脾虚痰浊证巴马小型猪心肌组织LKB1/MPK信号通路的变化

Change of LKB1/AMPK Pathway in Cardiac Muscle of Bama Minipigs with Spleen Deficiency and Phlegm Turbidity Syndrome

目的:探讨脾虚痰浊证巴马小型猪心肌物质代谢相关通路LKB1/AMPK的变化。方法:普通级雄性巴马小型猪10只随机分为正常组、脾虚痰浊组,每组5头。采用疲劳过度联合高脂单笼饲养建立小型猪脾虚痰浊证模型。采用实时荧光定量PCR法检测心肌组织腺苷单磷酸活化蛋白激酶(AMPK)、磷酸化腺苷单磷酸活化蛋白激酶(p AMPK)、肝脏激酶B1(LKB1)mRNA表达,应用Western blotting法检测心肌组织AMPK、p AMPK、LKB1蛋白表达水平。结果:与正常组小型猪相比,脾虚痰浊组小型猪心肌组织AMPK、p AMPK mRNA表达水平显著下降(P<0.05),AMPK、p AMPK蛋白表达水平亦明显下降(P<0.01);与正常组小型猪相比,脾虚痰浊组小型猪心肌组织LKB1mRNA及蛋白表达水平显著下降(P<0.01)。结论:心肌组织LKB1/AMPK信号通路相关分子表达下调可能为脾虚痰浊证的分子生物学基础。

Objective： To investigate the change of LKB1/AMPK pathway in cardiac muscle of Bama Minipigs with spleen deficiency and phlegm turbidity syndrome. Methods： Ten male common grade Bama minipigs were randomly divided into normal group, spleen deficiency and phlegm turbidity group. The model of spleen deficiency and phlegm turbidity syndrome was established by the combination of fatigue and high - fat feeding separately. The expressions of AMPK, pAMPK and LKB1 mRNA were detected by real - time fluorescence quantitative PCR method and the expression levels of AMPK, pAMPK and LKB1 protein in cardiac muscle tissue were detected by Western blotting method. Results： Compared with the normal group, in the group of spleen deficiency and phlegm tuibidity, the expression levels of AMPK and pAMPK mRNA in the myocardium were significantly decreased （P 〈 0.05 ）. The expression levels of AMPK and pAMPK protein were decreased （P 〈 0. 01 ）. The expression levels of LKBlmRNA and protein in the myocardium were significantly decreased （P 〈 0. 01 ）. Conclusion ： The down regulated expression of AMPK/LKB1 signaling pathway related molecules in the myocardium of Bama Minipigs with spleen deficiency and phlegm turbidity syndrome may be the molecular biological basis of spleen deficiency and phlegm turbidity syndrome.