活血祛湿方对纯化高脂饲料诱导大鼠非酒精性脂肪性肝炎模型的影响

Effects of Huoxue Qushi Formula on NASH Rats Model Induced by Purified High Fat Diet

目的:观察不同剂量活血祛湿方对纯化高脂饲料诱导的大鼠非酒精性脂肪性肝炎(NASH)模型的治疗作用,探讨活血祛湿方治疗NASH的作用机制。方法:纯化高脂饲料制备Wistar大鼠NASH模型8周,随机分为罗格列酮组(3 mg·kg-1·d-1),活血祛湿方组高、中、低剂量组(32,16,8 g·kg-1·d-1)及模型组,另设正常组。第9周开始ig给药;正常组及模型组均给予等量生理盐水ig。药物干预4周后行血清生化指标,釆用比色法检测血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),甘油三酯(TG),总胆固醇(TC)及高密度脂蛋白胆固醇(HDL-C)含量,放射免疫法检测血清空腹血糖(FBG),空腹胰岛素(FINS),酶联免疫吸附法(ELISA)法检测炎症因子肿瘤坏死因子-α(TNF-α),白细胞介素-10(IL-10)及IL-6;检测肝组织中超氧化物岐化酶(SOD)活性及丙二醛(MDA)含量;苏木素-伊红(HE)染色观察肝脏组织病理学变化;卵白素-生物素-酶复合物染色(ABC)法检测肝脏组织细胞色素P4502E1(CYP2E1)含量及分布;逆转录-聚合酶链式反应(RT-PCR)法检测过氧化物酶体增殖剂活化受体α(PPARα)mRNA表达。结果:与正常组比较,模型组大鼠血清ALT,AST,TG,TC水平及FBG,FINS,TNF-α及IL-6含量明显升高,HDL-C,IL-10含量明显降低,肝组织SOD活性明显降低,MDA及CYP2E1含量明显升高,肝脏PPARαmRNA表达明显降低,均具有明显的统计学差异(P<0.05);与模型组比较,活血祛湿方各剂量组均能够改善大鼠NASH模型各项血清生化指标,降低FBG,FINS水平,明显提高肝脏PPARαmRNA表达,明显增加肝脏中SOD活性及IL-10的含量,明显减少肝脏MDA,CYP2E1,TNF-α及IL-6含量,与模型组比较,差异有统计学意义(P<0.05)。病理结果显示,肝脏组织病理损伤明显改善。结论:活血祛湿方能够有效多途径治疗NASH,改善胰岛素抵抗,减少脂质过氧化反应,抑制炎症反应可能是其治疗NASH的机制。

Objective： To observe the effect of Huoxue Qushi formula on the non-alcoholic steatohepatitis（ NASH） model rat and explore it possible mechanism. Method： All Wistar rats of NASH model were induced by feeding high fat diet for 8 weeks,and they were randomly divided into the rogridone group（ 3mg·kg- 1·d- 1）,the low,middle,high dose Huoxue Qushi formula groups（ 8,16,32 g·kg- 1·d- 1）,the model and the normal group. After 4-week intervention,the levels of alanine aminotransferase（ ALT）,aspartate amino transferase（ AST）,triglyceride（ TG）,total cholesterol（ TC）,high density lipoprotein cholesterol（ HDL-C） were determined by a colorimetric method. The levels of fasting blood-glucose（ FBG） and fasting insulin（ FINS） were determined by radioimmunoassay. The levels of tumor necrosis factor-α（ TNF-α）,interleukin（ IL）-10 and IL-6were determined by ELISA. The activity of superoxide dismutase（ SOD） and the content of malonaldehyde（ MDA）were determined. The liver pathology was observed by HE staining and peroxisome proliferator-activated receptor α（ PPARα） mRNA expression of liver homogenate was tested by RT-PCR. The content and distribution of CYP2E1 were tested by the ABC method. Result： Compared to the normal group,the levels of ALT,AST,TG,TC,FBG,FINS,TNF-α and IL-6 increased,the levels of HDL-C and IL-10 decreased,the SOD activity reduced,the contents of MDA and CYP2E1 increased and the PPARα mRNA expression reduced in the model group（ P 〈0. 05）. Compared with model group,all the above indexes and the liver pathology were improved in the Huoxue Qushi formula groups（ P〈 0. 05）. Pathological results showed that the liver tissue injury obviously improved.Conclusion： The Huoxue Qushi formula has certain effect in effectively treating NASH through many ways involved in improving insulin resistance,reducing lipid peroxidation and inhibiting the inflammatory reaction.