葛花解酒涤脂汤对酒精性脂肪肝小鼠肝脏核受体PXR,CAR表达的影响

Intervention Effect of Gehua Jiejiu Dizhi Decoction on Alcoholic Fatty Liver and Liver PXR,CAR Expression in Mice

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 explore the effect of Gehua Jiejiu Dizhi decoction on the liver pregnane X receptor（PXR）,constitutive androstane receptor（CAR） and their target genes CYP3A11,CYP3A25,CYP2B9 and CYP2B10 in mice with alcoholic fatty liver. Method： Twenty-nine C57 BL /6J mice were randomly divided into four groups,namely normal control group,model group,and Gehua Jiejiu Dizhi decoction high-dose and low-dose groups. At first,the alcoholic fatty liver model in mice was prepared by using National Institute on Alcohol Abuse and Alcoholism（NIAAA） method. Then,Gehua Jiejiu Dizhi decoction was given orally at doses of 4. 9,2. 45 g·kg- 1·d- 1by gavage（once a day for up to 9 days）. Serum and liver tissue samples were collected for biochemical and histopathological examination of liver injury. Enzyme linked immunosorbent assay（ELISA） was performed to detect free cholesterol（FCHOL） and triglyceride（TG） levels. HE staining was used to detect liver injury. Realtime PCR and Western blot were used to examine the expressions of PXR,CAR and their target genes CYP3A11,CYP3A25,CYP2B9 and CYP2B10 in mice liver. Result： Compared with model group, hepatic steatosis significantly reduced in Gehua Jiejiu Dizhi decoction-treated mice in a dose-dependent manner. Compared with model group and blank group,expressions of PXR and CYP3A25 significantly increased in Gehua Jiejiu Dizhi decoction-treated mice（P〈 0. 01）. CAR mRNA level significantly rose（P 〈0. 05）. Compared with normal group,FCHOL and TG levels significantly increased in model group（P〈 0. 01）. Conclusion： Gehua Jiejiu Dizhi decoction could significantly ameliorate AFLD in mice,which was possibly correlated with the activation of PXR and its regulatory genes CYP3A25.