小异源二聚体伴侣受体在淤胆型肝炎模型中的表达及大黄素的干预作用

Expression of Small Heterodimer Partner in Rat Model with Acute Cholestatic Hepatitis and Therapeutic Mechanism of Emodin

目的研究小异源二聚体伴侣受体(small heterodimer partner,SHP)在急性淤胆型肝炎大鼠模型中的表达及大黄素的干预作用。方法 40只Sprague-Dawley(SD)大鼠随机分为4组:正常对照组,模型组,大黄素组,熊去氧胆酸(UDCA)组,每组10只。除正常对照组外,其余3组给予α-异硫氰酸萘酯(alphanaphthylisothiocyanate,ANIT)50 mg·kg^(-1)一次灌胃,建立大鼠急性淤胆型肝炎动物模型,并给予相应的药物干预,造模后48 h留取标本,以实时荧光定量PCR法及Western blot法分别检测肝组织中SHPm RNA表达及蛋白表达的变化;全自动生化分析仪检测血清总胆红素(total bilirubin,TB)、直接胆红素(direct bilirubin,DB)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、胆汁酸(total bile acid,TBA)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)、γ-谷氨酰转移酶(gamma glutamine transferase,GGT)的水平;肝组织切片、HE染色后显微镜下观察肝组织病理学改变。结果模型组肝组织中SHP m RNA及蛋白表达水平分别为0.559±0.194、0.313±0.087,均低于正常对照组(P<0.05),大黄素组肝组织中SHP m RNA及蛋白表达水平分别为0.892±0.390、0.706±0.193,均高于模型组(P<0.05);大黄素组血清TB、DB、ALT、TBA、AST、ALP均明显低于模型组(P<0.05),且大黄素组TB、DB、ALT、TBA、AST、ALP明显低于熊去氧胆酸组(P<0.05);大黄素组肝组织病理改变较模型组明显减轻,熊去氧胆酸组肝组织病理改变亦较模型组明显减轻,但程度重于大黄素组。结论大黄素可升高ANIT诱导的大鼠急性淤胆型肝炎肝组织中SHP m RNA及蛋白表达水平。大黄素可降低血清中TB、DB、ALT、TBA、AST、ALP水平及减轻肝组织病理学损害,疗效优于熊去氧胆酸,其作用机制与促进SHP表达有关。

Objective To explore the expression of small heterodimer partner（SHP）in rat model with acute cholestatic hepatitis and to investigate the therapeutic mechanism of emodin. Methods A total of 40 Sprague-Dawley（SD）rats were randomly divided into 4 groups,namely normal group,model group,emodin group and ursodeoxycholic acid group,10 rats in each group. Except for the normal group,rats in other three groups were given intragastric gavage of alpha-naphthylisothiocyanate（ANIT）50 mg·kg^（-1） at one time to induce acute cholestatic hepatitis,and then were given normal saline,emodin,ursodeoxycholic acid respectively according to the grouping. Forty-eight hours after the model establishment,blood samples were collected from abdominal aorta to examine the total bilirubin（TB）,direct bilirubin（DB）,alanine aminotransferase（ALT）,total bile acid（TBA）,aspartate aminotransferase（AST）,alkaline phosphatase（ALP）,and gamma glutamine transferase（GGT）with automatic biochemical analyzer. Real-time PCR and Western blot methods were used to detect the m RNA and protein expression of SHP in the hepatic tissue. Results The SHP m RNA and protein expression levels of the model group were 0.559±0.194,0.313±0.087 respectively,significantly lower than those of the normal group（P 0.05）. The SHP m RNA and protein expression levels of the emodin group were 0.892±0.390,0.706±0.193 respectively,significantly higher than those of the model group（P 0.05）. The serum levels of TB,DB,ALT,TBA,AST,and ALP of the emodin group were all significantly lower than those of the model group（P 0.05）. The serum levels of TB,DB,ALT,TBA,AST,ALP of the emodin group were all significantly lower than those of the ursodeoxycholic acid group（P 0.05）. Pathdogical changes of hepatic tissues of emodin group and ANIT group were improved compared with the model group,and the improvement was more obvious in emodin goup. Conclusion The decreased SHP m RNA and protein levels are shown in hepatic tissue of acute cholestatic hepatitis rats,and emodin has a notable effect on decreasing serum TB,DB,ALT,TBA,AST,ALP levels and on relieving pathological changes of hepatic tissue of rats with ANIT-induced cholestatic hepatitis. Emodin has better effects than ursodeoxycholic acid. And the therapeutic mechanism of emodin is related with the enhancement of SHP expression.