参红通络颗粒对ApoE^(-/-)小鼠动脉粥样硬化斑块内细胞凋亡的影响及其抗动脉粥样硬化作用机制

Influence of Shenhong Tongluo Granules in apoptosis in atherosclerosis plaques of ApoE^(-/-) mice and its mechanisms of anti-atherosclerosis

目的:探讨参红通络颗粒对载脂蛋白E基因缺陷(ApoE^(-/-))小鼠血脂水平、主动脉粥样硬化斑块及斑块内细胞凋亡的影响,阐明参红通络颗粒抗动脉粥样硬化的作用机制。方法:选取无特异病原(SPF)级6周龄雄性ApoE^(-/-)小鼠40只,适应性喂养1周后,按随机数字表法分为正常对照组、模型组、低剂量中药治疗组和高剂量中药治疗组,每组10只。正常对照组给予普通饲料,其余各组小鼠通过喂饲高脂饲料建立小鼠动脉粥样硬化模型,造模成功后,低和高剂量中药治疗组小鼠分别给予参红通络颗粒5.06和10.12g·kg^(-1)·d-1灌胃治疗6周。6周后腹腔麻醉下摘眼球取血,测定各组小鼠血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平;分离主动脉弓,行HE染色,观察各组小鼠主动脉粥样硬化斑块形态并计算斑块面积;TUNEL法检测小鼠主动脉粥样硬化斑块内细胞凋亡情况,计算凋亡指数;免疫组织化学染色法检测凋亡相关基因Bcl-2和Bax蛋白表达水平。结果:低剂量中药治疗组小鼠血清TG水平明显低于模型组(P<0.05),低和高剂量中药治疗组小鼠血清HDL-C水平明显高于模型组(P<0.05或P<0.01);高剂量中药治疗组小鼠血清TG和HDL-C水平均明显高于低剂量中药治疗组(P<0.05)。与模型组比较,低和高剂量中药治疗组小鼠主动脉粥样硬化斑块面积明显缩小(P<0.01),主动脉粥样斑块内细胞凋亡指数明显降低(P<0.01),主动脉粥样斑块内Bcl-2蛋白表达水平明显降低、Bax蛋白表达水平明显升高(P<0.05或P<0.01)。结论:参红通络颗粒能够显著升高ApoE^(-/-)小鼠血清HDL-C水平,并通过调控Bcl-2、Bax蛋白表达抑制细胞凋亡,从而有效抑制动脉粥样斑块进展。

Objective：To explore the influence of Shenhong Tongluo Granules in the lipid levels,atherosclerosis plaques and apoptosis in the plaques of atherosclerotic apolipoprotein E （ApoE）-knokout mice,and to clarify the mechanism of Shenhong Tongluo Granules in anti-atherosclerosis.Methods：Forty SPF male mice aged six-week old＆amp;nbsp;were randomly divided into normal control group,model group,low dose of traditional chinese medicine （TCM） group and high dose of TCM group （n=10）after fed adaptively for 1 week.The mice in normal control group were fed with normal diet continuously, the mice in other three groups were fed with high fat diet to establish atherosclerosis model,and then the mice in low and high doses of TCM groups were respectively given Shenhong Tongluo Granules （5.06 and 10.12 g·kg-1 · d-1 ）by gavage for 6 weeks. The levels of serum total cholesterol （TC）,triglycerides （TG）,low density lipoprotein cholesterol （LDL-C）and high density lipoprotein cholesterol （HDL-C）of the mice in various groups were measured after the eyeball blood was collected.The aortic arch tissue was separated,and the morphotogy and the areas of atherosclerotic plaques were observed and calculated by HE staining;the apoptosis in plaques of the mice was evaluated by TUNEL method, and the apoptosis index was calculated. The expressions of apoptosis-assoicated gene Bcl-2 and Bax protein were measured by immunohistochemical staining.Results：The serum level of TG in low dose of TCM group was lower than that in model group （P 〈0.05）;the serum levels of HDL-C in low and high doses of TCM groups were lower than that in model group （P 〈0.05 or P 〈0.01）;the serum levels of TG and HDL-C in high dose of TCM group was higher than that in low dose of TCM group （P 〈0.05）.The areas of atherosclerotic plaques of the mice in low and high doses of TCM groups were significantly smaller than that in model group （P 〈0.01）.The apoptosis index of mice in low and high doses of TCM groups were significantly lower than that in model group （P 〈0.01）;the expression levels of Bcl-2 protein in atherosclerosis plaques of the mice in low and high doses of TCM groups were lower than that in model group,while the expression levels of Bax were significantly higher than that in model group （P 〈0.05 or P 〈0.01）.Conclusion：Shenhong Tongluo Granules could effectively reduce the serum lipid level of ApoE-/-mice,meanwhile it could inhibit apoptosis by regulating the expressions of Bcl-2 and Bax protein,and then inhibit the progression of atherosclerotic plaques.