绿原酸对慢性胆汁淤积所致大鼠肝纤维化模型胆管及胶原增生的影响  

Effect of Chlorogenic Acid on the Proliferation of Bile Duct and Collagen in Rats with Liver Fibrosis Induced by Chronic Cholestasis

作　　者：唐丽明[1] 李婉华 宋宁[1] 张桂贤[3] 刘大卫[3] TANG Li-ming;Li Wan-hua;SONG Ning(Nankai Santan Hospital,Tianjin(300193),China)

机构地区：[1]天津市南开区三潭医院中医科,天津300193 [2]天津医科大学研究生院,天津300070 [3]天津市医药科学研究所肿瘤药理、病理研究室,天津300020

出　　处：《中国中西医结合外科杂志》2020年第3期423-428,共6页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

摘　　要：目的:采用α-萘异硫氰酸酯(ANIT)多次灌胃给药制备慢性胆汁淤积肝纤维化大鼠模型,观察绿原酸对该模型胆管及胶原增生的影响,探讨其防治机制。方法:健康成年SPF级Wistar雄性大鼠40只,随机分为5组:对照组、模型组、熊去氧胆酸(UDCA)组、绿原酸低剂量组和绿原酸高剂量组,每组8只。除对照组外,其余4组均以ANIT(80 mg/kg)灌胃5周、每周3次,制备慢性胆汁淤积模型,同时UDCA组、绿原酸低剂量组和绿原酸高剂量组,分别给予UDCA(90mg/Kg)、绿原酸(50 mg/Kg和100 mg/Kg)灌胃,每日1次,连续5周。治疗5周结束后,连同对照组、模型组,共5组动物,麻醉状态下,经腹主动脉取血,全自动生化分析仪检测血清中总胆红素(TBIL)、结合胆红素(DBIL)、间接胆红素(IBIL)以及碱性磷酸酶(ALP)水平。切取部分肝组织,中性甲醛固定,进行HE染色、天狼星红染色,观察评价肝组织胆汁淤积、胆管增生及胶原增生等病理改变情况;另取部分肝组织,匀浆提取总蛋白,Western blot法检测肝星状细胞活化标志分子-α-平滑肌肌动蛋白(α-SMA)表达水平。结果:与正常组比,模型组大鼠血清TBIL、DBIL、IBIL、ALP水平显著升高(均P<0.01);与模型组比,绿原酸高、低剂量组以及UDCA组血清TBIL、DBIL、IBIL水平显著降低(P<0.05),ALP有下降趋势,但差异无显著性(P>0.05)。模型组HE染色显示肝组织慢性炎性细胞浸润明显,汇管区胆管增生、扩张显著;绿原酸高、低剂量组及UDCA组肝慢性炎性反应及汇管区胆管增生反应显著减轻。天狼星红染色显示正常组大鼠肝组织胶原表达稀少,而模型组汇管区胶原表达显著增强,偏振光下以亮红黄色的I型胶原为主;与模型组比,绿原酸高、低剂量组以及UDCA组肝组织胶原染色均显著减轻,偏振光下以呈绿色的III型胶原为主。Western blot法检测显示,模型组大鼠肝组织中α-SMA表达水平显著升高(P<0.01),与模型组比,绿原酸高Objective To observe the effect of chlorogenic acid on the proliferation of bile duct and collagen in rats with chronic cholestasis and to explore the mechanism of prevention and treatment.Methods Forty adult male Wistar rats were randomly divided into five groups:control group,model group,UDCA group,low dose group and high dose group.In addition to the control group,the other four groups were treated with ANIT(80 mg/kg)for 5 weeks and 3 times a week to prepare the chronic cholestasis model.At the same time,UDCA group,low dose group and high dose group were given UDCA(90 mg/kg),chlorogenic acid(50 mg/kg and 100 mg/kg)for 5 weeks.At the end of 5 weeks,5 groups of animals,including control group and model group,were taken blood from abdominal aorta under anesthesia,and the levels of TBIL,DBIL,ibil and ALP in serum were measured by automatic biochemical analyzer.Partial liver tissues were cut,fixed with neutral formaldehyde,stained with he and Sirius red,and the pathological changes of cholestasis,bile duct hyperplasia and collagen hyperplasia were observed and evaluated.The total protein was extracted from other liver tissues,and the expression level ofα-smooth muscle actin(α-SMA)was detected by Western blot.Results compared with the normal group,the levels of TBIL,DBIL,IBIL and ALP in the model group were significantly higher(P<0.01);compared with the model group,the levels of TBIL,DBIL and IBIL in the high and low dose chlorogenic acid group and UDCA group were significantly lower(P<0.05),and ALP had a downward trend,but there was no significant difference(P>0.05).HE staining of the model group showed that the chronic inflammatory cell infiltration in the liver tissue was obvious,the proliferation and expansion of bile duct in the portal area were significant;the chronic inflammatory response and the proliferation of bile duct in the portal area were significantly reduced in the high and low dose groups of chlorogenic acid and UDCA groups.Sirius red staining showed that the expression of collagen in the normal

关 键 词：绿原酸 Α-萘异硫氰酸酯 慢性胆汁淤积 Α-平滑肌肌动蛋白 胶原 

分 类 号：R657.4[医药卫生—外科学]