龙胆泻肝丸对胆汁淤积大鼠肝脏多药耐药蛋白及中性粒细胞CD18表达影响的研究  

Effects of Longdan Xiegan Pill on Expression of Multidrug Resistance Protein and Neutrophil CD18 in Cholestatic Rats

作　　者：董伟[1] 梁爱华[1] 李春英[1] 汤喜兰[1] 王金华[1] 薛宝云[1] 王秀坤[1] 

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2011年第21期214-217,共4页Chinese Journal of Experimental Traditional Medical Formulae

摘　　要：目的:从与胆汁分泌有关的转运蛋白及中性粒细胞CD18分子的角度探讨龙胆泻肝丸(LD)清肝胆,利湿热的可能作用机制。方法:将20只大鼠分为4组:对照组,模型组,LD 5.0,2.5 g.kg-1(按生药计算)组。给药组每日ig给药1次,连续8 d,对照组和模型组ig等体积的蒸馏水。其中第5天给药1 h后,除对照组ig花生油外,其他各组大鼠按100 mg.kg-1 1次性igα-萘异硫氰酸酯(ANIT)花生油。第8天摘取大鼠肝脏,提取肝脏总RNA,RT-PCR检测多药耐药蛋白MDR1a,MDR1b,MDR2和多药耐药相关蛋白MRP1 mRNA,MRP2 mRNA的表达;眼眶静脉取抗凝血,流式细胞仪测定中性粒细胞CD18表达,血细胞计数仪测定白细胞及粒细胞总数。结果:模型组大鼠肝组织MDR1a,MDR1b mRNA表达显著增加(P<0.05,P<0.01),MDR2 mRNA,MRP2 mRNA分别有表达增加或降低的趋势,MRP1 mRNA的表达无明显变化。LD 5.0,2.5 g.kg-1对胆汁淤积大鼠的肝组织MDR1a,MDR1b,MDR2,MRP1,MRP2 mRNA表达均无明显影响。LD各给药组白细胞总数以及粒细胞总数有下降趋势,CD18荧光强度明显低于模型组(P<0.05或P<0.01)。结论:龙胆泻肝丸对ANIT诱导的胆汁淤积(类肝胆湿热证)具有利胆保肝作用,其作用机制主要与其抑制白细胞黏附,降低胆管炎症反应,减轻胆管损伤,从而有利于胆汁排泄有关,而与胆管上皮的转运体蛋白分子MDR,MRP关系可能不大。Objective：To observe the effect of Longdan Xiegan pill（LD） on expression of multidrug resistance protein and neutrophil CD18 and tclarify the possible mechanism of LD to clear dampness-heat of liver and gallbladder.Method：Twenty healthy Wistar rats were randomly divided into 4 groups：control group,ANIT-treated model group,LD 5.0 g·kg-1group and LD 2.5 g·kg-1group.LD groups were administered orally for 8 days,while control group and model group were administered orally with equal distilled water,once daily.After the last administration for 1 h,model group and LD groups were treated with 100 mg·kg-1 ANIT,while control group was administered with equal peanut oil.After administration of ANIT 72 hours,The liver total RNA of all rats wwas extracted and the levels of MDR1a,MDR1b,MDR2,MRP1 and MRP2 mRNA in liver tissue were detected by reverse transcription polymerase chain reaction（RT-PCR）.The expression of CD18 was measured by using flow cytometer.Total leukocytes and granular leukocytes were counted by using microcomputer controlled hematology analyzer.Result： The expression of MDR1a,MDR1b mRNA in model group wwas significantly higher than that of control group（P〈0.05,P〈0.01）.The expression of MDR2,MRP1,MRP2 mRNA was not obviously changed.LD 5.0,2.5 g·kg-1 group had no obvious effects on expression of multidrug resistance protein in cholestatic rats.LD groups could reduce total leukocytes granular leukocytes and CD18 fluorescence intensity.Conclusion：LD has protection in ANIT-induced cholestatic rats,which is similar to the syndrome of dampness-heat of liver and gallbladder.The protection mechanism maybe mainly related to inhibiting endothelial-leukocyte adhesion,inhibiting inflammatory responses in bile ducts,alleviating biliary injuries,increasing bile flow of intrahepatic bile ducts,not related to the expresion of MRP1 and MRP2 mRNA in biliary epithelial cells.

关 键 词：龙胆泻肝丸 胆汁淤积 多药耐药蛋白 多药耐药相关蛋白 CD18 

分 类 号：R285.5[医药卫生—中药学]