西罗莫司促进小鼠心脏移植模型中调节性T细胞的分化和增殖
Sirolimus promotes differentiation and proliferation of regulatory T cells in mouse heart transplantation model
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摘要:目的 探讨西罗莫司(SRL)对小鼠异位心脏移植模型的移植物存活时间以及脾脏中调节性T细胞(Treg)分化和增殖的影响。方法 应用Cuff法建立雄性BALB/c→C57BL/6小鼠颈部异位心脏移植模型。术后随机分为3组各10只受体:对照组术后不予特殊药物治疗, SRL组术后1~14 d予SRL 10 mg/(kg·d)灌胃, 环孢素(CsA)组术后1~14 d予CsA 30 mg/(kg·d)灌胃。记录移植心脏存活时间, 于移植心停搏或术后14 d取脾, 分离单个核细胞, 上流式细胞仪检测CD4+CD25+Treg占CD4+T细胞比例(CD4+CD25+Treg%), 采用逆转录-聚合酶链反应(RT-PCR)法半定量检测Foxp3信使核糖核酸(mRNA)表达水平。结果 与对照组比较, CsA组和SRL组均明显延长小鼠移植心的生存时间(均为P < 0.01), 而两者之间比较差异无统计学意义(P>0.05)。与对照组比较, CsA组脾脏中CD4+CD25+Treg%明显降低, 而SRL组则明显升高(均为P < 0.01), 后两组比较差异有统计学意义(P < 0.01)。SRL组脾脏T细胞的Foxp3 mRNA表达明显高于对照组和CsA组, 其中对照组亦明显高于CsA组(均为P < 0.01)。结论 在小鼠心脏移植模型中, SRL明显延长移植物的存活期, 促进CD4+CD25+Treg的增殖与生长, 有利于免疫耐受的形成。Abstract:Objective To investigate the impacts of sirolimus (SRL) on the survival time of graft and the differentiation and proliferation of regulatory T cell (Treg) of spleen in mouse heterotopic heart transplantation model.Methods Male BALB/c→C57BL/6 mice cervical heterotopic heart transplantation model was established by Cuff method. The mice were divided into 3 groups randomly with 10 mice in each group. The control group received no treatment of special medicine after operation. Mice in SRL group were gavaged with SRL 10 mg/(kg·d) at 1-14 d after operation. Mice in ciclosporin (CsA) group were gavaged with CsA 30 mg/(kg·d)at 1-14 d after operation. The survival time of cardiac grafts were recorded. The spleen was procured after asystole of cardiac graft or 14 d after operation. Mononuclear cells were isolated and the proportion of CD4+CD25+Treg in CD4+ T cell(CD4+ CD25+ Treg%) were detected by flow cytometry and reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of Foxp3 messenger ribonucleic acid (mRNA) semi-quantitatively.Results Compared with the control group, the survival time of cardiac grafts prolonged significantly in SRL and CsA group (all in P < 0.01), but no significant difference was observed between SRL and CsA group(P> 0.05). Compared with the control group, CD4+CD25+ Treg% significantly decreased in the spleen of CsA group and significantly increased in SRL group (all in P < 0.01). And significant difference was observed between SRL and CsA group (P < 0.01). Expression of Foxp3 mRNA of T lymphocyte in the spleen of SRL group was significantly higher than those in control and CsA group (P < 0.01). And expression of Foxp3 mRNA in control group was significantly higher than that in CsA group (P < 0.01).Conclusions In mouse heart transplantation model, SRL can prolong the survival time of graft and promote the proliferation and growth of CD4+ CD25+ Treg to facilitate the establishment of immune tolerance.