钙依赖性酪氨酸激酶2和微囊蛋白1传导路在酒精性心肌病中的作用与药物干预研究

目的 探讨钙依赖性酪氨酸激酶2(PYK2)和微囊蛋白1传导路在酒精性心肌病(ACM)中的作用及与氧化应激和心肌纤维化的关系.方法 健康成年杂种犬28只,分为乙醇组、缬沙坦+乙醇组、肉毒碱+乙醇组和正常对照组,每组7只.检测各组实验动物心功能.比色法测定心肌组织超氧化物歧化酶(SOD)、谷胱甘肽过氧物酶(GSH-Px)和丙二醛(MDA)含量.ELISA法测定心肌组织Ⅰ型、Ⅲ型胶原水平.免疫组化法观察PYK2和微囊蛋白表达.结果 SOD、GSH-Px与对照组(56.6±17.0) U/mg蛋白、(488±33) U/g蛋白]比较,乙醇组、缬沙坦+乙醇组和肉毒碱+乙醇组明显下降SOD (U/mg蛋白)分别为28±5、33±13、33±10;GSH-Px(U/g蛋白)分别为188±37、362±29、282±29];MDA比对照组(6.4±0.7) nmol/mg蛋白]乙醇组和肉毒碱+乙醇组升高分别为(19.4±3.0) nmol/mg蛋白、(10.8±2.1) nmol/mg蛋白].与对照组比较,Ⅰ型胶原含量在乙醇组、缬沙坦+乙醇组和肉毒碱+乙醇组心肌组织中增加;Ⅲ型胶原含量在乙醇组和肉毒碱+乙醇组心肌组织中增加(均P<0.01).乙醇组、缬沙坦+乙醇组和肉毒碱+乙醇组心肌组织PYK2蛋白表达增加、微囊蛋白表达减少,与对照组比较差异有统计学意义(均P<0.01).结论 PYK2和微囊蛋白1传导路可能通过调节氧化应激和胶原合成而发挥心肌纤维化调控作用.

Abstract：

Objective To explore the protein expressions of PYK2 (protein-rich tyrosine kinase-2) and caveolin-1 in alcoholic cardiomyopathy (ACM) dog model and the effect of early drug intervention.Methods A total of 28 adult dogs were randomly divided into 4 groups: (i): alcohol-fed;(ii): alcohol plus valsartan, an angiotensin receptor blocker (ARB);(iii): alcohol plus carnitine;(iv): control group. At Month 6, the cardiac functions of all animals were evaluated by echocardiography. The concentrations of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in cardiac tissue were detected by chromometry. The protein expressions of collagen typesⅠand Ⅲ were determined by ELISA (enzyme-linked immunosorbent assay) while those of PYK2 and caveolin-1 evaluated by immunohistochemistry. The internal relationship of PYK2, caveolin-1, MDA and collagen typesⅠ& Ⅲ was analyzed.Results Compared with the control group, the contents of SOD and GSH-Px significantly decreased in the alcohol, valsartan and carnitine groups while MDA significantly increased in the alcohol and carnitine groups. The relative contents in each group were as follows: MDAalcohol: (28±5)U/mg Pro;valsartan: (33±13)U/mg Pro;carnitine:(33±10)U/mg Pro], GSH-Pxalcohol:(188±37)U/g Pro;valsartan:(362±29)U/g Pro;carnitine:(282±29) U/g Pro and MDA alcohol:(19.4±3.0)nmol/mg Pro;carnitine:(10.8±2.1)nmol/mg Pro].Compared with the control group, the concentration of collagen typeⅠsignificantly increased in the alcohol, valsartan and carnitine groups. In the alcohol-fed group, theprotein expression of PYK2 gradually increased with the progression of ACM while that of caveolin-1 stayed at a low level. Through the interventions of valsartan and carnitine, the protein expression of PYK2 significantly increased while that of caveolin-1 significantly decreased (all P<0.01). Conclusion Alcohol promotes the myocardial oxidative stress and fibrosis through an elevated expression of PYK2 and a lowered expression of caveolin-1. Then a deterioration of cardiac structures and functions occur

Roles of PYK2 and caveolin-1 in alcoholic cardiomyopathy and the effect of drug intervention

Objective To explore the protein expressions of PYK2 (protein-rich tyrosine kinase-2) and caveolin-1 in alcoholic cardiomyopathy (ACM) dog model and the effect of early drug intervention.Methods A total of 28 adult dogs were randomly divided into 4 groups: (i): alcohol-fed;(ii): alcohol plus valsartan, an angiotensin receptor blocker (ARB);(iii): alcohol plus carnitine;(iv): control group. At Month 6, the cardiac functions of all animals were evaluated by echocardiography. The concentrations of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in cardiac tissue were detected by chromometry. The protein expressions of collagen typesⅠand Ⅲ were determined by ELISA (enzyme-linked immunosorbent assay) while those of PYK2 and caveolin-1 evaluated by immunohistochemistry. The internal relationship of PYK2, caveolin-1, MDA and collagen typesⅠ& Ⅲ was analyzed.Results Compared with the control group, the contents of SOD and GSH-Px significantly decreased in the alcohol, valsartan and carnitine groups while MDA significantly increased in the alcohol and carnitine groups. The relative contents in each group were as follows: MDAalcohol: (28±5)U/mg Pro;valsartan: (33±13)U/mg Pro;carnitine:(33±10)U/mg Pro], GSH-Pxalcohol:(188±37)U/g Pro;valsartan:(362±29)U/g Pro;carnitine:(282±29) U/g Pro and MDA alcohol:(19.4±3.0)nmol/mg Pro;carnitine:(10.8±2.1)nmol/mg Pro].Compared with the control group, the concentration of collagen typeⅠsignificantly increased in the alcohol, valsartan and carnitine groups. In the alcohol-fed group, theprotein expression of PYK2 gradually increased with the progression of ACM while that of caveolin-1 stayed at a low level. Through the interventions of valsartan and carnitine, the protein expression of PYK2 significantly increased while that of caveolin-1 significantly decreased (all P<0.01). Conclusion Alcohol promotes the myocardial oxidative stress and fibrosis through an elevated expression of PYK2 and a lowered expression of caveolin-1. Then a deterioration of cardiac structures and functions occur