瑞舒伐他汀对载脂蛋白E基因敲除动脉粥样硬化小鼠的作用研究

Study on the effect of Rosuvastatin on apolipoprotein E gene knockout mice with atherosclerosis

目的观察主流降脂药瑞舒伐他汀对载脂蛋白E(ApoE-/-)基因敲除动脉粥样硬化小鼠血管壁保护及斑块的稳定作用,探讨干预高脂血症、动脉粥样硬化,防治心血管疾病的可能机制。方法20只ApoE-/-小鼠随机分为模型组和瑞舒伐他汀组,每组各10只,另10只C57BL/6/J小鼠设为正常组。模型组和瑞舒伐他汀组小鼠于6周起西方膳食饲料饲养至20周龄,瑞舒伐他汀组自16周龄起每日予以瑞舒伐他汀(10 mg/kg)灌胃,并同时对正常组及模型组每日给予饮水灌胃,观察至20周龄取材。采用生化方法检测各组小鼠血脂及炎症指标变化,采用苏丹Ⅳ染色法以及Image J检测小鼠主动脉斑块沉积情况,通过小动物主动脉超声测得小鼠中膜厚度等结果,使用Von Kossa染色法检测主动脉弓钙化程度。结果与正常组比较,模型组小鼠的血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL)水平均升高,差异有统计学意义(P<0.01)。与模型组比较,瑞舒伐他汀组小鼠的血清TG水平下降,差异有统计学意义(P<0.01)。瑞舒伐他汀组小鼠的血清LDL、TC水平与模型组比较,差异无统计学意义(P>0.05)。与正常组比较,模型组小鼠血清的肿瘤坏死因子-α(TNF-α)、细胞间黏附分子(ICAM-1)、单核细胞趋化蛋白(MCP-1)水平均升高,差异有统计学意义(P<0.01)。瑞舒伐他汀组小鼠的血清TNF-α、ICAM-1、MCP-1水平与模型组比较,差异无统计学意义(P>0.05)。模型组小鼠的主动脉斑块沉积面积平均值达39.66%,瑞舒伐他汀组小鼠的主动脉斑块沉积面积平均值达25.87%;瑞舒伐他汀组与模型组小鼠的主动脉斑块沉积面积比较,差异有统计学意义(P<0.01)。主动脉超声结果显示,各组小鼠的主动脉中膜厚度比较,差异无统计学意义(F=0.8965,P=0.4302)。与模型组比较,瑞舒伐他汀组的斑块易损指数降低,差异有统计学意义(P<0.01)。与模型组比较,瑞舒伐他汀组的血管钙化面积降低,差异有统计学意义(P<0.01)。结论瑞舒伐他汀对ApoE-/-基因敲除动脉粥样硬化小鼠可改善其斑块易损性和血管钙化程度,并具有抗动脉粥样硬化斑块沉积作用。

Objective To observe the effect of the mainstream lipid-lowering drug Rosuvastatin on the vascular wall protection and plaque stabilization of apolipoprotein E(ApoE-/-)gene knockout mice with atherosclerosis,and to explore the possible mechanisms of intervention in hyperlipidemia,atherosclerosis,and prevention and treatment of cardiovascular diseases.Methods A total of 20 ApoE-/-mice were randomly divided into the model group and the Rosuvastatin group,with 10 mice in each group,and the another 10 C57BL/6/J mice were set as normal control group.Mice in the model group and Rosuvastatin group were fed on Western diets from 6 weeks to 20 weeks of age,and the Rosuvastatin group was given Rosuvastatin(10 mg/kg)by gavage every day from 16 weeks of age,and simultaneously the normal group and the model group were given drinking water gavage every day,and observations were taken until the age of 20 weeks.Biochemical methods were used to detect changes in blood lipids and inflammation indicators in each group of mice.SudanⅣstaining method and Image J were used to detect aortic plaque deposition,and the intima media thickness was measured by small animal aortic ultrasound.Von Kossa staining method was used to detect the degree of aortic arch calcification.Results Compared with the normal group,the serum total cholesterol(TC),triacylglycerol(TG),and low-density lipoprotein(LDL)levels of the model group increased,and the differences were statistically significant(P<0.01).Compared with the model group,the serum TG level of mice in the Rosuvastatin group decreased,and the difference was statistically significant(P<0.01).There were no statistically significant differences in serum LDL and TC levels between the Rosuvastatin group and the model group(P>0.05).Compared with the normal group,the serum levels of tumor necrosis factor-α(TNF-α),intercellular adhesion molecule(ICAM-1)and monocyte chemoattractant protein(MCP-1)in the model group increased,with statistically significant differences(P<0.01).Compared with the model group,the serum TNF-α,ICAM-1 and MCP-1 levels of the Rosuvastatin group were not statistically different(P>0.05).The average aortic plaque deposition area of mice in the model group was 39.66%,and the average aortic plaque deposition area of the mice in the Rosuvastatin group was 25.87%.There was statistically significant difference in the area of aortic plaque deposition between the Rosuvastatin group and the model group(P<0.01).The results of aortic ultrasound showed that there was no statistically significant difference in the intima media thickness of each group of mice(F=0.8965,P=0.4302).Compared with the model group,the plaque vulnerability index of the Rosuvastatin group decreased,and the difference was statistically significant(P<0.01).Compared with the model group,the vascular calcification area of the Rosuvastatin group reduced,and the difference was statistically significant(P<0.01).Conclusion Rosuvastatin can improve the vulnerability of plaque and the degree of vascular calcification in ApoE-/-gene knockout mice with atherosclerosis,and has an anti-atherosclerotic plaque deposition effect.