螺内酯对高糖诱导的肾小球系膜细胞氧化应激的影响  

Effects of spironolactone on high glucose mediated oxidative stress in mesangial cells

作　　者：李庆[1] 张红萍[1] 

机构地区：[1]江苏建康职业学院信息中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2012年第12期1670-1674,共5页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的:观察螺内酯对高糖诱导的肾小球系膜细胞氧化应激的影响,并且探讨该作用是否与MAPKs家族信号通路相关。方法:采用高糖和(或)螺内酯处理人肾小球系膜细胞株(HMC)后,DHE荧光染色检测细胞内活性氧(ROS)水平,NBT试剂盒法检测细胞内超氧化物歧化酶(SOD)活性,Western blot法检测MAPKs家族ERK、JNK、p38MAPK的磷酸化及总蛋白水平。结果:DHE染色结果显示,高糖可明显增加HMC内ROS含量,而给予螺内酯干预处理后,细胞内ROS水平明显降低,与高糖刺激组相比,螺内酯干预组细胞内SOD活性明显增加(P<0.05);Western blot结果表明,高糖可明显增加HMC细胞内ERK及p38MAPK的磷酸化水平,给予螺内酯处理后可逆转高糖的上述作用(P<0.05),而各组之间磷酸化JNK及总ERK、p38MAPK、JNK蛋白水平无明显变化。结论:螺内酯可能通过下调MAPK家族信号蛋白的磷酸化水平,降低细胞内ROS含量,增加SOD活性,拮抗由高糖所引起的氧化应激,从而起到保护肾小球系膜细胞,延缓肾小球硬化的作用。Objective：To observe the effect of spironolactone on oxidative stress of glomerular mesangial cells induced by high glucose,and explore whether the MAPKs family signaling pathway was involved in this effect.Methods：DHE fluorescence staining was used to evaluate the intracellular production of reactive oxygen species（ROS）,NBT kit was used to detect the intracellular SOD activity,and Western blot was applied to identify the phosphorylated and total protein expression of MAPKs family genes,such as ERK,JNK and p38MAPK in cultured human mesangial cells（HMC） treated with high glucose or spironolactone.Results：DHE fluorescence staining showed that the production of ROS in HMC was markedly increased with the treatment of high glucose,while which was significantly decreased in the stimulation of spironolactone.Compared to the treatment of high glucose,intracellular SOD activity was significantly increased（P 0.05） with the treatment of spironolactone.Western blot analysis suggested that the high sugar significantly increased the expression of phosphorylation of ERK and p38MAPK in HMC cells,while given the treatment of spironolactone,these effects were reversed（P 0.05）.Furthermore,there were no remarkable differences among the groups in phosphorylated JNK,total protein level of ERK,p38MAPK,and JNK.Conclusion：Spironolactone may play a role in reducing the production of intracellular ROS and increasing the activity of SOD and antagonism of oxidative stress induced by high glucose via down-regulation of the phosphorylation level of MAPK family signaling.All these actions may contribute to the protection of the glomerular mesangial cells and postponement of kidney glomerulus sclerosis.

关 键 词：螺内酯 高糖 MAPK 肾小球系膜细胞 

分 类 号：R967[医药卫生—药理学]