论文部分内容阅读
目的:探讨葛花解酒涤脂汤对小鼠酒精性脂肪肝(AFLD)的干预作用及对肝组织孕烷受体(PXR),组成性雄甾烷受体(CAR)表达的影响。方法:将29只雄性C57BL/6J小鼠随机分为正常组,模型组,葛花解酒涤脂汤高、低剂量组。采用美国国立卫生研究院酒精滥用和酒精中毒研究所(NIAAA)方法制备AFLD小鼠模型,经鉴定成功后,分别灌胃给予高、低剂量葛花解酒涤脂汤(4.9,2.45 g·kg-1·d-1)。9 d后处死小鼠,收集血清及肝组织样本。酶联免疫吸附法检测血清中游离胆固醇(FCHOL)及甘油三酯(TG)水平,苏木素伊红(HE)染色检测肝组织损伤情况;实时荧光定量PCR及蛋白免疫印迹法(Western blot)分析肝组织PXR,CAR mRNA和蛋白表达,及其调控的细胞色素P450氧化酶(CYP3A11,CYP3A25,CYP2B9,CYP2B10)基因表达水平。结果:与模型组比较,葛花解酒涤脂汤组小鼠肝脏脂肪变性显著减轻,并呈剂量依赖性。与模型组和空白组比较,葛花解酒涤脂汤组可显著上调PXR,CYP3A25表达水平(P<0.01),明显增加CAR mRNA水平(P<0.05)。与正常组比较,模型组血清游离胆固醇及甘油三酯水平水平显著增加(P<0.01)。结论:葛花解酒涤脂汤能够显著改善小鼠AFLD,其作用机制可能与肝脏PXR及其调控基因CYP3A25表达的活化有关。
Objective: To investigate the effect of Gehua Jiejiu Decoction on the mice with alcoholic fatty liver (AFLD) and its effect on the expression of pregnane receptor (PXR) and its constituent androstane receptors (CAR) in liver tissues. Methods: Thirty-nine male C57BL / 6J mice were randomly divided into normal group, model group and Gehua Jiejiu Decoction high and low dose groups. The AFLD mouse model was established by the National Institute of Alcohol Abuse and Alcoholism (NIAAA). After successful identification, high and low doses of Gehua Jiejiu Decoction (4.9, 2.45 g · kg -1 -1 · d-1). Mice were sacrificed 9 days later, and serum and liver tissue samples were collected. Serum levels of free cholesterol (FCHOL) and triglyceride (TG) were detected by enzyme-linked immunosorbent assay (ELISA), liver damage was detected by hematoxylin and eosin (HE) staining, and liver was analyzed by real-time fluorescence quantitative PCR and Western blot The mRNA and protein expression of PXR, CAR and CYP3A11, CYP3A25, CYP2B9 and CYP2B10 were detected by Western blot. Results: Compared with the model group, the mice in the Gehua Jiejiu Decoction group had significantly reduced hepatic steatosis in a dose-dependent manner. Compared with model group and blank group, Gehua Jieji Decoction group could significantly up-regulate the expression of PXR and CYP3A25 (P <0.01) and significantly increase the level of CAR mRNA (P <0.05). Compared with the normal group, serum free cholesterol and triglyceride levels in the model group increased significantly (P <0.01). Conclusion: Gehua Jiejiu Decoction can significantly improve the mouse AFLD, the mechanism may be related to the activation of PXR and its regulatory gene CYP3A25.