摘要
目的探讨D-氨基半乳糖(D-galactosamine,D-GalN)和脂多糖(lipopolysaccharide,LPS)诱导的急性肝损伤小鼠外周血辅助性T细胞(helper T cell,Th17)和调节性T细胞(regulatory T cell,Treg)亚群及相关细胞因子水平的变化。方法36只无特定病原体(specefic pathogen free,SPF)级雄性BALB/c小鼠随机分为对照组和模型组。模型组小鼠腹腔注射D-GalN和LPS溶液建立急性肝损伤模型,对照组则注射等体积的生理盐水。采用酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)法检测小鼠血清天冬氨酸转氨酶(aspartate transaminase,AST)和谷丙转氨酶(alanine transaminase,ALT)水平,并联合HE染色法检测小鼠肝组织损伤情况;采用流式细胞仪检测小鼠外周血Th17、Treg细胞亚群水平;采用ELISA检测小鼠血清白细胞介素(interleukin,IL)-17、IL-6、IL-23、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL-10、转化生长因子-β(transforming growth factor-β,TGF-β)等细胞因子水平。结果与对照组相比,模型组小鼠外周血AST和ALT水平均显著升高(t值分别为5.39,4.92,P值均<0.05),病理学染色显示小鼠肝组织损伤严重,表明小鼠急性肝损伤模型构建成功;模型组小鼠外周血Th17细胞明显升高,Treg细胞显著下降,Th17/Treg细胞比率明显上升(t值分别为3.87,3.25,5.50,P值均<0.05);此外,模型组小鼠血清中IL-17、IL-6、IL-23及TNF-α的水平明显升高,而IL-10、TGF-β的水平则显著下降,差异有统计学意义(t值分别为4.33,5.60,4.05,3.68,3.12,3.03,P值均<0.05)。结论D-GalN/LPS诱导的急性肝损伤小鼠外周血Th17/Treg细胞平衡严重失调,由其分泌的或与其增殖发育相关的细胞因子表达异常,可能在急性肝损伤的发生发展中起着重要作用。
Objective To investigate the changes of helper T cell 17(Th17)and regulatory T cell(Treg)subsets and cytokine levels in peripheral blood of mice with acute liver injury induced by D-galactosamine(D-GalN)and lipopolysaccharide(LPS).Methods Total of 36 specefic pathogen free(SPF)male BALB/c mice were randomly divided into control group and model group.The model group were injected with D-GalN and LPS solution to establish mouse model with acute liver injury,while the control mice were injected with equal volume of saline.Serum aspartate transaminase(AST)and alanine transaminase(ALT)were detected by enzyme linked immunosorbent assay(ELISA).The liver injury of mice was detected by HE staining.Th17 and Treg in peripheral were detected by flow cytometry.Serum interleukin(IL)-17,IL-6,IL-23,tumor necrosis factor-α(TNF-α),IL-10,and transforming growth factor-β(TGF-β)were analyzed with enzyme linked immunosorbent assay(ELISA).Results Compared with control mice,serum AST and ALT of the model mice were significantly increased(t=5.39,4.92,P<0.05).Pathological staining showed that the liver tissue of model mice was severely damaged,indicating that the mouse model of acute liver injury was successfully generated.Th17 cells in model mice were significantly increased,the Treg cells were significantly decreased,whereas the Th17/Treg cell ratio increased significantly(t values were 3.87,3.25 and 5.50 respectively,all P values<0.05).IL-17,IL-6,TNF-α,and IL-23 in model mice were significantly increased,while IL-10 and TGF-βwere significantly decreased(t values were 4.33,5.60,4.05,3.68,3.12 and 3.03 respectively,all P values<0.05).Conclusion Th17/Treg cells and related cytokines in peripheral blood of mice with acute liver injury induced by D-GalN/LPS are seriously imbalanced,which may play an important role in the development of acute liver injury.
作者
田园
屠昌明
Tian Yuan;Tu Changming(Department of Clinical Laboratory,Joint Hospital of Chinese People's Liberation Army,901 Hospital,Hefei 230031,China)
出处
《国际免疫学杂志》
CAS
2019年第3期274-279,共6页
International Journal of Immunology