FK506对肝细胞癌模型大鼠肝移植术后免疫调节的影响  

The Effect of FK506 on Immune Regulation after Liver Transplantation in Rats with Hepatocellular Carcinoma

作　　者：王元元 邓卫平 彭吉霞 吴胜英 WANG Yuan-yuan;DENG Wei-ping;PENG Ji-xia;WU Sheng-ying(Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China;Functional laboratory,Hubei University of Medicine,Shiyan,Hubei 442000,China;School of Biomedical Engineering,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]十堰市太和医院·湖北医药学院附属医院,湖北十堰442000 [2]湖北医药学院基础医学院机能实验室,湖北十堰442000 [3]湖北医药学院基础医学院病理生理教研室,湖北十堰442000

出　　处：《湖北医药学院学报》2019年第5期430-434,439,共6页Journal of Hubei University of Medicine

基　　金：湖北省教育厅指导性项目(B2016125)

摘　　要：目的:观察他克莫司(FK506)对大鼠肝细胞癌模型肝移植术术后免疫功能的影响。方法:雄性Lewis大鼠76只,DA大鼠48只。选取其中60只Lewis大鼠用二乙基亚硝胺灌胃构建肝癌模型,14周后将48只正常DA大鼠肝脏移植于造模后存活的48只肝癌模型大鼠,术后随机分为模型组,FK506高、中、低剂量组(n=12);另16只正常雄性Lewis大鼠作为正常对照组。术后FK506治疗组分别以1、0.5和0.25 mg/kg剂量灌胃4周,正常对照组和模型组给予0.9%氯化钠溶液。于治疗2周和4周时采用ELISA法分别检测外周血一氧化氮(nitric oxide,NO)及肝组织白介素-2β(Interleukin-2β,IL-2β)、IL-4β、IL-12β、干扰素-γ(Interferon-γ,IFN-γ)、转化生长因子-β1(Transforming growth factor beta,TGF-β1)及肿瘤坏死因子(Tumor necrosis factor,TNF-α)水平;采用流式细胞仪检测Treg、CD4+T、CD8+T细胞数;RT-PCR方法检测肝脏诱导型一氧化氮合酶(Inducible nitric oxide synthase,iNOS)基因表达。结果:FK506中、高剂量组IL-2β、IL-4β、IL-12β、NO、IFN-γ、TGF-β1、TNF-α、CD4+T%、CD8+T%在治疗2周和4周时均明显降低,iNOS mRNA低表达,与模型组比较差异有统计学意义(P<0.05),治疗4周时与同组治疗2周时比较差异有统计学意义(P<0.05)。结论:FK506对二乙基亚硝胺诱导肝细胞癌模型大鼠原位肝移植术后急性排异反应具有明显的抑制作用,这一作用主要是通过调节Treg类细胞因子及NO信号通路蛋白而实现。Objective To investigate the effect of FK506(tacrolimus) on the immune function after liver transplantation in rats with hepatocellular carcinoma. Methods A total of 76 Lewis rats and 48 DA rats were all male. 60 Lewis rats were treated with diethylnitrosamine to construct a liver cancer model. After 14 weeks, 48 normal DA rats were transplanted into 48 surviving liver cancer model rats. After operation, they were randomly divided into model group, FK506 high, medium and low dose groups(n=12);another 16 normal male Lewis rats served as normal control group. The FK506 treatment group was intragastrically administered for 1 week at 1, 0.5 and 0.25 mg/kg, respectively. The normal control group and the model group were given 0.9% sodium chloride solution. Peripheral blood nitric oxide(NO) and liver tissue IL-2β, IL-4β, IL-12β, IFN-γ, TGF-β1 and TNF-α were detected by ELISA at 2 and 4 weeks of treatment. Treg was detected by flow cytometry. The number of Treg, CD4+T and CD8+ T cells was detected by flow cytometry. The expression of inducible nitric oxide synthase(iNOS) gene was detected by RT-PCR. Results IL-2β, IL-4β, IL-12β, NO, IFN-γ, TGF-β1, TNF-α, CD4+T%, and CD8+T% in the medium and high dose groups of FK506 were significantly decreased at 2 and 4 weeks of treatment. The expression of iNOS mRNA was significantly lower than that of the model group(P<0.05). There was significant difference between the two groups at the 4 th week(P<0.05). Conclusion FK506 has a significant inhibitory effect on acute rejection after orthotopic liver transplantation in rat models with hepatocellular carcinoma induced by diethylnitrosamine. This effect is mainly achieved by modulating Treg-like cytokines and NO of signaling pathway proteins.

关 键 词：他克莫司 肝细胞癌 原位肝脏移植 Treg类细胞因子 一氧化氮 

分 类 号：R73[医药卫生—肿瘤]