残黄片对ANIT诱导黄疸模型大鼠的退黄作用机制分析  

Research on Therapeutic Mechanism of Canhuang Tablets on Jaundiced Rats Induced by ANIT

作　　者：吉日木巴图 谢国明 范娜 周燕萍 韩晋 JIRIMU Ba-tu;XIE Guo-ming;FAN Na;ZHOU Yan-ping;HAN Jin(Academy of Mongolian Medicine,Inner Mongolia Medical University,Hohhot 010110,China;Fifth Medical Center of People′s Liberation Army of China(PLA)General Hospital,Beijing 100039,China)

机构地区：[1]内蒙古医科大学蒙医药研究院,呼和浩特010110 [2]中国人民解放军总医院第五医学中心,北京100039

出　　处：《中国实验方剂学杂志》2020年第17期64-69,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市科技计划“十病十药”研发专项(Z171100001717002,Z131100002513005)。

摘　　要：目的:考察残黄片对α-萘异硫氰酸酯(ANIT)诱导黄疸模型大鼠肝脏法尼醇X受体(FXR),尿苷二磷酸葡萄糖醛酸转移酶1A1(UGT1A1)和多药耐药相关蛋白2(MRP2)mRNA和蛋白表达的影响,探讨其治疗残留黄疸的作用机制。方法:大鼠分为正常组、模型组、残黄片(CHP)组和熊去氧胆酸片(UDCA)组,ANIT灌胃复制黄疸模型,相应药物干预后进行血清总胆红素(TBIL),总胆汁酸(TBA),丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST)和碱性磷酸酶(ALP)的含量检测和肝组织病理学检查,以评价残黄片的治疗作用。应用逆转录-聚合酶链式反应(RT-PCR)和蛋白质免疫印迹试验(Western blot)检测各组大鼠肝组织FXR,UGT1A1,MRP2 mRNA和蛋白的相对表达量。结果:CHP能显著降低由ANIT引起的大鼠血清TBIL,TBA,ALT,AST,ALP的升高,抑制肝脏病理学改变,退黄作用优于UDCA。与正常组比较,ANIT造模后显著抑制大鼠肝组织FXR,UGT1A1,MRP2的mRNA水平(P<0.01)。与模型组比较,CHP和UDCA干预后均显著提升了各蛋白目的基因mRNA水平(P<0.01),在提升胆红素代谢酶UGT1A1的mRNA水平上CHP优于UDCA(P<0.01)。在影响蛋白表达方面,与正常组比较ANIT造模使大鼠FXR表达明显升高(P<0.05),CHP干预有促进FXR表达的趋势,而UDCA未见有促进趋势,但二者无显著差异。在促进胆红素代谢和胆汁排泄方面,ANIT造模使得UGT1A1,MRP2的表达显著降低(P<0.01),而CHP治疗后显著提高了UGT1A1和MRP2蛋白的表达(P<0.01)。在提高胆红素与胆汁酸外排蛋白MRP2的表达上CHP优于UDCA(P<0.01)。结论:CHP退黄作用机制与激活肝脏FXR mRNA的表达,促进胆红素代谢酶UGT1A1和胆汁酸转运体MRP2的mRNA和蛋白表达,提高肝脏对游离胆红素的代谢并促进胆汁酸排出肝脏,缓解胆汁淤积性肝损伤有关。Objective:To explore the therapeutic mechanism of Canhuang tablets on the mRNA and protein expression of farnesoid X receptor(FXR),uridine diphosphate glucuronosyltransferase 1 A1(UGT1A1)and multidrug resistance associated protein 2(MRP2)in the liver of jaundiced rats induced byα-naphthalene isothiocyanate(ANIT).Method:The rats were divided into normal group,model group,Canhuang tablets(CHP)group and ursodeoxycholic acid tablets(UDCA)group.The jaundice model was reproduced by ANIT.After the intervention of the corresponding drugs,the contents of total bilirubin(TBIL),total bile acid(TBA),alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)in serum and the liver histopathology were examined to evaluate the therapeutic effect of CHP.The relative mRNA and protein expressions of FXR,UGT1A1 and MRP2 in rat liver tissues were detected by reverse transcriptionpolymerase chain reaction(RT-PCR)and Western blot.Result:CHP can significantly reduce the increase of TBIL,TBA,ALT,AST and ALP caused by ANIT in rat serum,and inhibit the liver pathological changes,which showed that the removing jaundice effect of CHP was better than UDCA.Compared with the normal group,ANIT significantly inhibited the mRNA levels of FXR,UGT1A1 and MRP2 in rat liver tissues after modeling(P<0.01).Compared with the model group,CHP and UDCA significantly increased the mRNA levels of target genes of each protein after intervention(P<0.01),and CHP was superior to UDCA in improving the mRNA level of bilirubin metabolizing enzyme UGT1A1(P<0.01).In the aspect of affecting protein expression,compared with the normal group,ANIT modeling significantly increased the expression of FXR in rats(P<0.05).CHP intervention showed a tendency to promote the expression of FXR,while UDCA did not,but there was no significant difference between them.In the aspects of promoting bilirubin metabolism and bile excretion,the expressions of UGT1A1 and MRP2 were significantly decreased by ANIT modeling(P<0.01),while the expressions of UGT1A1

关 键 词：残黄片 黄疸 胆汁 胆红素 法尼醇X受体(FXR) 尿苷二磷酸葡萄糖醛酸转移酶1A1(UGT1A1) 多药耐药相关蛋白2(MRP2) 

分 类 号：R22[医药卫生—中医基础理论] R256[医药卫生—中医学]