南方医科大学学报2017，Vol.37Issue(4)：451-459,9.DOI:10.3969/j.issn.1673-4254.2017.04.05
阻断Kupffer细胞TIM-4功能对小鼠肝移植排斥反应的影响
Effect of inhibiting TIM-4 function in Kupffer cells on liver graft rejection in mice
摘要
Abstract
Objective To investigate the effects of inhibiting TIM-4 function in Kupffer cells (KCs) on liver graft rejection in mice and explore the underlying mechanism. Methods Mouse models of orthotopic liver transplantation were treated with a control mAb group and TIM-4 mAb. The activated KCs were assayed with immunohistochemistry after operation. The expression of TIM-4 in KCs were assayed with Western blotting and RT-PCR and the levels of AST, ALT, TBIL, TNF-α, IFN-γand CCL2 were assayed detected. The expression of TIM-4 in KCs was observed with laser confocal microscopy. HE staining was used to observe the microstructure of the liver tissues, and the number of CD25 +Foxp3 +T cells was determined using with flow cytometry; the proteins levels of p-P65and p-P38 were assayed with Western blotting. The donor mice were treated with clodronate liposomes to destroy the KCs in the liver before transplantation, and the liver grafts were examined for graft rejection. Results The number of activated KCs in the liver graft increased progressively over time. Compared with the sham-operated group, the liver graft showed significantly increased TIM-4 protein and mRNA levels at 1, 3, and 7 days after transplantation (P<0.05) and increased levels of AST, ALT, TBIL, TNF-α, IFN-γ and CCL2 at 7 days (P<0.05). The graft in TIM-4 mAb group showed mild pathological changes with a mean RAI score of 2.67 ± 0.75, which was significantly lower than that in control mAb group (P<0.05). The mean survival time of the recipient mice was 53.8±6.4 days in TIM-4 mAb group, significantly longer than that in the control mAB group (14.5±2.9 days, P<0.05). Donor treatment with clodronate liposomes resulted in comparable RAI scores in TIM-4 mAb and control mAb groups (8.01±0.64 vs 7.93±0.56, P>0.05). Theprotein levels of p-P65 and p-P38 in TIM-4 mAb group were significantly lower than those in control mAb group (P<0.05), and CD25+Foxp3+T cells in the liver graft increased significantly in TIM-4 mAb group. Conclusion Inhibition of TIM-4 function in KCs reduces the production of inflammatory factors after liver transplantation possibly by inhibiting the NF-κB and MAPK signaling pathways and promoting the proliferation of Foxp3+Treg cells to induce allograft tolerance.关键词
T细胞免疫球蛋白黏蛋白4/Kupffer细胞/肝移植/急性排斥反应Key words
T-cell immunoglobulin and mucin-domain-containing molecule 4/Kupffer cells/liver transplantation/acute rejection引用本文复制引用
李学强,李旭宏,段世刚,徐雪松,刘一鸣,李金政,龚建平,吴皓..阻断Kupffer细胞TIM-4功能对小鼠肝移植排斥反应的影响[J].南方医科大学学报,2017,37(4):451-459,9.基金项目
国家自然科学基金(81370580) （81370580）
国家自然科学基金青年基金(81300364) （81300364）
重庆市卫生计生委医学科研计划(2015MSXM123, 2015ZDXM029) Supported by National Natural Science Foundation of China (81370580) and the National Science Foundation for Distinguished Young Scholars of China (81300364). （2015MSXM123, 2015ZDXM029）
