三七总皂苷对ApoE^(-/-)小鼠动脉粥样硬化斑块形成及自噬的影响

The Influence of Panax Notoginseng Sponins on Atherosclerotic Plaque Formation and Autophagy in ApoE^(-/-) Mice

目的:探讨三七总皂苷对载脂蛋白E基因敲除ApoE^(-/-)小鼠动脉粥样硬化(atherosclerosis,AS)斑块形成的影响及其作用机制。方法:8只雄性C57BL/6J小鼠为对照组,32只ApoE^(-/-)小鼠随机分为模型组、三七总皂苷低剂量组(40 mg·kg^(-1))、三七总皂苷中剂量组(80 mg·kg^(-1))、三七总皂苷高剂量组(160 mg·kg^(-1)),每组各8只。对照组给予普通饲料喂养,其余各组采用高脂饲料喂养的同时进行药物干预,对照组、模型组给予等量生理盐水灌胃,每日1次,共干预8周,观察小鼠体质量和精神状态。全自动生化仪检测小鼠血清中总胆固醇(total cholesterol,TC)、三酰甘油(triacylglycerol,TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)的水平;油红O染色观察主动脉斑块分布情况;HE染色观察小鼠主动脉病理变化;Western Blot检测小鼠主动脉中Beclin1蛋白表达水平;免疫荧光法检测LC3的表达水平;透射电镜观察小鼠主动脉自噬体形成情况。结果:与对照组比较,模型组小鼠体质量增加,血清TG、TC、LDL-C水平升高,主动脉内壁斑块增多,Beclin1、LC3蛋白表达水平升高,自噬体数量增多;与模型组比较,三七总皂苷可降低动脉粥样硬化小鼠体质量及血清中TG、TC、LDL-C水平,减少主动脉内壁斑块,降低Beclin1、LC3蛋白表达水平及自噬体数量,差异均具有统计学意义(P<0.05)。结论:三七总皂苷可抑制ApoE^(-/-)小鼠AS斑块的形成,其作用机制可能与调控自噬水平相关。

Objective:To probe into the influence of panax notoginseng saponins on atherosclerotic plaque formation in apolipoprotein E knockout ApoE^(-/-) mice and its mechanism.Methods:A total of 8 male C57BL/6J mice were selected into the control group,and 32 ApoE^(-/-) mice were randomly divided into the model group,the low dose group of panax notoginseng saponins(40 mg·kg^(-1)),the medium dose group of panax notoginseng saponins(80 mg·kg^(-1)),and the high dose group of panax notoginseng saponins(160 mg·kg^(-1)),with 8 cases in each group.The mice in the control group were fed with common feed,while the mice in the other groups were fed with high-fat feed combined with drug intervention.The control group and model group were given the same amount of normal saline by gavage,once a day for a total of 8 weeks,during which the body mass and mental state of the mice were observed.The total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C) and high density lipoprotein cholesterol(HDL-C) in the serum of mice were detected by automatic biochemical analyzer.The distribution of aortic plaque was observed by oil-red O staining.The pathological changes of mouse aorta were observed by HE staining.The expression of Beclin1 protein in aorta was detected byWestern blot.The expression level of LC3 was detected by immunofluorescence assay.The autophage of aorta was observed by transmission electron microscope.Results:Compared with the control group,the body mass of mice in the model group increased,the levels of serum TG,TC,LDL-C increased,the number of plaque in the inner wall of aorta increased,the expression levels of Beclin1 and LC3 protein increased significantly,and the number of autophages also increased.Compared with the model group,panax notoginseng saponins could reduce the body mass,TG,TC,LDL-C levels in serum,plaque in aortic inner wall,protein expression levels of Beclin1 and LC3,and the number of autophages in atherosclerosis mice.All the differences were statistically significant(P<0.05).Conclusion:Panax notoginseng saponins can inhibit the formation of AS plaque in ApoE^(-/-) mice,and its mechanism may be related to the regulation of autophagy.