补阳还五汤加减抑制ApoE^(-/-)小鼠动脉粥样硬化病变形成及降低血清炎症因子水平的机制

Study on the mechanism of Buyang Huanwu decoction in inhibiting atherosclerotic lesion formation and reducing serum inflammatory factors in ApoE^(-/-)mice

[目的]基于Wnt/β连环蛋白(β-catenin)信号通路探讨补阳还五汤加减通过调控血管平滑肌细胞的表型转化及血清炎症因子水平,发挥抗动脉粥样硬化(As)的作用。[方法]将50只ApoE^(-/-)小鼠随机分为模型组、补阳还五汤加减(低、中、高剂量)组及阿托伐他汀组,每组10只;10只C57BL/6J小鼠设为对照组,给予普通饲料喂养,其余5组小鼠给予高脂饲料喂养8周进行As模型复制,第9周开始灌胃,连续灌胃4周,12周后分离小鼠主动脉根部,制备石蜡切片,油红O染色观察主动脉窦脂质含量;免疫组织化学法检测骨桥蛋白(OPN)表达水平;ELISA法检测小鼠血清中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、单核细胞趋化蛋白1(MCP-1)的水平;Western blot检测小鼠主动脉中Wnt1、β-catenin及c-myc蛋白的表达;RT-PCR法检测小鼠主动脉中Wnt1、β-catenin基因的表达。[结果]补阳还五汤加减(低、中、高剂量)能够降低As模型小鼠主动脉窦脂质含量及OPN表达;降低小鼠血清IL-6、TNF-α、IL-1β、MCP-1水平;并可下调小鼠主动脉中Wnt1、β-catenin蛋白及基因的表达。[结论]补阳还五汤加减通过调控炎症反应发挥抗As的作用,部分机制可能与抑制Wnt/β-catenin信号通路活化有关。

Aim To explore the effect of Buyang Huanwu decoction on anti-atherosclerosis by regulating the phenotypic transformation of vascular smooth muscle cells and serum inflammatory response based on Wnt/β-catenin signaling pathway.Methods 50 ApoE^(-/-)mice were randomly divided into model group,Buyang Huanwu decoction modified(low,medium and high dose)groups,and atorvastatin group,with 10 mice in each group;10 C57BL/6J mice with the same genetic background were set as the control group.The mice of control group were fed with regular forages,while the mice of other five groups were fed with high-fat forages for 8 weeks for model replication.From the 9th week,they were continuously gavaged for 4 weeks.After 12 weeks,the mouse aortic roots were isolated and paraffin sections were prepared.Oil red O staining was used to observe the lipid content of aortic sinus.Immunohistochemical method was used to detect the expression level of bone bridge protein(OPN).Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and monocyte chemoattractant protein-1(MCP-1)in serum.Western blot was used to detect the protein expressions of Wnt1,β-catenin and c-myc in mice aorta.RT-PCR was used to detect the expressions of Wnt1 andβ-catenin gene.Results Buyang Huanwu decoction modified(low,medium and high dose)could reduce the lipid content and OPN expression in the aortic sinus of As model mice,reduce serum levels of IL-6 and TNF-α,IL-1βand MCP-1 in mice,and downregulate the expression of Wnt1 andβ-catenin protein and gene.Conclusion Buyang Huanwu decoction modified exerts an anti-atherosclerosis effect by regulating inflammatory response,and some of the mechanisms may be related to inhibiting the activation of the Wnt/β-catenin signaling pathway.