益肝降脂方预处理对酒精性脂肪肝SD大鼠诱导型一氧化氮合酶表达影响

Effects of Yigan Jiangzhi Formula in Pretreatment on Expression of Inducible Nitric Oxide Synthase of Alcoholic Fatty Liver SD Rats

目的:观察分析益肝降脂方(YGJZF)预处理对酒精性脂肪肝(AFLD)SD大鼠内源性保护物质诱导型一氧化氮合酶(iNOS)mRNA和蛋白表达的影响,探讨YGJZF预处理模拟或加强缺血预适应效应对肝脏保护作用的机制。方法:将116只SD大鼠随机分为模型组22只,中药干预组94只,模型组连续12周予高脂+酒精灌胃进行酒精性脂肪肝造模,中药组在模型组基础上用中药干预。分别于实验不同时段宰杀取样,检测大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、甘油三酯(TG)、总胆固醇(T-CHO)含量;实时荧光定量(PCR)和免疫印迹法(Western blot)分别检测大鼠肝脏iNOS mRNA和蛋白的表达;同时HE染色观察大鼠肝脏组织病理学变化。结果:模型组大鼠ALT、AST、TG、TC水平明显升高,与YGJZF用药3周组、YGJZF用药6周组、YGJZF用药9周组比较,差异有显著性意义(P<0.05);RT-PCR和Western blot显示模型组肝组织iNOS mRNA和蛋白呈现高表达,随着用药时间的延长,iNOSmRNA和蛋白表达逐渐下降;肝组织HE染色酒精模型组肝脏损伤严重,随着YGJZF用药时间的延长,肝脏损伤逐渐减轻。结论:YGJZF对酒精性脂肪肝保护作用机制可能是触发了肝脏的内源性保护传导路径的关键物质诱导型一氧化氮合酶(iNOS)的表达,模拟并加强了肝脏的缺血预适应效应。

Objective：To observe and analyze the impact of Yigan Jiangzhi Formula（YGJZF） in the pretreatment on the expression of endogenous protective substances inducible nitric oxide synthase（i NOS）m RNA and protein of alcoholic fatty liver SD rats and to explore YGJZF pretreatment simulation or strengthening the effect of ischemic preconditioning on liver protection mechanism. Method：116 SD rats were randomly divided into model group 22 and the Chinese intervention group 94. The model group was given fed high-fat diet and gastric perfusion of alcohol for 12 weeks to make AFL model and the Chinese intervention group added traditional Chinese medicine intervention on the basis of the model group. Completing the appropriate sampling slaughter at different times,we determined the contents of serum alanineaminotransferase（ALT）,aspartate minotransferase（AST）,triglyceride（TG）,total cholesterol（T-CHO）in rats. Real-time quantitative（PCR）and Western blot were used to detect the expressions of i NOS m RNA and protein in rat liver and HE staining was used to observe rat liver tissue＇s patnological changes. Result：The levels of ALT,AST,TG and TC increased significantly in model group and compared with YGJZF medication three weeks,six weeks and nine weeks groups,the difference was statistically significant（P〈0.05）. RT-PCR and Western blot displayed model group＇s liver tissue i NOS m RNA and protein expression gradually declined. HE staining of liver showed severe liver injury in model group with the extension of YGJZF medication time,the liver damage gradually reduced. Conclusion：The protection mechanism of YGJZF on alcoholic fatty liver may be triggering the endogenous protection conduction path of key material i NOS as well as simulating and strengthening the liver ischemic preconditioning effect.