健脾消积法对非酒精性脂肪性肝病模型大鼠的抗氧化作用

Antioxidant Effect of Spleen-sdtrengthening and Accumulation-removing Therapy for Non-alcoholic Fatty Liver Disease in Rats

【目的】探讨健脾消积法对非酒精性脂肪性肝病(NAFLD)模型大鼠的抗氧化机制。【方法】将46只大鼠随机分为正常组(N=10)、模型组(N=8)、阳性对照组(N=9)、中药低剂量组(N=9)、中药高剂量组(N=10)等5组。给予大鼠喂饲高脂饲料诱导NAFLD大鼠模型。自造模第1天起,正常组、模型组大鼠给予等体积生理盐水灌胃,阳性对照组给予复方蛋氨酸胆碱溶液(剂量为0.38 g·kg^(-1)·d^(-1))灌胃,中药低、高剂量组给予加减丹溪枳术丸溶液(剂量分别为18、36 g·kg^(-1)·d^(-1))灌胃,共给药10周。采用常规油红O法观察肝组织形态学改变,采用实时荧光定量PCR检测肝组织核因子NF-E2相关因子(nuclear factor erythroid 2-related factor 2,Nrf2)、谷氨酰半胱氨酸合酶(γ-glutamylcysteine synthethase,γ-GCS)mRNA表达,采用硫代巴比妥酸法检测肝组织丙二醛(MDA)含量,黄嘌呤氧化酶法检测肝组织超氧化物歧化酶(SOD)含量。【结果】与模型组比较,中药高剂量组大鼠肝脏脂肪变性减轻,Nrf2 mRNA、γ-GCS mRNA表达水平及SOD含量显著升高,MDA含量显著降低(均P<0.05或P<0.01)。【结论】健脾消积法可有效干预NAFLD大鼠,其机制可能与增强Nrf2、γ-GCS表达,减轻氧化应激有关。

Objective To observe the antioxidant mechanism of spleen-strengthening and accumulation-removing therapy for treating non-alcoholic fatty liver disease（NAFLD）rats. Methods Forty-six rats were randomly dividedin to 5 groups,namely normal group（N = 10）,model group（N = 8）,positive control group（N = 9）,low-dose Chinese medicine group（N = 9）,high-dose Chinese medicine group（N = 10）. From the first modeling day,the positive control group was given intragastric administration of compound methionine choline solution（at the dosage of 0.38 g·kg^-1·d^-1）,the rats in the normal group and the model group were given intragastric administration of the same volume of normal saline, and the low-and high-dose Chinese medicine groups were given intragastric administration of 18,36 g ·kg^-1·d^-1 of modified Danxi Zhizhu Pills,respectively,the treatment covering 10 weeks. And then the conventional oil red O method was used to observe the morphological changes of liver tissues,real-time fluorescence quantitative PCR was used to detect the expression levels of nuclear factor erythroid 2-related factor 2（Nrf2）and glutamylcysteine synthase（γ-GCS）mRNA in liver tissues,thiobarbituric acid method was used to detect hepatic malondialdehyde（MDA）, and xanthine oxidase method was used to detect hepatic superoxide dismutase（SOD）. Results Compared with the model group, the hepatic steatosis was significantly relieved,the mRNA expression levels of Nrf2,γ-GCS and the content of SOD were significantly increased,and the content of MDA was significantly decreased in the high-dose Chinese medicine group（all P〈0.05 or P〈0.01）. Conclusion Spleen-strengthening and accumulation-removing therapy is effective for NAFLD rats,whose mechanism may be related to enhancing Nrf2,γ-GCS expression,and reducing oxidative stress.