退黄汤联合熊去氧胆酸对α-萘异硫氰酸酯诱导的大鼠急性肝损的保护作用  

Research of Combination Protective Effects of UDCA and THT on ANIT-induced Liver Injury in Rats

作　　者：薛川[1] 李绍民[2] 徐方明[3] 丁贤君[3] 黎运呈[3] 张国梁[3] 林志益[3] 李绍佐[3] 张浙恩 曾芳[3] 李世波[3] 

机构地区：[1]佳木斯大学研究生学院,佳木斯黑龙江154007 [2]佳木斯大学第一附属医院感染性疾病科,佳木斯黑龙江154002 [3]浙江省舟山医院感染性疾病科,浙江舟山316004

出　　处：《中华中医药学刊》2013年第2期311-315,I0009,I0010,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药管理局资助项目(2008CB073)

摘　　要：目的:探讨退黄汤(THT)联合熊去氧胆酸(UDCA)对α-异硫氰酸萘酯(ANIT)诱导大鼠肝损过程保护作用及机制。方法:选取Sprague-Dawley(SD)大鼠84只,设立空白组6只,对剩余78只在0 h时间点均以ANIT 100 mg/kg予以灌胃造成急性肝损伤,在肝损后24 h取6只处死留取标本(肝损24 h组),然后分为生理盐水组(18只,9 mL/kg体重的生理盐水灌胃)、UDCA组(18只,20 mg/kg体重的UDCA溶液灌胃)、THT组(18只,20 mL/kg体重的THT灌胃)、UDCA/THT组(18只,20 mg/kg体重的UDCA和20 mL/kg体重的THT联合灌胃),各组分别在肝损造模后48 h、72 h、96 h处死6只大鼠。所有大鼠处死留取血清和肝组织,生化仪检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBI)、总胆汁酸(TBA),酶联免疫吸附法(ELISA)检测血清白介素-10(IL-10)白介素-6(IL-6)肿瘤坏死因子-α(TNF-α),实时荧光定量PCR检测肝组织多药耐药相关蛋白2(Mrp2)mRNA,苏木素-伊红(HE)染色观察肝组织炎症活动度。结果:UDCA和THT在降低血清ALT、AST、TBI、TBA、IL-6、TNF-α和提升血清IL-10及肝组织Mrp2 mRNA均存在明显的主效应(P<0.01),两药在调控血清TBI、IL-6、TNF-α、IL-10及肝脏Mrp2 mRNA均存在明显交互协同作用(P<0.01或P<0.05)。HE染色显示联合用药组96 h肝组织炎性程度轻于单药组及生理盐水组。结论:UDCA和THT联合降低了ANIT诱导的肝损作用可能和两药联合作用调控血清细胞因子及肝脏Mrp2表达有关。Objective：To investigate the protective effects and action mechanisms of combinations of Tuihuangtang （THT） and Ursodeoxycholie Acid（UDCA） on α - naphthylisothi（ANIT） - induced liver injury in rats. Methods ： A total of 84 Sprague - Dawley（SD） rats were selected and 6 rats were set to be normal control group. After ANIT intoxication of the remaining 78 rats,6 rats were set to be 24 h - intoxication group. The remaining 72 rats were divided into 4 groups ： saline water group（ 18 rats,treated with intragastric administration of normal saline solution at a dose 9 mL/kg）, UDCA group （18 rats, treated with intragastric administration of UDCA at a dose of 20 mg/kg）, THT group （18 rats, treated with intragastric administration of THT at a dose of 20 mL/kg ）, UDCA/THT group（ 18 rats, treated with intragastric administration of both UDCA at a dose of 20 mg/kg and THT at a dose of 20 mL/kg ）. In 48,72 and 96 hours after ANIT intoxication in each group, 6 rats were killed. Serum levels of alanine transaminase （ALT）, aspartate transaminase（ AST）, total bilirubin （ TBI）, total bile acid （ TBA）, interleukin - 10 （ IL - 10）, interleukin - 6 （ IL - 6） and tumor necrosis factor - α （ TNF - α） were deteced by biochemical analyzer or enzyme linked immunosorbent assay（ELISA）. The muhidrug resistance associated protein2 （Mrp2）mRNA expression levels and inflammation conditions were deteced by quantitative real -time polymerase chain reaction （ qRT - PCR） and Haematoldin - Eosin （HE） staining respectively. Results ： Either UDCA or THT showed significant main effects on down - regulations of serum levels of ALT, AST, TBI, TBA, IL - 6, TNF -α and up - regulations of serum IL- 10 levels and liver Mrp2 mRNA relative expression levels （P 〈 0.01 ）. Additionally, UDCA and THT showed synergistic effects on regulations of serum levels of TBI, IL- 6, TNF-oL, IL- 10 and liver Mrp2 mRNA expression levels（P 〈 0.01 or P 〈 0.05 ）. HE staining showed

关 键 词：熊去氧胆酸 退黄汤 α-异硫氰酸萘酯 细胞因子 多药耐药相关蛋白2 

分 类 号：R285.5[医药卫生—中药学]