益气活血化痰法抗细胞凋亡防治动脉粥样硬化斑块形成的实验研究

Study on the Prevention and Treatment of Atherosclerosis Plaque by Yiqi Huoxue Huatan

目的：观察益气活血化痰法治疗载脂蛋白E基因敲除（apolipoprotein E-knockout ApoE-/-）小鼠动脉粥样硬化的作用机制.方法：选取健康6周龄雄性ApoE-/-小鼠30只,常规喂养1周后随机分为高脂饮食组、高脂＋参红颗粒组和普通饮食对照组,每组10只.前两组给予高脂饮食,中药干预组采用益气活血化痰中药参红通络颗粒进行干预.喂养4周后,三组小鼠分别给予0.9％生理盐水、参红通络颗粒（10.12g· kg-1/d）和0.9％生理盐水灌胃8周.各组3个月后分别抽取眼球血,采用ELISA法检测血清总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）值,取主动脉弓组织做HE染色观察动脉粥样硬化程度和病理组织学变化,免疫组化染色法检测凋亡相关基因Caspase-3蛋白的表达,TUNEL法检测斑块内细胞凋亡情况.结果：与高脂饮食组相比较,中药灌注组血脂水平明显降低,斑块面积减小并且斑块内细胞凋亡数量减少,明显降低细胞凋亡相关Caspase-3蛋白的表达.结论：上述研究结果表明,益气活血化痰法能通过死亡受体途径抑制斑块内细胞凋亡控制动脉粥样硬化病变进展.

OBJECTIVE： To observe the function and mechanism of Yiqi Huoxue Huatan therapy in atherosclerosis of apolipoprotein E （ApoE） -knockout mice. METHODS： 30 healthy ApoE-/- mice aged 6 weeks were selected. After 1 week conventional feeding the mice were randomly divided into high fat diet group, high fat diet ＋ Shenhong Tongluo granule group and normal diet control group, 10 rats in each group. The first two groups received high fat diet and the traditional Chinese medicine intervention group was treated by Shenhong Tongluo granule. After feeding for 4 weeks, 0. 9% saline, Shenhong Tongluo granule （ 10. 12g · kg-1/d） and 0. 9% normal saline were respectively given for 8 weeks in the three groups. For each group after 3 months the blood of eyeball was extracted and ELISA method was used to detect the TC, TG, HDL - C and LDL - C value. For the arch of aorta tissue gotten, HE staining was used to observe the extent of atherosclerosis and histopathological changes, and immunohistochemistry staining method was used to detect the apoptosis related to gene expression of caspase -3 protein expression. Apoptosis in plaque was detected by TUNEL method. RESULTS： Compared with the high fat diet group, in traditional Chinese medicine infusion group the blood lipid level, the area of atherosclerotic plaque, the numbers of apoptotic cells and the expression of Caspase - 3 protein were all significantly decreased. CONCLUSION ： Yiqi Huoxue Huatan can inhibite the progression of cell apoptosis in atherosclerosis plaque by death receptor pathway.