温脉通对血管平滑肌细胞增殖及相关因子表达的影响

Influences of Wenmaitong on multiplication of VSMC and expression of related factor

目的观察温脉通对血管紧张素Ⅱ(AngⅡ)诱导血管平滑肌细胞(VSMC)增殖的抑制作用及其对促VSMC生长因子的影响,以期探讨温脉通防治早期肢体动脉硬化性闭塞症(ASO)的作用机制。方法体外培养兔胸主动脉VSMC,以AngⅡ作为诱导因素,复制VSMC增殖模型,分别用温脉通全方及拆方药物血清进行干预。应用四唑盐(MTT)比色法测定各组VSMC增殖活性,用免疫细胞化学检测方法观察各组对血小板源性生长因子(PDGFBB)表达的影响。结果细胞经AngⅡ作用后其增殖活性明显增加,与空白组比较有显著性差异(P<0.01);而分别加入各组药物血清后,细胞增殖活性降低,与AngⅡ组比较有显著性差异(P<0.01),其中以大剂量组细胞增殖活性降低最为明显。血管平滑肌细胞PDGFBB免疫组化显色,空白组呈弱阳性表达(+/-);AngⅡ组呈强阳性表达(+++),与空白组比较有显著性差异;温脉通全方组PDGF-BB表达明显减弱(+),与空白组接近;温经益气拆方组和活血通脉拆方组PDGFBB呈中度表达(++),其阳性反应强度高于全方组,低于AngⅡ组。结论温脉通可呈剂量依赖性地抑制AngⅡ诱导的VSMC增殖、抑制PDGFBB的蛋白表达,提示温脉通防治ASO可能与抑制VSMC异常增殖及PDGF-BB的蛋白表达有关。

Objective To observe the inhibition effect of Wenmaitong on the multiplication of vascular smooth muscle cell (VSMC) induced by angiotension II (Ang II) and its influence on growth factor, and discuss its mechanism for preventing and treating early arteriosclerosis obliteration (ASO) of limbs. Methods To segregate media from rabbit aorta thoracalis for VSMC culture in vitro, and select 3^(rd) to 5^(th) generation cells to copy VSMC multiplication model induced by Ang II. The model was interfered by the serum with Wenmaitong or its different ingredients. The colorimetry of MTT was used to determine the VSMC multiplication activities and immunohistochemistry was used to observe the expressions of platelet-derived growth factor (PDGF-BB) in different groups. Results The multiplication activity of VSMC was obviously increased after Ang II action with a significant difference (P<0.01) compared with the blank group. After the Wenmaitong serum using, however, the multiplication activities in different groups were decreased. There was a significant difference (P<0.01) compared with the Ang II group. The decrease of VSMC multiplication activity in the high-dose group was the most obvious. VSMC PDGF-BB colorated after determined by immunohistochemistry. The intensity of positive reaction in the blank group was weak positive expression (+/-), and in the Ang II group, strong positive expression (+++) with a significant difference compared with the blank group. The expression of PDGF-BB in the Wenmaitong group was obviously decreased (+), approaching the blank group, and that in the ingredient group 1 and the ingredient group 2 was middle expression (++) and their intensities of positive reaction were higher than the Wenmaitong group and lower than the Ang II group. Conclusion Wenmaitong, with dose-dependability, can inhibit the VSMC multiplication activity induced by Ang II and the expression of PDGF-BB. The preventing and curatire effect of Wenmaitong on ASO may be related to its inhibition effect on VSMC abnormal multiplication and protein expression of PDGF-BB.