白藜芦醇对血管紧张素Ⅱ诱导的成纤维细胞增殖的作用及其机制

Effect of resveratrol on the proliferation of NIH/3T3 induced by AngⅡ and related mechanism

目的研究白藜芦醇(Resv)对血管紧张素Ⅱ(AngⅡ)诱导成纤维细胞增殖的作用和机制。方法体外培养小鼠胚胎成纤维细胞(NIH/3T3),建立AngⅡ诱导的成纤维细胞增殖模型。采用细胞增殖-毒性检测试剂盒(CCK-8)检测细胞活力,Western blot技术检测Ⅰ型胶原蛋白(COLⅠ)、Ⅲ型胶原蛋白(COLⅢ)、基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶抑制因子-1(TIMP-1)的表达。结果与Control组相比,AngⅡ组NIH/3T3增殖明显,COLⅠ、COLⅢ、MMP-1和TIMP-1蛋白表达均显著增加,且TIMP-1/MMP-1增大;与AngⅡ组相比,加入10、20、30、100μmol/L白藜芦醇(Resv)对AngⅡ引起的NIH/3T3增殖有抑制作用(后期实验选用Resv的浓度为20μmol/L),COLⅠ、COLⅢ、MMP-1和TIMP-1蛋白表达明显减少,且TIMP-1/MMP-1降低;与AngⅡ组相比,AngⅡ预处理组引起的变化趋势同Resv组,强度不如Resv,且两者具有协同作用。结论白藜芦醇和AngⅡ预处理均可抑制AngⅡ引起的成纤维细胞增殖和活化,前者效果优于后者,两者合用效果更佳,其保护作用的机制与减少胶原蛋白的分泌和下调MMP-1与TIMP-1的比值有关。

Objective To explore the effect of resveratrol （Resv） on the proliferation of NIH/ 3T3 induced by Ang Ⅱ and related mechanism. Methods The fibroblast proliferation model was established by culturing NIH/3T3 with Ang Ⅱ. Cell viability was detected by cell counting kit-8 （CCK-8） reagent kit. Western blot assay was used to detect the expression of collagen I , collagen m, MMP-1 and TIMP-1. Results Compared with the normal group, the cell prolif- eration was increased and the expression of collagen Ⅰ-Ⅲ and MMP-1, TIMP-1 were significant- ly increased in Ang Ⅱ group ; Compared with the Ang Ⅱ group, the expression of collagen I , collagen m, MMP-1 and TIMP-1 and TIMP-1/MMP-1 was down regulated in Resv groups （the concentration of Resv was 10, 20, 30, 100 μmol/L and 20 p.moL/L was used in the following study）. The trend of change induced by angiotensin Ⅱ preconditioning （APC） was similar with Resv group. Conclusion Resv and Ang Ⅱ could inhibit the proliferation and activation of myocardial fibroblasts induced by Ang Ⅱ , which mechanism is related to decreasing secretion of collagen and down regulating TIMP-1/ MMP-1.