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

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(10-7 mol/L)、SPI 1组(ALD+SPI 10-7 mol/L)、SPI 2组(ALD+SPI 10-8 mol/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11 β-羟类固醇脱氢酶2 (11β-HSD 2)的mRNA表达.结果 ① MCs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.③ SPI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NFκB mRNA表达量呈显著正相关(P<001),NF-κB mRNA和MCP1 mRNA表达量也呈显著正相关(P<001).结论 可在一定程度上抑制ALD诱导的MCs氧化应激,减少NFκ-B和MCP-1的表达,该作用可能与其肾脏保护作用部分有关.%To observe the effects of spironolactone on aldosterone mediated oxidative stress and nuclear factor-κB( NF-κb ) expression in cultured rat glomerular mesangial cells( MCs ), and explore its mechanisms of re-no-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 mol/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-Kb, MCP-1, aldosterone synthase CYP11B 2, mineralocorticoid receptor ( MR) and 11 β-hydroxysteroid dehydrogenase ( 11 0-HSD 2 ) were detected by RT-PCR. Results ① CYP11B 2, MR and 11 0-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 Spironolactone, in some extent, can inhibit the expressions of NF-Kb, MCP-1 and oxidative stress induced by aldosterone in MCs, which may contribute to its partly reno-protection.