当归芍药散对动脉粥样硬化小鼠的干预作用及DNA甲基转移酶1和PPAR-γ表达的影响

Effects of Danggui Shaoyao powder on the DNMT1 and the expression of PPARγ in atherosclerotic mice

目的研究当归芍药散对载脂蛋白E基因敲除(ApoE^(-/-))小鼠主动脉粥样硬化(atherosclerosis,As)斑块的干预作用及DNA甲基转移酶1(DNMT1)和PPAR-γ表达的影响。方法将30只8周龄雄性ApoE^(-/-)小鼠西方类型膳食喂养9周,随机分为模型组、立普妥组(阳性对照组)和当归芍药散组(n=10),药物干预9周后,检测血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,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)、血清DNA甲基化水平及DNMTs水平。行苏木素-伊红染色(hematoxylin and eosin staining,HE),采用IPP图像分析软件测量小鼠主动脉As斑块面积。免疫组化法检测各组小鼠主动脉As斑块内DNA甲基化转移酶1(DNMT1)和过氧化物酶体增殖物活化受体γ(PPARγ)的表达。结果与模型组相比,当归芍药散组和立普妥组小鼠血清TC、TG水平明显降低(P<0.05),当归芍药散组小鼠血清HDL-C水平明显升高(P<0.05)。与模型组相比,当归芍药散组小鼠主动脉As斑块面积显著降低(P<0.01),当归芍药散组小鼠主动脉斑块内DNMT1表达显著降低(P<0.05),PPARγ表达显著升高(P<0.05)。结论当归芍药散可改善As小鼠的血脂水平,减少As斑块面积,具有明确的抗As作用,其机制可能与抑制As小鼠血清甲基化水平和DNMTs水平,促进As小鼠主动脉斑块内PPARγ表达和抑制斑块内DNMT1表达有关。

Objective To study the effects of Danggui Shaoyao powder on the expression of DNA methyltransferase 1（ DNMT1） and PPARγ in apolipoprotein E knockout（ ApoE^-/-） mice with atherosclerosis. Methods Thirty 8 week-old male ApoE^-/- mice were fed with high fat diet for 9 weeks,and the mice were randomly divided into the control group,the Lipitor（ positive control group）,the Danggui Shaoyao powder group（ n = 10）. The levels of total cholesterol（ TC）,triglycerides（ TG）,low density lipoprotein cholesterol（ LDL-C）,high density lipoprotein cholesterol（ HDL-C） in serum were detected.The level of DNA Methylation and the DNMTs in serum were detected. HE staining was used to measure the area of atherosclerosis plaque of mice. Immuno-histochemical staining and image analysis were used to measure the expressions of DNMT1 and PPARγ in the atherosclerosis plaques of mice. Results Compared with the model group,the TC and TG levels of Danggui Shaoyao powder group and Lipitor group were significantly decreased（ P〈0. 05）,the HDL-C levels of Danggui Shaoyao powder group were significantly increased（ P〈0. 05）. Compared with the model group,the aorta As plaque area was significantly decreased in Danggui Shaoyao powder group（ P〈0. 05）. The expressions of DNMT1 of the Danggui Shaoyao powder group were significantly reduced（ P〈0. 05） while the expressions of PPARγ were significantly increased（ P〈0. 05）. Conclusion Danggui Shaoyao powder can improve the blood lipid level and the plaque area of As mice. The mechanism may be related to the reduction of DNA methylation level and the DNMTs level in serum,and also related to the inhibition of DNMT1 expression,and the promotion of PPARγ expression in mice with As.