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目的观察青心酮对小鼠动脉粥样硬化(AS)斑块中平滑肌细胞、内皮细胞、巨噬细胞Toll样受体4表达(Toll-like receptor 4,TLR4)的影响,及对载脂蛋白E基因敲除小鼠主动脉粥样硬化病变的影响。方法取小鼠胸主动脉,进行平滑肌细胞、内皮细胞的原代培养,RAW264.7(小鼠巨噬细胞)细胞株传代培养。实验分4组:空白对照组,脂多糖(LPS)组,青心酮组,辛伐他汀组。收集各组细胞总mRNA及蛋白,应用RT-PCR及Western-blot方法检测TLR4 mRNA及蛋白。取30只8周龄雄性ApoE(-/-)小鼠,随机分成3组:模型组(n=10),每天生理盐水灌胃;青心酮治疗组(n=10)经胃管灌注青心酮10mg·kg-1·d-1,辛伐他汀治疗组(n=10)经胃管灌注辛伐他汀10mg·kg-1·d-1。所有实验小鼠均饲以“西方类型膳食”饲料12周。剪取主动脉根部切片及HE染色观察主动脉粥样硬化病变情况。结果1×10-7mol·L-1青心酮组血管平滑肌细胞、内皮细胞、巨噬细胞TLR4 mRNA和蛋白含量明显低于未处理组(P<0.01);与辛伐他汀组比较无显著性差异;青心酮组AS病灶形成减少。结论青心酮可下调LPS活化的小鼠平滑肌细胞、内皮细胞、巨噬细胞TLR4mRNA和蛋白的表达,在一定程度上能减少AS病灶的形成,可能通过TLR4途径发挥其作用。
Objective To observe the effects of mependone on the expression of Toll-like receptor 4 (TLR4) in smooth muscle cells, endothelial cells and macrophages in atherosclerotic plaques of mice and the effect of adriamycin E gene knockout mice aortic atherosclerosis lesions. Methods Mouse thoracic aorta was taken for primary culture of smooth muscle cells and endothelial cells. The passage of RAW264.7 (mouse macrophage) cell line was subcultured. The experiment was divided into 4 groups: blank control group, lipopolysaccharide (LPS) group, Qingxin ketone group, simvastatin group. The total mRNA and protein of cells in each group were collected. The mRNA and protein of TLR4 were detected by RT-PCR and Western-blot. Thirty male ApoE (- / -) mice of 8 weeks old were randomly divided into 3 groups: model group (n = 10), normal saline intragastric administration; Simvastatin 10 mg · kg-1 · d-1 and simvastatin 10 mg · kg-1 · d-1 were intragastrically administered with simvastatin. All experimental mice were fed “western diet” for 12 weeks. Aortic root sections were cut and HE staining was used to observe the changes of aortic atherosclerosis. Results The mRNA and protein expressions of TLR4 in vascular smooth muscle cells, endothelial cells and macrophages in 1 × 10-7mol·L-1 macrophages were significantly lower than those in untreated group (P <0.01), but not in simvastatin group Differences; Qingxin ketone group AS lesion formation reduced. Conclusions Qingxin ketone can down-regulate the expression of TLR4mRNA and protein in LPS-activated mouse smooth muscle cells, endothelial cells and macrophages to a certain extent, reduce the formation of AS lesions and may play its role through the TLR4 pathway.