螺内酯对醛固酮诱导的大鼠肾小球系膜细胞氧化应激及NF-κB、MCP-1表达的影响  

Effects of spironolactone on aldosterone mediated oxidative stress and NF-κB,MCP-1 expression in cultured rat glomerular mesangial cells

作　　者：刘皆[1] 叶山东[1] 胡圆圆[1] 徐将[1] 董崇周[1] 

机构地区：[1]安徽医科大学附属省立医院内分泌科,合肥230001

出　　处：《安徽医科大学学报》2012年第1期15-19,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省国际科技合作计划资助项目(编号:09080703041)

摘　　要：目的观察螺内酯(SPI)对醛固酮(ALD)诱导的大鼠肾小球系膜细胞(MCs)氧化应激和核因子-κB(NF-κB)、单核细胞趋化蛋白-1(MCP-1)表达的影响,探讨其肾脏保护机制。方法体外培养的MCs随机分为正常对照组(NG组)、ALD组(10-7mol/L)、SPI 1组(ALD+SPI 10-7mol/L)、SPI2组(ALD+SPI 10-8mol/L)和SPI 3组(ALD+SPI 10-9mol/L)。以流式细胞仪法检测MCs内活性氧(ROS)水平。RT-PCR法检测NF-κB、MCP-1、醛固酮合成酶(CYP11B 2)、醛固酮受体(MR)和11β-羟类固醇脱氢酶2(11β-HSD 2)的mRNA表达。结果①MCs表达CYP11B 2、MR和11β-HSD 2 mRNA。②与NG组比较,ALD刺激MCs 48 h后,细胞内ROS水平、NF-κB及MCP-1 mRNA表达明显增加。③SPI干预48 h后,与ALD组相比较,SPI 1组、SPI 2组、SPI 3组细胞内ROS水平、NF-κB及MCP-1 mRNA表达明显减少,且呈一定的剂量依赖性。④相关分析显示MCs内ROS水平与NF-κB mRNA表达量呈显著正相关(P<0.01),NF-κBmRNA和MCP-1 mRNA表达量也呈显著正相关(P<0.01)。结论 SPI可在一定程度上抑制ALD诱导的MCs氧化应激,减少NF-κB和MCP-1的表达,该作用可能与其肾脏保护作用部分有关。Objective To observe the effects of spironolactone on aldosterone mediated oxidative stress and nuclear factor-κB（NF-κB） expression in eultured rat glomerular mesangial cells（MCs）, and explore its mechanisms of reno-protection. Methods Rat MCs cultured with normal glucose concentration （5.6 mmol/L） , aldosterone （10-7 mol/L） and spironolactone（10-7 ,10-8 ,10-9 tool/L）,which assigned into group NG, group ALD, group SPI 1, SPI 2 and SPI 3. The levels of intracellular reactive oxygen species （ROS） were measured by flow cytometry, The mRNA expression of NF-κB, MCP-1, aldosterone synthase CYP11B 2, mineraloeortieoid receptor （MR） and 11 β- hydroxysteroid dehydrogenase （11 β-HSD 2） were detected by RT-PCR. Results ① CYP11B 2, MR and 11 β- HSD 2 mRNA expressions were detected in MCs. ② After treated by ALD for 48 h, the levels of ROS and the expression of NF-KB, MCP-1 mRNA were all increased in group ALD compared to group NG. ③ Compared with group ALD, the levels of ROS and the expression of NF-κB, MCP-1 mRNA in group SPI 1, SPI 2 and SPI 3 decreased significantly after treated by spironolactone for 48 h, with concentration-dependent. ④ There were significant positive correlation among ROS, NF-KB and MCP-1 （P 〈 0.01 ）. Conclusion 5pironolactone,in some extent,can inhibit the expressions of NF-KB, MCP-1 and oxidative stress induced by aldosterone Key words

关 键 词：氧化应激 核因子-κB 单核细胞趋化蛋白-1 螺内酯 醛固酮 糖尿病肾病 

分 类 号：R587.2[医药卫生—内分泌] R983.1[医药卫生—内科学]