罗格列酮抑制血管紧张素Ⅱ诱导的肾小球系膜细胞增殖及细胞外基质表达

Rosiglitazone Inhibitory Effect on Mesangial Cell Proliferation and Extracellular Matrix Expression Induced by Angiotensin Ⅱ

目的探讨过氧化物酶体增殖物活化受体γ(PPARγ)激动剂对血管紧张素Ⅱ(AngⅡ)诱导的肾小球系膜细胞(MC)增殖的抑制作用。方法体外培养小鼠MC,应用不同水平的AngⅡ(1,10,100nmol/L)或应用抗氧化剂乙酰半胱氨酸(NAC)或PPARγ激动剂罗格列酮预处理后再用AngⅡ刺激,于实验终点收集细胞,应用3H-胸腺嘧啶(3H-TdR)掺入法、细胞计数及流式细胞术测定MC增殖和细胞周期变化;抽提细胞总RNA,应用实时定量PCR检测转化生长因子-β1(TGF-β1)、纤溶酶原激活物抑制剂(PAI-1)和纤维连接蛋白(FN)mRNA表达;应用荧光探针2,7-二氯二氢荧光素乙酰乙酸(DCFDA)检测细胞内活性氧(ROS)的变化。结果1.MC应用100nmol/LAngⅡ刺激后,其3H-TdR掺入量及细胞数分别是基础状态下的2.14倍和2.32倍;罗格列酮可呈剂量依赖性抑制AngⅡ诱导的MC增殖,其抑制率可达85%以上。2.MC应用100nmol/LAngⅡ刺激24h后,48%的细胞进入S期和G2/M期;罗格列酮呈剂量依赖性阻断AngⅡ诱导的细胞周期变化。3.罗格列酮可呈剂量依赖性抑制AngⅡ诱导的MCTGF-β1、PAI-1和FNmRNA表达。4.MC应用100nmol/LAngⅡ刺激60min后,ROS产生是对照组的3.85倍。罗格列酮可呈剂量依赖性抑制AngⅡ诱导的MCROS产生,10μmol/L罗格列酮几乎完全阻断AngⅡ诱导的ROS产生。结论PPARγ激动剂罗格列酮通过抑制氧化应激阻断AngⅡ诱导的MC增殖和细胞外基质表达。

Objective To investigate the inhibitory effect of peroxisome proliferator - activated receptor - γ （ PPARγ） agonist on mesangial cell（MC） proliferation and extracellular matrix expression induced by angiotensin Ⅱ （ AngⅡ ）. Methods The incorporation of 3 H - thymidine（ 3H- TdR） and cell count were used as the measurement of MC proliferation. MC cell -cycle was analyzed by flow cytometry. Mouse primary MC was treated with various concentration of Ang Ⅱ （ 1,10,100 nmol/L） in the presence or absence of N - acytosistin （ NAC ） or rosiglitazone. Transforming growth factor - β1 （ TGF -β1 ）, plasminogen activator inhibitor - 1 （PAI - 1 ）, and fibronectin （ FN ） mRNA expression were determined by real time - PCR. Reactive oxygen species （ROS） production was measured by 2,7 - dichlorofluorescein diacetate （DCFDA） fluorescence. Results 1. One hundred nmol/L Ang Ⅱ increased 3 H - TdR incorporation and cell number by 2.14 and 2.32 fold, respectively. Ang Ⅱ - induced MC proliferation was inhibited by PPARγ agonist rosiglitazone with dose - dependent manner in mouse MC. 2. One hundred nmol/L Ang Ⅱ stimulation for 24 h induced 48% MC processed to S and G2/M phase. Rosiglitazone significantly blocked Ang Ⅱ increased cell number in S and G2/M phase. 3. Rosiglitazone reduced Ang Ⅱ - induced TGF - β1 ,PAI - 1 ,and FN mRNA expression with dose - dependent manner. 4. One hundred nmol/L Ang U stimulation for 60 min increased ROS production by 3.85 folds. Rosiglitazone significantly inhibited Ang Ⅱ - induced ROS production. Ten μmol/L rosiglitazone almost completely blocked Ang Ⅱ - induced ROS production. Conclusion PPARγ agonist rosiglitazone could block Ang Ⅱ - induced MC proliferation and extracellular matrix expression via inhibition of ROS production.