温肾化痰方对ApoE基因敲除小鼠动脉粥样硬化易损斑块模型Orai1、Caspase、Bax、Bcl-2等基因及其相关蛋白表达的影响

Effect of Wenshen Huatan Decoction on the Expression of Orai1,Caspase,Bax and Bcl-2 Genes and Related Proteins in Aulnerable Plaque Model of ApoE Knockout Mice

目的观察温肾化痰方对ApoE基因敲除小鼠动脉粥样硬化易损斑块模型Orai1和Caspase3、Caspase9、Bax及Bcl-2等基因及其相关蛋白表达的影响。方法建立ApoE基因敲除小鼠动脉粥样硬化易损斑块模型,随机分成空白对照组、易损斑块组、阳性对照组、温肾化痰方低、中、高剂量组。13周后麻醉处死小鼠,采用RT-PCR技术检测主动脉平滑肌细胞Orai1、Caspase3、Caspase9、Bax及Bcl-2等基因的表达情况,并采用Western blot法检测主动脉窦Orai1、Caspase3、Caspase9、Bax及Bcl-2等蛋白表达变化。结果与空白对照组相比,易损斑块组Orai1、Caspase3、Caspase9、Bax mRNA及蛋白表达水平明显上升(P<0.01),而Bcl-2 mRNA及蛋白明显下降(P<0.01)。与易损斑块组相比,阳性对照组与温肾化痰方各剂量组均能下调上述基因(不包括Bcl-2)mRNA及蛋白表达水平,以及同时能上调Bcl-2的mRNA及蛋白表达,且中药剂量越高,调节效果越明显。与阳性对照组相比,温肾化痰方高剂量组下调Orail、Caspase3、Caspase9、Bax mRNA及蛋白水平以及上调Bcl-2 mRNA及蛋白水平的作用基本一致。这两组小鼠的上述基因及蛋白表达与空白对照组相比,统计无显著性差异。结论温肾化痰方能对应调节相关基因及蛋白表达从而干预易损斑块的形成及进展,进而起到保护心血管的作用。

Objective To observe the effects of Wenshen Huatan decoction on the expression of Orai1,Caspase3,Caspase9,Bax and Bcl-2 genes and their related proteins in atherosclerotic vulnerable plaque models of ApoE knockout mice.Methods The model of atherosclerotic vulnerable plaque in ApoE knockout mice was established and randomly divided into six groups,like blank control group,vulnerable plaque group,positive control group,Wenshen Huatan decoction low,medium and high dose groups.After 13 weeks,the mice were sacrificed by anesthesia.The expressions of Oria1,Caspase3,Caspase9,Bax and Bcl-2 genes in aortic smooth muscle cells were detected by RT-PCR technology.The expressions of protein associated with the above genes in the aortic sinus were also detected by Western blot technology.Results Compared with blank control group,the expression levels of Orai1,Casepase3,Casepase9 and Bax mRNA and protein in the vulnerable plaque group increased significantly(P<0.01),while the ones of Bcl-2 decreased significantly(P<0.01).Compared with the vulnerable plaque group,the mRNA and protein expression of the above genes(excluding Bcl-2)in the positive control group and Wenshen Huatan decoction group were down-regulated,and simultaneously the expression of Bcl-2’s mRNA and protein were up regulated.It’s also could be observed that the higher the dosage of the decoction,the more obvious the adjustment effect.Compared with the positive control group,the expression of those genes and protein in the high-dose group of Wenshen Huatan decoction were almost in the same.Among these two groups and blank control group,there was no statistically significant difference in the expression of those genes and proteins.Conclusion Wenshen Huatan decoction can prevent the formation and progression of vulnerable plaques by regulating the expression of relational genes and proteins,and thus protect the cardiovascular system.