黄连温胆汤调控PI3K/Akt/mTOR信号通路干预ApoE-/-小鼠动脉粥样硬化的作用机制

目的观察黄连温胆汤对载脂蛋白E(ApoE)^-/-小鼠动脉粥样硬化的干预效应,从磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物西罗莫司靶蛋白(mTOR)信号通路探讨其机制。方法采用高脂饲料饲养ApoE^-/-小鼠4周构建动脉粥样硬化模型。采用全自动生化分析仪检测小鼠血脂水平;采用苏木精-伊红(HE)染色、油红O染色、Masson染色观察主动脉斑块情况;采用免疫组化法检测小鼠主动脉根部PI3K、Akt、mTOR蛋白表达水平。结果与模型组比较,黄连温胆汤高剂量组总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)均降低(P<0.05)。HE染色显示:模型组主动脉根部斑块明显,呈易损斑块特征,各给药组斑块有不同程度减轻;油红O和Masson染色显示:与模型组比较,雷帕霉素组、黄连温胆汤高剂量组主动脉根部斑块面积减小,胶原纤维含量升高(P<0.05),易损斑块趋于稳定。蛋白免疫组化结果显示:黄连温胆汤中剂量组、黄连温胆汤高剂量组小鼠主动脉PI3K、Akt、mTOR蛋白表达水平降低(P<0.05)。结论黄连温胆汤具有降脂和抗动脉粥样硬化的作用,其机制可能与抑制PI3K/Akt/mTOR信号通路,诱导巨噬细胞自噬有关。

Mechanism of Huanglian Wendan Decoction Regulating PI3K/Akt/mTOR Signal Pathway to Intervene Atherosclerosis of ApoE-/- Mice

Objective To observe the intervention effect of Huanglian Wendan Decoction on atherosclerosis in apolipoprotein E(ApoE)^-/- mice,and to explore the mechanism of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target protein(mTOR)signaling pathway.Methods ApoE^-/- mice were fed with high fat diet for 4 weeks to model atherosclerosis.The blood lipid of mice were detected by automatic biochemical analyzer.The status of aortic plaque was observed by hematoxylin-eosin(HE)staining,oil red O staining,and Masson staining.The proteins expressions of PI3K,Akt,and mTOR in aorta root of mice were detected by immunohistochemistry.Results Compared with the model group,the levels of total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in Huanglian Wendan Decoction high-dose group were decreased significantly(P<0.05).HE staining showed that aortic root plaque in the model group was obvious,showing vulnerable plaque characteristics,and the plaques were alleviated degrees in each administration group.Oil red O and Masson staining showed that,compared with the model group,the area of aortic root plaque in the rapamycin group and Huanglian Wendan Decoction high-dose group was decreased,the content of collagen fiber was increased(P<0.05),and the vulnerable plaque tended to be stable.Immunohistochemical results showed that the protein expressions of PI3K,Akt,and mTOR in aorta of Huanglian Wendan Decoction middle-dose group and Huanglian Wendan Decoction high-dose group of Huanglian Wendan Decoction were decreased(P<0.05).Conclusion Huanglian Wendan Decoction possessed lipid-lowering and anti-atherosclerosis effects,and the mechanism might be related to the inhibition of PI3K/Akt/mTOR signaling pathway,inducing macrophage autophagy.