摘要
目的 探讨白细胞介素（IL）10基因修饰的树突状细胞（DC）对肝纤维化小鼠的治疗作用及其机制.方法 体外构建IL-10基因修饰的DC（DC-IL-10）,流式细胞术检测DC-IL-10表型及功能的变化.6～8周龄健康雄性BALB/c小鼠按随机数字表法随机分成空白对照组（DC组）、肝纤维化组、阴性对照慢病毒转染树突状细胞（DC-mock）处理组和DC-IL-10治疗组.比较各组小鼠肝功能炎症因子、免疫功能变化,以及肝脏组织形态学情况;实时PCR及免疫印迹法研究DC-IL-10对Wnt/β-连环蛋白信号通路的影响及在肝纤维化中的作用.结果 体外实验显示,组间表面分子（主要组织相容性复合物、CD80、CD86）表达差异有统计学意义（F值分别为14.708、22.503、12.595,P值均〈0.05）;DC-IL-10组明显降低于DC组和DC-mock组（P值均〈0.05）;T淋巴细胞增殖组间比较差异有统计学意义（F=50.295,P〈0.05）,DC-IL-10组T淋巴细胞增殖率明显低于DC组和DC-mock组（P值均〈0.001）.体内实验显示,DC-IL-10治疗组丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平明显低于肝纤维化组（P值均〈0.05）;肿瘤坏死因子（TNF）α、IL-6、IL-1β的表达明显低于肝纤维化组（P值均〈0.05）;调节性T细胞（Treg）比例明显高于肝纤维化组（P〈0.05）;病理显示肝组织损伤轻于肝纤维化组;Wnt3a、α-平滑肌肌动蛋白、β-连环蛋白mRNA表达量及蛋白表达量均明显低于肝纤维化组（P值均〈0.001）.结论DC-IL-10诱导Treg产生免疫耐受,抑制过度的炎症反应,阻断Wnt/β-连环蛋白信号转导通路,进而抑制肝纤维化的进展.
Objective To investigate the therapeutic effects and mechanisms of interleukin-10 （IL-10） gene-modified dendritic cells （DC-IL-10） in mice with liver fibrosis. Methods DC-IL-10 was constructed in vitro, the phenotype and function of which were evaluated by flow cytometry. BALB/c mice were treated with intraperitoneal injection of carbon tetrachloride（CCl4）to establish liver fibrotic model. DC-IL-10 was administrated via tail vein. Animals were divided into 4 groups including normal dendritic cell （DC） control , liver fibrosis only, negative lentiviral transfection DC （DC-mock） and DC-IL-10. Liver function, cytokine secretion, T lymphocyte differentiation and liver histomorphology were tested. Real-time PCR and western blot were used to analyze the effect of DC-IL-10 on Wnt/β-catenin signaling pathway and its role in liver fibrosis. Results When compared with DC control and DC-mock, the expression of DC-IL-10 surface stimulating molecules （major histocompatibity complex-Ⅱ, CD80, CD86） were significantly decreased （F=14.708, 22.503, 12.595, respectively, all P〈0.05）, and DC-IL-10 significantly inhibited T lymphocyte proliferation （F=50.295, P〈0.05）. When compared with liver fibrosis group, serum alanine aminotransferase and aspartate transaminase were decreased in DC-IL-10 treated group （all P〈0.05）, other parameters including inflammatory factors （tumor necrosis factor α, IL-6, IL-1β） reduced （all P〈0.05）, the proportion of regulatory T cells （Treg） increased （F=6.742, P〈0.05）, pathological damage improved, the expression of Wnt3a, α-SMA and β-catenin mRNA and protein significantly reduced in DC-IL-10 treatment group（all P〈0.001）. Conclusions DC-IL-10 induces elevation of Treg for immune tolerance, as well as inhibition of inflammatory response, block of Wnt/β-catenin signaling pathway, which translates into improvement of liver fibrosis.
作者
徐叶进
唐新月
杨敏
张盛果
刘敏慧
郭玉香
卢明芹
Xu Yejin;Tang Xinyue;Yang Min;Zhang Shengguo;Liu Minhui;Guo Yuxiang;Lu Mingqin(Department of Infectious Diseases,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)
出处
《中华内科杂志》
CAS
CSCD
北大核心
2018年第11期835-840,共6页
Chinese Journal of Internal Medicine
基金
金华市科技计划项目（Y20153004）
