过氧化物酶体增殖物活化受体γ激动剂抑制血管紧张素Ⅱ诱导的肾小球系膜细胞单核细胞趋化蛋白-1的表达

Peroxisome Proliferator-Activated Receptor-Agonist Inhibited Angiotensin Ⅱ Induced Monocyte Chemoattractant Protein-1 Expression in Human Mesangial Cells

目的探讨氧化应激对血管紧张素Ⅱ(AngⅡ)诱导的肾小球系膜细胞单核细胞趋化蛋白-1(MCP-1)表达的影响,观察过氧化物酶体增殖物活化受体γ(PPARγ)激动剂对AngⅡ诱导的MCP-1表达的抑制作用。方法体外培养人肾小球系膜细胞。分为正常对照组、不同水平AngⅡ(1、10、100nmol/L)刺激组及乙酰半胱氨酸(NAC,10μmol/L)和罗格列酮(10μmol/L)预处理30min后加入AngⅡ(100nmol/L)刺激组。应用实时定量聚合酶链反应(PCR)检测各组MCP-1的表达,荧光探针2′,7′-二氯二氢荧光素乙酰乙酸(DCFDA)检测细胞内活性氧的变化。结果1.AngⅡ呈剂量依赖性的诱导肾小球系膜细胞MCP-1表达,1、10和100nmol/LAngⅡ刺激12h后MCP-1表达是对照组的1.80、2.51和3.57倍。2.AngⅡ可呈剂量依赖性诱导肾小球系膜细胞活性氧(ROS)的产生,1、10和100nmol/LAngⅡ刺激60min,ROS产生分别是对照组的1.82、2.92和4.08倍。3.Losartan完全阻断了AngⅡ诱导的ROS产生,而PD123319对AngⅡ诱导的ROS产生无抑制作用。4.抗氧化剂NAC显著抑制AngⅡ诱导的肾小球系膜细胞MCP-1表达。5.PPARγ受体激动剂罗格列酮10μmol/L可显著抑制AngⅡ诱导的系膜细胞ROS产生和MCP-1表达。结论AngⅡ通过诱导氧化应激促进肾小球系膜细胞MCP-1表达;PPARγ激动剂通过抑制氧化应激阻断AngⅡ诱导的MCP-1表达。

Objective To explore the effect of oxidative stress on angiotensin Ⅱ （ AngⅡ） - induced monocyte chemoattractant protein - 1 （ MCP - 1 ） expression, and investigate the inhibitory effect of peroxisome proliferator - activated receptor -γ （PPARγ） agonist on Ang Ⅱ - induced MCP - 1 expression. Methods Human primary mesangial cells were cultured in vitro in various concentration of Ang Ⅱ （1, 10, 100 nmol/L） in the presence or absence of N - acytosistin （NAC, 10 μmol/L） or rosiglitazone （10 μmol/L）. MCP - 1 gene expression was determined by real time reverse transcription- polymerase chain reaction（ RT- PCR）. Reactive oxygen species （ROS） production was measured by 2′ ,7′ -dichlorofluorescein diacetate （DCFDA） fluorescence. Results 1. Ang Ⅱ induced MCP- 1 expression in dose - dependent manner in human mesangial cells. 1,10, and 100 nmol/L Ang Ⅱ increased MCP - 1 expression by 1.80, 2.51, and 3.57 fold. 2. Ang Ⅱ induced ROS production in dose -dependent manner. 1,10, and 100 nmol/L Ang Ⅱ increased ROS production by 1.82, 2.92 and 4.08 fold. 3. Ang Ⅱ receptor 1 （ AT1 ） receptor antagonist losartan almost completely blocked Ang Ⅱ - induced ROS production, however, AT2 receptor antagonist PD123319 was not affected by ROS production. 4. Anti - oxidant NAC significantly inhibited Ang Ⅱ - induced MCP- 1 expression in human mesangial cells. 5. PPARγ/ agonist rosiglitazone 10 μmol/L significantly inhibited Ang Ⅱ- induced ROS production and MCP - 1 expression in human mesangial cells. Conclusions Ang Ⅱ induced MCP - 1 expression in human mesangial cells via ROS production, PPARγ/ligand rosiglitazone may block Ang Ⅱ - induced MCP - 1 expression via inhibition of ROS production.