葛根素对血管紧张素Ⅱ诱导大鼠血管平滑肌细胞的增殖及蛋白激酶-α和核转录因子-κB表达的影响

The role of puerarin on the proliferation of rat vascular smooth muscle cells and the expressions of PKC-α and NF-κB induced by angiotensin Ⅱ

目的:观察葛根素对血管紧张素Ⅱ(AngⅡ)诱导血管平滑肌细胞(VSMC)增殖及对蛋白激酶-α(PKC-α)和核转录因子(NF)-κB表达的影响,探讨其可能存在的抑制VSMC增殖的分子机制。方法:体外培养Wistar大鼠的VSMC,给予单纯1.5×10-3mol/L葛根素处理2 h(P组)、单纯10-8mol/L AngⅡ处理24 h(A组)和1.5×10-3mol/L葛根素预处理2 h后再加10-8mol/L AngⅡ处理24 h(P+A组)刺激后,四氮唑盐法检测增殖情况,采用Western blot的方法检测PKC-α和NF-κB的表达。结果:对照组、P组、A组、P+A组VSMC增殖的A值分别为0.178±0.017、0.198±0.028、0.373±0.017和0.286±0.024。与对照组比较,A组和P+A组差异有统计学意义(均P<0.05);A组和P+A组VSMC中PKCα-、NFκ-B的表达与对照组比较,均显著增高(均P<0.05);P+A组较A组均有明显下降(均P<0.05)。结论:一定浓度的葛根素可通过下调PKCα-和NF-κB的表达来抑制AngⅡ引起的VSMC的增殖。

Objective：To observe the role of puerarin on the proliferation of rat vascular smooth muscle cells and the expressions of PKC-α and NF-κB induced by angiotensin Ⅱ and investigated to investigate the mechanism of puerarin suppressed the proliferation of vascular smooth muscle cells. Method：The vascular smooth muscle cells in Wistar rats were cultured in vitro and stimulated with the different factors for 24 h. Cell proliferation was determined with mitochondrial dehydrogenase activty （MTT assay）. Western bloting was used to detect the expression level of PKC-α and NF-κB. Result： 1. After vascular smooth muscle cells stimulating by different factors： Compared with control group, cell proliferation of group A and group P＋A were significantly increased（P〈0.05,n= 4） ; Compared with Group A, cell proliferation of group P＋A was significantly decreased（P〈0.05, n= 4） ;2. After vascular smooth muscle cells stimulating by different factors, the expression level of PKC-α in different groups： Compared with control group, the expression level of PKC-α in Group A and group P＋A were significantly increased（P〈0.05 ,n=4） ; Compared with Group A, the expression level of PKC-α in group P＋A was significantly decreased（P〈0.05, n= 4）; 3. After vascular smooth muscle cells stimulating by different factors, the expression level of NF-κB in different groups： Compared with control group, the expression level of NF-κB in Group A and group P＋A were significantly increased（P^0.05,n=4） ; Compared with Group A, the expression of level of NF-κB in group P＋A was significantly decreased（P〈0.05,n= 4）. Conclusion： Our study indicated that the appropriate concentration angiotensin Ⅱ could induce the significant proliferation of vascular smooth muscle cells with the increased expression of PKC-α and NF-κB; the appropriate concentration puerarin may suppress the proliferation of vascular smooth muscle cells induced by angiotensin Ⅱ through downregulating the expression of PKC-α and NF-κB.