化瘀祛痰方对ApoE^(-/-)AS小鼠主动脉G6PD/NF-κB表达及炎症水平的影响

Effect of Huayu Qutan Formula(化瘀祛痰方)on G6PD/NF-κB Pathway in ApoE^(-/-)AS Mice Aortic and Inflammation

目的基于G6PD/NF-кB表达,探讨化瘀祛痰方对ApoE^(-/-)动脉粥样硬化(athrosclerotic,AS)小鼠斑块及炎症反应的影响及机制研究。方法24只ApoE基因敲除(ApoE^(-/-))小鼠随机分为模型组、化瘀祛痰方组、辛伐他汀组,给予高脂饲料喂养12周。选用8只C57BL/6J小鼠作为空白组,给予普通饲料喂养。化瘀祛痰方组以及辛伐他汀组分别给予相应的药物灌胃,空白组与模型组给予等量的生理盐水。4周后采用酶联免疫法(ELISA)检测血清甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)的含量;采用苏木素-伊红(HE)染色观察主动脉斑块形态学改变;ELISA法检测各组小鼠血清中IL-6、IL-1β、TNF-α水平;Western Blot法及实时荧光定量Q-PCR法检测主动脉IL-6、IL-1β、TNF-α、G6PD、NF-κB蛋白及mRNA的表达。结果与空白组比较,模型组小鼠血清TC、TG和LDL-C含量显著升高(P<0.05或P<0.01),HDL-C含量无明显改变(P>0.05),主动脉内膜增厚,管腔狭窄,大量粥样斑块形成,血清中IL-6、IL-1β、TNF-ɑ水平显著升高(P<0.05或P<0.01),主动脉内IL-6、IL-1β、TNF-ɑ、G6PD、NF-κB蛋白及mRNA水平显著升高(P<0.05或P<0.01);与模型组相比,化瘀祛痰组及辛伐他汀组小鼠血清TC、TG和LDL-C含量显著降低(P<0.05或P<0.01),HDL-C含量无明显改变(P>0.05),主动脉斑块较模型组明显减小,血清中IL-6、IL-1β、TNF-ɑ水平显著降低(P<0.05或P<0.01),主动脉内IL-6、IL-1β、TNF-ɑ、G6PD、NF-κB蛋白及mRNA水平显著降低(P<0.05或P<0.01)。结论化瘀祛痰方能够减小主动脉斑块大小,延缓AS进展,其机制可能与其调控G6PD/NF-κB表达,降低G6PD、NF-κB蛋白水平、抑制炎症反应有关。

Objective Based on G6PD/NF-κB pathway,to explore the effect and mechanism of Huayu Qutan Formula(化瘀祛痰方)on plaque and inflammation in ApoE^(-/-)atherosclerotic mice.Methods Twenty-four ApoE gene knockout mice were randomly divided into three groups:model group,Huayu Qutan Formula group and simvastatin group.The mice were fed with high-fat diet for 12 weeks.Eight C57BL/6J mice were selected as blank group and fed with common diet.The corresponding drugs were given in the Huayu Qutan Formula group and simvastatin group by gavage,and the same amount of normal saline was given to the blank group and the model group.Four weeks later,the contents of serum triglyceride(TG),total cholesterol(TC),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol(LDL-C)were detected by ELISA method.The morphological changes of aortic plaque were observed by hematoxylin-eosin(HE)staining,and the levels of IL-6,IL-1βand TNF-αin serum of mice in each group were detected by ELISA method.The levels of IL-6,IL-1β,TNF-α,G6PD,NF-κB protein and mRNA in aorta were detected by Western blot and Real-time Q-PCR.Results Compared with those of the blank group,the contents of serum TC,TG and LDL-C in the model group were increased significantly(P<0.05,P<0.01),but the content of HDL-C had no significant change(P>0.05).The intima of aorta thickened,the lumen narrowed and a large number of atherosclerotic plaques formed,and the levels of serum IL-6,IL-1β,TNF-α.IL-6,IL-1β,TNF-α,G6PD,NF-κB and mRNA in aorta were significantly increased(P<0.05,P<0.01).Compared with those of the model group,the contents of serum TC,TG and LDL-C in the Huayu Qutan Formula group and simvastatin group were decreased significantly,but the content of HDL-C did not change significantly(P>0.05),and the aortic plaque in the Huayu Qutan Formula group and simvastatin group was significantly lower than that in the model group(P<0.05,P<0.01).The levels of IL-6,IL-1βand TNF-αin serum and the levels of IL-6,IL-1β,TNF-α,G6PD,NF-κB protein and mRNA in aorta were significantly decreased(P<0.05,P<0.01).Conclusion Huayu Qutan Formula can reduce the size of aortic plaque and delay the progression of AS.Its mechanism may be related to its regulation of G6PD/NF-κB pathway,reduction of G6PD and NF-κB protein levels and inhibition of inflammatory reaction.