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目的探讨葱白提取物治疗高脂饮食诱导的非酒精性肝病大鼠可能的分子机制。方法采用高脂饮食建立大鼠非酒精性脂肪肝模型,并应用小干扰RNA技术抑制转染组大鼠辅激活因子(peroxisome proliferator-activated receptor-γcoactivator`1α,PGC-1α)的表达,将48只Wistar大鼠随机分为6组,观察各组大鼠的一般情况,计算肝指数,油红染色法观察各组大鼠肝脏组织的病理形态学变化,并运用RT-PCR法和Western-blot法分别检测PGC-1α和脂肪酸β氧化限速酶肉毒碱棕榈酰基转移酶-1(carnitine palmitoyltransferase-1,CPT-1)的表达水平。结果各组大鼠体重无明显差异,模型组精神较差,偶见腹泻便溏情况,活动度明显不及葱白治疗组。与正常对照组相比,模型组大鼠肝指数明显升高(P<0.05);与模型组比较,葱白组肝指数下降(P<0.05),转染葱白组与模型组肝指数差异不明显。从油红染色可见模型组大鼠肝细胞内较多橘红色脂滴聚集,细胞肿胀明细;葱白组胞内脂滴明显减少;转染葱白组胞内脂滴数量介于模型组和葱白治疗组之间。PGC-1α和CPT-1的表达情况:与空转组比,转染组PGC-1α的表达水平降低(P<0.05),CPT-1的表达水平也下降(P<0.05)。和正常组比,模型组PGC-1α与CPT-1的表达水平均降低(P<0.05);和模型组比,葱白组PGC-1α与CPT-1的表达水平均增加(P<0.05),转染葱白组两者的表达则与模型组无明显差异(P>0.05)。结论辛温通阳中药葱白提取物治疗NAFLD依赖PGC-1α的转录活化水平,从而调控肝细胞脂肪酸β氧化发挥作用。
Objective To investigate the possible molecular mechanism of scallion extract in the treatment of non-alcoholic liver disease induced by high-fat diet in rats. Methods A rat model of non-alcoholic fatty liver was established by high-fat diet. The expression of peroxisome proliferator-activated receptor-γcoactivator (PAH) 1α and PGC-1α in transfected rats was inhibited by small interfering RNA Only Wistar rats were randomly divided into 6 groups. The general conditions of rats in each group were observed. The liver index was calculated. The histopathological changes of liver tissues were observed by oil red staining. RT-PCR and Western-blot Method was used to detect the expression level of PGC-1α and carnitine palmitoyltransferase-1 (CPT-1), a fatty acid β-oxidation rate-limiting enzyme. Results There was no significant difference in body weight of rats in each group. The spirit of the model group was poor, occasional diarrhea stool situation, activity was significantly less than the light blue treatment group. Compared with the normal control group, the liver index of the model group was significantly increased (P <0.05). Compared with the model group, the liver index decreased (P <0.05), and the liver index of the model group was not significantly different . From the oil red staining, more lipid droplets were accumulated in the hepatocytes of the model group, and the details of the cell swelling were obvious. The intracellular lipid droplets in the light group were significantly decreased. The intracellular lipid droplets transfered from the model group to the light blue treatment group between. The expression of PGC-1α and CPT-1 in the transfected group was lower than that in the untransfected group (P <0.05), and the expression of PGT-1 was also decreased (P <0.05). Compared with the normal group, the expression levels of PGC-1α and CPT-1 in the model group were significantly lower than those in the model group (P <0.05) There was no significant difference between the two groups (P> 0.05). Conclusion Xinwen Tongyang extract of Chinese white extract treatment of NAFLD dependent on PGC-1α transcriptional activation level, and thus regulate fatty liver β fatty acid oxidation play a role.