丹参酮ⅡA对ApoE^(-/-)动脉粥样硬化小鼠腹腔巨噬细胞胆固醇流出相关基因表达的影响

The Effect of Tanshinone ⅡA on Macrophages Reverse Cholesterol Transport Related Genes Expression in ApoE^(-/-) Mice

目的:观察丹参酮ⅡA对ApoE基因敲除小鼠主动脉粥样斑块形成及对腹腔巨噬细胞胆固醇逆向转运相关基因的影响。方法:20只Apo E-/-小鼠随机分成模型组、丹参酮ⅡA组,10只C57BL/6J小鼠作为空白对照组。模型组、丹参酮ⅡA组给予高脂饲料喂养,空白组给予普通饲料。丹参酮ⅡA组每日灌胃丹参酮ⅡA(30mg/kg),模型组、空白对照组灌胃予等量生理盐水。检测各组小鼠血清TG、TC、HDL-C、LDL-C水平。油红O染色法观察各组小鼠主动脉脂质沉积情况,HE染色观察小鼠主动脉结构变化及动脉粥样硬化程度,采用RT-PCR和Western blot法检测腹腔巨噬细胞ABCA1、ABCG1、CD36的m RNA和蛋白表达,流式细胞术检测巨噬细胞CD36含量。结果:模型组小鼠血清HDL-C水平显著低于空白对照组,血清TG、TC、LDL-C水平显著高于空白对照组;组织形态学显示,模型组小鼠主动脉管腔中明显可见粥样斑块形成,管壁沉积大量脂质,腹腔巨噬细胞ABCA1 m RNA和蛋白表达显著低于空白对照组,ABCG1表达变化不明显,CD36 m RNA和蛋白表达显著高于空白对照组。与模型组比较,丹参酮ⅡA组小鼠血脂水平及动脉损伤水平明显改善;丹参酮ⅡA组巨噬细胞ABCA1基因和蛋白表达显著高于模型组,CD36基因和蛋白表达显著低于模型组,两组ABCG1表达差异无统计学意义。结论:丹参酮ⅡA对Apo E-/-小鼠主动脉斑块形成具有良好的抑制的作用,其机制可能与调控ABCA1及CD36等与巨噬细胞胆固醇流出相关基因及血脂有关。

Objective： To observe the effect of Tanshinone Ⅱ A on the formation of aortic atherosclerotic plaque in ApoE-/-mice and on peritoneal macrophages reverse cholesterol transport related genes. Methods： The 20 ApoE-/- mice were randomly divided into model group and Tanshinone Ⅱ A group, and 10 C57BL/6J mice with the same age and genetic background were used as the blank control group. The mice were given high fat diet in model group and Tanshinone ⅡA group, while normal diet in the blank control group. The mice were given Tanshinone I A 30 mg/ kg by intragastrie administration once a day for 8 weeks in Tanshinone ⅡA group, and normal saline in the model group and blank control group. The levels of TG, TC, HDL-C and LDL-C in serum were detected by automatic biochemical analyzer. Aortic structure, atherosclerosis level and lipid deposition were examined by HE and oil red O staining method. The mRNA of ABCA1, ABCGI, CD36 wre tested by RT-PCR; the expression of ABCA1, ABCG1 protein were tested by Western blot method; flow eytometry was used to detect the level of CD36 in macrophages. Results： The model group showed lower level of HDL-C than the blank control group, as well as higher level of TC, TG and LDL-C than the blank control group; larger atheromatous plaque was formed in aortic lumen, and a large number of lipid deposition was formed on aortic wall in model group, the model group showed lower macrophage ABCA1 mRNA and protein expression of peritoneal macrophages than the blank control group; ABCG1 expression did not change signifi- cantly in model group, and the model group showed higher CD36 mRNA and protein expression than the blank control group. Compared to the model group, the blood fat was ameliorated and the atherosclerotic plaque area in aortic lumen and lipid deposition on pipe wall were significantly reduced in the Tanshinone ⅡA group. The Tanshinone ⅡA group showed higher level of gene and protein expression of macrophage ABCA1, as well as lower level of CD36 gene and protein expression than the model group; The changes of ABCGI expression between the two groups showed no statistical difference. Conclusion： Tanshinone lI A can inhibit the formation of aortic plaque in ApoE-/- mice, and its mechanism may be related to the regulation of the expression of lipid and macrophage cholesterol efflux related gene ABCA1 and CD36.