平肝潜阳法对SHR血管平滑肌细胞增殖及HSP27表达的影响

Effects of the calming liver and suppressing yang method on proliferation and the expression of heat shock protein 27 in vascular smooth muscle cells of spontaneously hypertensive rats

目的探讨平肝潜阳法对体外培养的自发性高血压大鼠(SHRs)血管平滑肌细胞(VSMCs)增殖的影响,及与热休克蛋白27(HSP27)表达的关系。方法采用改良的贴块法行SHR主动脉VSMC的分离培养,α-actin单克隆抗体免疫组织化学染色鉴定培养细胞;以MTT法反映VSMC增殖情况;用RT-PCR和Western印迹方法检测HSP27基因和蛋白的表达水平。结果对照组和治疗组均能抑制VSMC增殖,两组有显著差异(P<0.01);RT-PCR和Western印迹分析显示,与正常组比较,模型组HSP27表达明显增强(P<0.01);与模型组比较,对照组和治疗组HSP27表达明显减弱(P<0.01)。结论平肝潜阳法通过抑制VSMC增殖可能与抑制HSP27表达有关,是改善高血压血管重构的重要作用分子机制之一。

Objective To explore the effects of the calming liver and suppressing yang method on vascular smooth muscle cells （VSMCs） proliferation in vitro and the expression of heat shock protein 27 （HSP27） in VSMCs of spontaneously hypertensive rats（SHRs）. Methods VSMCs of aorta in SHRs were cultured by the modified explant method. The immunohistochemical technique was used to identify the VSMCs. The effects of the calming liver and suppressing yang method on the proliferation of VSMCs were examined by means of Mq^I＂. The expression of HSP27 gene was detected by RT-PCR, and the expression of its protein was assessed by Western blot. Results Control and treatment groups had the inhibited proliferation of VSMCs, and both of them had significant difference （P 〈 0. 01 ）. Compared with con- trol group, model group had significantly increased protein and gene expressions of HSP27 （ P 〈 0.01 ）. Compared with model group, control and treatment groups had significantly decreased protein and gene expressions of HSP27 （ P 〈 0.01 ）. Conclusions The calming liver and suppressing yang method can significantly inhibit VSMCs proliferation, and this effect may be related to inhibiting the expression of HSP27. It is one of the molecule mechanisms of improving vascular remodeling in spontaneously hypertension.