白细胞介素-18反义寡脱氧核苷酸对同种异体部分移植肝再生的促进作用

Interleukin-18 antisense ollgodeoxynucleotide promotes hepatocyte regeneration of partial liver allograft

目的 探讨白细胞介素(IL)-18硫代修饰反义寡脱氧核苷酸(ASPODN)对同种异体部分移植肝再生的促进作用。 方法 将供体SD大鼠与受体LEW大鼠随机分为50％部分肝移植组(PLT组)、部分肝移植IL-18 ASPODN治疗组(IL-18 ASPODN组)及部分肝移植IL-18硫代修饰正义寡脱氧核苷酸治疗组(IL-18 SPODN组),每组供、受体大鼠各30只。检测移植肝肝细胞BrdU标记变化;检测移植肝IL-18蛋白和γ干扰素(INF-γ)mRNA表达以及受体血清IFN-γ水平。 结果 移植术后72 h,3组移植肝肝细胞再生均达到高峰。IL-18 ASPODN组移植肝肝细胞BrdU高峰标记指数(58.3％±7.5％)明显高于PLT组(31.6％±6.7％)及IL-18 SPODN组[(33.4±5.5％),t=6.503、6.558,P<0.001];移植术后48、72和96 hIL-18 ASPODN组移植肝内IL-18蛋白和IFN-γ mRNA表达量以及受体血清IFN-γ水平均明显低于PLT组(IL-18蛋白比较:t=2.950,P<0.05;t=5.916、7.947,P<0.001;INF—γ mRNA比较:t=2.558,P<0.05;t=6.292、8.925,P<0.001;IEN-γ水平比较:t=16.998、15.483、54.723,P<0.001)和IL-18SPODN组(IL-18蛋白比较:t=2.845,P<0.05;t=6.062、6.973,P<0.001;INF-γ mRNA比较:t=3.117,P<0.05;t=6.154、8.738,P<0.001;IFN-γ水平比较:t=14.531、18.139、46.924,P<0.001)。结论 IL-18

Objective To study the effect of interleukin (IL)-18 ASPODN on regeneration of allogeneic partial liver graft in rats. Methods Ninety donor SD rats and ninety recipient LEW rats were randomly divided into 3 groups: 50% partial liver transplantation group(PLT group); PLT+IL-18 antisense phosphorothioate oligodeoxynucleotide (ASPODN) treatment group (IL-18 ASPODN group) and PLT+IL-18 SPODN treatment group(IL-18 SPODN group) in which iiposomes encapsulated IL-18 ASPODN or IL-18 SPODN were intravenous injection every day after PLT. BrdU labeling of hepatocytes, expression of IL-18 protein and IFN-γ mRNA in liver graft, and serum level of IFN-γ were measured with immunohistochemistry analysis, Western blotting, semi-quantification RT-PCR , and ELISA, respectively. Results Although regeneration of liver graft from each group peaked 72 h after transplantation, BrdU labeling of hepatocytes in IL-18 ASPODN group (58.3% ± 7.5%) were significantly higher than those of PLT group (31.6% ± 6.7%) (t =6.503, P<0.001) and IL-18 SPODN group (33.4% ± 5.5%)(t=6.558, P<0.001). Expression of IL-18 protein and IFN-γ mRNA in liver graft, and serum level of IFN-γ in IL-18 ASPODN group from 48 h ,72 h and 96 h after transplantation were significantly suppressed compared with PLT group (IL-18 protein: t=2.950, t=5.916, t=7.947, P<0.05, P<0.001; INF-γ mRNA: t=2.558, t=6.292, t=8.925, P<0.05, P<0.001; IFN-γ level: t=16.998, t=15.483, t=54.723, P<0.001) and IL-18 SPODN group (IL-18 protein: t=2.845, t=6.062, t=6.973, P<0.05, P<0.001; INF-γ mRNA: t=3.117, t=6.154, t=8.738, P <0.05, P<0.001; IFN-γ level: t=14.531, t=18.139,t=46.924, P<0.001). Conclusion IL-18 ASPODN could promote hepatocyte regeneration of allogeneic partial liver graft by the suppression of IL-18 and IFN-γ production.