丹酚酸B和丹参酮ⅡA对大鼠胸主动脉外膜成纤维细胞增殖的影响

Effect of salvianolic acid B and tanshinone Ⅱ A on the proliferation of rat thoracic aorta adventitial fibroblast

目的探讨丹酚酸B(salvianolic acid B,SAB)和丹参酮ⅡA(tanshinoneⅡA,TA)单独用药、二者配伍对SpragueDawley(SD)大鼠胸主动脉外膜成纤维细胞增殖的影响,以及在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导下,SAB和TA有效成分配伍对大鼠胸主动脉外膜成纤维细胞增殖的影响。方法采用四甲基偶氮唑盐(methyl thiazolyl tetrazoli-um,MTT)方法检测SAB和TA对大鼠胸主动脉成纤维细胞增殖的影响,流式细胞术检测SAB和TA有效成分配伍对细胞周期的影响。结果 MTT检测结果显示,SAB和TA单独用药均可不同程度抑制成纤维细胞增殖,通过一系列浓度筛选,分别获得3个有效浓度,将其两两组合后比较对细胞生长的抑制作用,结果显示TA(10^(-8)mol·L^(-1))和SAB(10-5mol·L^(-1))组合显示明显的抑制作用(P<0.01)。而且,TA(10^(-8)mol·L^(-1))和SAB(10^(-5)mol·L^(-1))配伍组合亦能对AngⅡ诱导的细胞增殖产生明显的抑制作用,进一步采用流式细胞术观察SAB、TA及二者配伍对细胞周期的影响,结果显示,SAB和TA配伍明显地阻滞细胞在G_0/G_1期。对AngⅡ预刺激的细胞,SAB和TA组合,也明显阻滞细胞在G_0/G_1期;而SAB和TA分别给药,细胞主要被阻滞在S期。结论SAB和TA对成纤维细胞的增殖有明显的抑制作用,对AngⅡ诱导的细胞增殖也有明显的抑制作用,并且这种抑制作用主要是通过将细胞阻滞在G_0/G_1期而发挥作用的。

Aim To explore the effects of salvianolic acid B（SAB） and tanshinone Ⅱ A（TA） alone or the compatibility of these two effective components on the proliferation of rat vascular adventitial fibroblasts,and to observe the effects of the compatibility of the two on cell proliferation with the stimuation of angiotensin Ⅱ（Ang Ⅱ）.Methods The effects of SAB and TA alone or the compatibility of the two on cell proliferation were detected by methyl thiazolyl tetrazolium（MTT）method,and flow cytometry was adopted to detect cell cycle with or without the induction of Ang Ⅱ.Results It was shown that SAB and TA alone could inhibit fibroblast proliferation in different degree.Through a series of concentration screening,three effective concentrations were obtained respectively and then the inhibition of cell growth was detected by Pairwise compatibilities.The results showed that the compatibility of TA（10^-8mol·L^-1） and SAB（10^-5mol·L^-1） had the most significant inhibitory effect（P〈0.01）,and they could inhibit cell proliferation,further flow cytometry was adopted to detect drug effects on cell cycle,the results indicated that the compatibility of SAB and TA mainly blocked the cells in G0/G1 phase.Induced by Ang Ⅱ,the compatibility of SAB and TA group,compared with Ang Ⅱ group,blocked thee cells in G0/G1phase;and compared with combination of SAB and TA group,it mainly blocked cell cycle in S phase.Conclusion SAB and TA have certain inhibitory effect on fibroblast proliferation,also they could inhibit the proliferation induced by Ang Ⅱ,mainly by blocking the cells in G0/G1 phase.