摘要
目的探讨大鼠免疫性肝损伤模型建立的方法以及辅助性T淋巴细胞(Th)17/调节性T淋巴细胞(Treg)失衡在免疫性肝炎大鼠模型中的变化特点。方法采用随机数字表法将30只雌性Wistar大鼠分为3组,每组10只,分别是急性免疫性肝损伤模型(AC)组、慢性免疫性肝损伤模型(CC)组和健康对照(HC)组。AC组和CC组分别通过尾静脉注射α-galcer和刀豆蛋白A(ConA)建立,HC组注射等量的生理盐水。建模成功后检测血清转氨酶ALT、AST等;HE染色观察大鼠肝脏病理改变;免疫组化法检测肝脏中Foxp3、RORγt蛋白表达。外周血和脾脏悬液中Th17、Treg细胞频率变化采用流式细胞分析术检测并计算Th17/Treg比值。计量资料多组间比较采用单因素方差分析,进一步两两比较用SNK-q检验。结果与HC组比较,AC组、CC组血清中ALT、AST、TBil均升高,Alb水平降低,差异均有统计学意义(F值分别为14.782、20.765、16.088、74.181,P值均<0.001)。HE染色,与HC组相比,AC组、CC组分别出现急慢性肝细胞损伤改变。与HC组相比,AC组Th17细胞频率及蛋白(RORγt)水平均显著升高(P值均<0.05),Treg细胞频率亦升高(P<0.05),Th17/Treg比值也显著上升(P<0.05);CC组Th17细胞频率、Treg细胞频率均显著升高,Th17/Treg比值明显下降(P值均<0.05)。结论α-galce及ConA均可成功建立免疫性肝损伤模型,均未出现死亡,肝组织学及血清生化学变化符合急、慢性免疫性肝炎特点。Th17/Treg失衡在炎症反应、组织损伤中发挥了重要作用,且与其免疫状态和病情进展密切相关。
Objective To investigate the methods for establishing a rat model of immunological liver injury and the change in T helper 17 cell(Th17)/regulatory T cell(Treg)imbalance in rats with immunological liver injury.Methods A total of 30 female Wistar rats were divided into acute immunological liver injury group(AC group),chronic immunological liver injury group(CC group),and healthy control group(HC group)using a random number table,with 10 rats in each group.The rats in the AC group and the CC group were given tail vein injection ofα-galactosylceramide(α-GalCer)and concanavalin A(ConA)to establish a rat model,and those in the HC group were given injection of an equal volume of normal saline.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured after modeling,HE staining was used to observe liver pathological changes,and immunohistochemistry was used to measure the protein expression of Foxp3 and RORγt in the liver.Flow cytometry was used to measure the changes in the frequencies of Th17 and Treg cells in peripheral blood and spleen suspension,and Th17/Treg ratio was calculated.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups.Results Compared with the HC group,the AC group and the CC group had significant increases in the serum levels of ALT,AST,and total bilirubin and a significant reduction in the level of albumin(F=14.782,20.765,16.088,and 74.181,all P<0.001).HE staining showed that compared with the HC group,the AC group and the CC group had the changes of acute and chronic hepatocyte injury.Compared with HC group,the AC group had significant increases in the frequency of Th17 cells,the protein expression of RORγt(all P<0.05),the frequency of Treg cells(P<0.05),and Th17/Treg ratio(P<0.05).Compared with the HC group,the CC group had significant increases in the frequencies of Th17 and Treg cells and a significant reduction in Th17/Treg ratio(all P<0.05).Conclusion Bothα-GalCer and ConA can be used to successfully establish a model of immunological liver injury without deaths,and the changes in liver histology and serum biochemistry are consistent with acute and chronic immune hepatitis.Th17/Treg imbalance plays an important role in inflammatory response and tis-sue damage and is closely associated with immune status and disease progression.
作者
薛苗
毛小荣
陈红
XUE Miao;MAO Xiaorong;CHEN Hong(Department of Rheumatology and Immunology,The First Hospital of Lanzhou University,Lanzhou 730000,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2020年第1期149-152,共4页
Journal of Clinical Hepatology
基金
甘肃省自然科学基金(1606RJZA127)
