 HIGH GLUCOSE Caspase-9 Caspase-3 流式细胞仪检测 细胞氧化应激 高浓度葡萄糖 含量 细胞表达 上皮细胞株 ROS 细胞活力 活性降低 凋亡调控 标记蛋白 预处理 

分 类 号：R[医药卫生]