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目的研究高同型半胱氨酸(Hhcy)型动脉粥样硬化(AS)大鼠主动脉含半胱氨酸天冬氨酸水解酶3(Caspase 3)、Bax及Bcl-2的表达。方法 20只SD大鼠随机分为实验组及对照组,实验组大鼠灌胃给予1 g·kg-1·d-1蛋氨酸复制Hhcy型AS大鼠模型,对照组大鼠给予等量生理盐水灌胃。饲养4个月后,处死大鼠。采用全自动分析仪检测血清同型半胱氨酸(Hcy)含量,HE染色检测主动脉斑块面积,免疫组化检测主动脉组织α-平滑肌肌动蛋白(α-SM-actin)表达,Western blot及逆转录聚合酶链反应(RT-PCR)法分别检测主动脉组织中Caspase 3、Bax及Bcl-2蛋白及其mRNA的表达。结果与对照组比较,实验组血清中Hcy含量显著提高,主动脉斑块面积及α-SM-actin阳性表达率增加(P均<0.01),表明Hhcy型AS大鼠模型复制成功。与对照组比较,实验组Bcl-2蛋白及mRNA表达上调,Bax及Caspase 3蛋白及mRNA表达下调(P均<0.01)。结论蛋氨酸诱导的Hhcy型AS大鼠通过调节Caspase 3、Bax及Bcl-2表达影响其主动脉硬化进程。
Objective To investigate the expression of Caspase 3, Bax and Bcl-2 in the aorta of rats with high homocysteine (Hhcy) type atherosclerosis (AS). Methods Twenty SD rats were randomly divided into experimental group and control group. The rats in experimental group were given Hg-AS model by intragastric administration of methionine (1 g · kg-1 · d-1), while rats in control group were given equal volume of normal saline Gavage. Four months after feeding, the rats were sacrificed. Serum levels of homocysteine (Hcy) were detected by automatic analyzer. Aortic plaque area was detected by HE staining. Α-SM-actin expression in aortic tissue was detected by immunohistochemistry. Western blot And reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of Caspase 3, Bax and Bcl-2 protein and their mRNA in aortic tissue respectively. Results Compared with the control group, the serum Hcy level in the experimental group increased significantly, and the area of aortic plaque and the positive rate of α-SM-actin increased (all P <0.01), indicating that Hhcy AS rat model was successfully replicated. Compared with the control group, the expression of Bcl-2 protein and mRNA were up-regulated in the experimental group, while the expressions of Bax and Caspase 3 protein and mRNA were down-regulated (all P <0.01). Conclusion Methionine-induced Hhcy AS rats affect the progression of aortic atherosclerosis by regulating the expressions of Caspase 3, Bax and Bcl-2.