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目的:探究加味补阳还五汤对于载脂蛋白E基因敲除小鼠的Toll样受体4(TLR4)及髓样分化因子88(My D88),肿瘤坏死因子受体相关因子-6(TRAF-6),核转录因子-κB(NF-κB)表达的影响,讨论加味补阳还五汤对于动脉粥样硬化的防治机制。方法:将30只雄性载脂蛋白E基因敲除小鼠随机分为模型组,阿托伐他汀组,加味补阳还五汤组,每组10只,以10只C57BL/6J小鼠设空白组。除空白组正常饮食饮水外,每组给予高脂饲料喂养8周。空白组及模型组每日给与0.9%Na Cl溶液灌胃,阿托伐他汀组及加为补阳还五汤组每日以相应药物灌胃。第9周后采用生化检测方法检测血清总胆固醇(TC),甘油三酯(TG),高密度脂蛋白(HDL-C)和低密度脂蛋白(LDL-C)。采用苏木素-伊红(HE)染色方法观察,并测量斑块所占管腔面积之比。以实时荧光定量PCR(Real-time PCR)检测TLR4及My D88,TRAF-6,NF-κB的mRNA表达;蛋白质印迹(Western blot)法测定其蛋白表达的变化。结果:血脂水平方面,与模型组比较加味补阳还五汤组与阿托伐他汀组均能降低载脂蛋白E基因敲除小鼠TC,TG,HDL-C,LDL-C水平,同时升高HDL-C水平(P<0.01);镜下观察加味补阳还五汤组与阿托伐他汀组均能较模型组斑块有所减少,主动脉血管细胞排列较整齐,炎性细胞浸润减轻,小鼠的主动脉斑块在血管的占比均有不同程度地降低(P<0.01)。mRNA与蛋白水平上与模型组比较加味补阳还五汤与阿托伐他汀组均能有效降低TLR4及My D88,TRAF-6,NF-κB mRNA与蛋白的表达(P<0.05,P<0.01)。结论:加味补阳还五汤对于AS的发生发展有预防作用,其机制可能与抑制TLR4及其下游信号转导通路主要元件有关。
Objective: To investigate the effects of Modified Buyang Huanwu Decoction (TLR4) on TLR4, My D88, TRAF - 6 (TRAF) in apolipoprotein E knockout mice, -6), and nuclear factor kappa B (NF-κB) expression, to discuss the mechanism of Jiaweibuhuanyuanwu decoction on atherosclerosis. Methods: Thirty male Apo E gene knockout mice were randomly divided into model group, atorvastatin group and Jiawei Buyang Huanwu Decoction group, with 10 mice in each group. 10 C57BL / 6J mice were blank group. In addition to the normal diet of the blank group of drinking water, each group fed high-fat diet for 8 weeks. The rats in the blank group and model group were orally gavaged with 0.9% NaCl solution daily. Atorvastatin group and BYHWD group were given the corresponding drugs daily. After 9 weeks, serum total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL-C) and low density lipoprotein (LDL-C) were detected by biochemical methods. The hematoxylin-eosin (HE) staining method was used to observe the area of plaque in the lumen area. The mRNA expressions of TLR4, My D88, TRAF-6 and NF-κB were detected by Real-time PCR. The protein expression of TLR4 and My-D88 were detected by Western blot. Results: Compared with the model group, the levels of TC, TG, HDL-C and LDL-C in the BYDH group and atorvastatin group were significantly lower than those in the model group High HDL-C levels (P <0.01). The microscopic observation showed that the levels of plaque in Jiawe BuYuanHuaWu Decoction group and atorvastatin group were lower than those in model group, the aortic vascular cells were arranged neatly, inflammatory cell infiltration Reduce the proportion of aortic plaque in blood vessels of mice decreased to some extent (P <0.01). (P <0.05, P <0.01). Compared with the model group, both JiaYuHuWuWu Decoction and Atorvastatin group can effectively reduce the mRNA and protein expressions of TLR4, My D88, TRAF-6 and NF- ). Conclusion: Jiaweibu Bu Yang Huanwu Decoction can prevent the occurrence and development of AS, which may be related to the inhibition of TLR4 and its downstream signal transduction pathway.