山茱萸对糖尿病心肌病大鼠的干预作用及机制

EFFECT AND ITS MECHANISM OF FRUCTUS CORNI ON DIABETIC CARDIOMYOPATHY IN RATS

目的探讨山茱萸对糖尿病心肌病大鼠的干预作用及机制。方法采用腹腔注射链脲佐菌素方法构建糖尿病大鼠模型。将50只Sprague-Dawley大鼠随机分为正常对照组（N组）、糖尿病组（D组）、山茱萸低剂量组（FC-L组,400mg/kg）、山茱萸高剂量组（FC-H组,800mg/kg）及氟伐他汀组（Flu组）,每组10只。12周后测体质量并计算心体比（H/B）和左心室质量指数（LVMI）。应用流式细胞仪检测心肌组织中活性氧（ROS）的表达,生化法检测大鼠血清中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-px）活性和心肌细胞中谷胱甘肽（GSH）含量,Western blot及real time PCR检测心肌组织中p-Akt蛋白及P22phox、p47phox、FN、ColⅢ和核因子-κB（NF-κB）mRNA表达。结果与对照组相比,模型组H/B、LVMI、ROS及P22phox、p47phox、FN、ColⅢ、NF-κB和p-Akt表达均显著升高（F=295.274～543.871,P〈0.05）,血清SOD、GSH-px和心肌细胞中的GSH含量均显著降低（F=325.274～415.287,P〈0.05）。山茱萸及氟伐他汀干预后,Flu组、FC-L组和FC-H组大鼠的心肌H/B、LVMI、ROS、P22phox、p47phox、FN、ColⅢ、NF-κB和p-Akt表达均显著低于模型组（F=112.557～478.684,P〈0.05）,血清SOD、GSH-px和心肌细胞中GSH含量显著升高（F=198.547～347.574,P〈0.05）,且均呈剂量依赖性。结论山茱萸干预可以减轻糖尿病大鼠心肌病变,可能与减少心肌组织氧化应激水平,下调NADPH氧化酶P22phox和p47Phox亚基与p-Akt表达有关。

Objective To explore the effect and mechanism of Fructus Corni on diabetic cardiomyopathy in rats. Methods Employing intraperitoneal injection of Streptozotocin to create a model of diabetes in rats,50 Sprague-Dawley rats were then equally randomized to five groups as：normal group（N group）,diabetes mellitus group（D group）,Fructus Corni low dose group（FC-L group,400mg/（kg·d））,and Fructus Corni high dose group（FC-H group,800mg/（kg·d））,and fluvastatin-positive control group（Flu group）.The weight of rats was measured 12 weeks later,the heart to body weight ratio（H/B）and the left ventricle mass index（LVMI）were calculated.Employing flow cytometer to detect the ROS expression in cardiac muscle,biochemical method to detect the levels of serum SOD,GSH-px and GSH in cardiac muscle cells,and Western blotting and real time PCR to detect the expressions of p-Akt protein in cardiac muscle tissue and the expressions of P22 phox,p47phox,NF-κB and ColⅢ.Results Compared with the normal control group,the H/B,LVMI,and expressions of ROS,P22 phox,p47phox,FN,ColⅢ,NF-κB and p-Akt in the N group elevated（F=295.274-543.871,P〈0.05）,and serum SOD,GSH-px and GSH in cardiac muscle cells declined（F=325.274-415.287,P〈0.05）.After treatment with Fructus Corni,H/B and LVMI,the expressions of ROS,P22 phox,p47phox,FN,ColⅢ,NF-κB and p-Akt in Flu group,FC-L group and FC-H group were lower versus the N group（F=112.557-478.684,P〈0.05）,and serum levels of SOD,GSH-px,and GSH in cardiac muscle cells elevated（F=198.547-347.574,P〈0.05）,showing dose dependent in manner. Conclusion Fructus Corni can relieve the myocardial lesion of diabetic rats,the mechanism may be associated with decreasing myocardial oxidative stress and down-regulating the expressions of NADPH oxidase p22 phox and p47 phox subunits.