摘要
目的观察氟伐他汀对血管紧张素（Ang）Ⅱ诱导的大鼠肾小管上皮细胞（NRK-52E）内核因子（NF）KB活性的影响。方法将NRK-52E细胞分为（1）对照组；（2）不同浓度及时间AngⅡ组；（3）AngⅡ（10^-6mol／L）＋SB203580（10μmol／L）组；（4）AngⅡ（10^-6mol／L）＋不同浓度氟伐他汀（10^-7、10^-6、10^-5mol／L）组；（5）AngⅡ（10^-6mol／L）＋氟伐他汀（10^-5mol／L）＋甲羟戊酸（10^-4mol／L）组。电泳迁移率变动分析法（EMSA）检测NF—KB活性变化。Western印迹方法检测p38丝裂原活化蛋白激酶（p38MAPK）磷酸化水平。RT—PCR方法检测单核细胞趋化因子（MCP-1）mRNA表达。结果AngH呈剂量依赖性上调NF-KBDNA结合活性、p38MAPK的磷酸化水平以及MCP-1mRNA表达（P〈0．01）。AugⅡ（10^-6mol／L）刺激5min即可增加p38MAPK蛋白磷酸化水平（P〈0．01）。Ang1I刺激30min后NF—KB活性显著升高（P〈0．01），2h达高峰（P〈0．01）。p38MAPK特异性抑制剂SB203580可阻断AngⅡ对NF-KB的激活作用（P〈0．01）。氟伐他汀可呈剂量依赖性下调AngⅡ诱导的NRK-52E细胞内p38MAPK磷酸化和NF-KB活化，以及其下游趋化因子MCP-1表达（P〈0．05）。甲羟戊酸（10^-4mol／L）可逆转氟伐他汀的作用（P〈0．05）。结论氟伐他汀可能通过抑制p38MAPK信号转导通路，下调AngⅡ诱导的NRK-52E细胞内NF-KB的活化。甲羟戊酸可部分逆转氟伐他汀的作用。
Objective To investigate the effect of fluvastatin on activalion of nuclear factor kappa B （NF-KB）induced by angiotensin Ⅱ （Ang Ⅱ ） in rat kidney tubular epithelial cells （NRK-52E）. Methods NRK-52E cells were divided into （ 1 ）control group ; （ 2 ）Ang Ⅱgroups with different concentration and time;（3）Angll （10.6 mol/L）＋SB203580 （10 μmol/L）group;（4） Ang Ⅱ （ 10-6 mol/L） ＋different fluvastatin concentration （ 10-7, 10-6, 10-5 mol/L）groups; （5）Ang Ⅱ （10.6 mol/L） ＋fluvastatin （10.5 mol/L） ＋mevalonate （10-4 mol/L）group. Electrophoretic mobility shift assays （EMSA） was used to detect NF-KB activation. Phosphorylation of cellular p38 mitogen- activated protein kinase （p38MAPK） was determined by Western blot. Monocyte chemoattractant protein （MCP）-I mRNA was determined by RT-PCR. Results Ang Ⅱ stimulated the DNA- binding activity of NF-KB,phosphorylation of p38MAPK and up-regulated the expression of MCP-1 mRNA in cultured NRK-52E cells in a dose-dependent manner （P〈0.01）. Ang Ⅱ （106moll/L） induced a rapid （5 minutes） elevation of the p38MAPK phosphorylation. NF-KB DNA binding activity was increased at as early as 30 minutes（P〈0.01）, peaked at 2 hours after Ang Ⅱ treatment （P〈0.01）. This stimulatory effect of Ang Ⅱ on NF-KB was blocked by SB203580 （a specific inhibitor of p38MAPK） （P〈0.01）. Incubation of cells with fluvastatin significantly inhibited the Ang Ⅱ-induced NF-KB activation and expression of MCP-1 mRNA in dose-dependent manner （P〈 0.05）. Exogenous mevalonate （10-4 mol/L） prevented the effect of fluvastatin on NF-KB activation （P 〈0.05）. Conclusions Fluvastatin reduces Ang H-induced NF-KB activation via the p38MAPK pathway in NRK-52E cells. Such effect of flurastatin is partly through blocked by mevalonate.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2009年第2期134-138,共5页
Chinese Journal of Nephrology
