胸腺内注射供体BMSCs对同种异体腹部皮瓣移植排斥的影响  

The Effects of Intrathymic Injection of Donor BMSCs on the Survival of Rat Groin Allograft

作　　者：张栋梁[1] 宋雅娟[1] 苏映军[1] 陈建武[1] 陈晨[1] 丁健科[1] 刘甜[1] 郭树忠[1] 

机构地区：[1]第四军医大学西京医院整形外科研究所,陕西西安710032

出　　处：《现代生物医学进展》2015年第8期1465-1470,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金重点项目(30830102)

摘　　要：目的:将供体骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)胸腺内注射(intrathymic injection,IT)至受体胸腺,探讨其对同种异体腹部皮瓣移植排斥的影响及机制。方法:全骨髓贴壁培养法培养并纯化BN大鼠BMSCs,流式细胞术对其表型进行鉴定。受体Lewis鼠随机分为A组(空白对照组)、B组(BMSCs组)、C组(60Coγ射线4Gy全身照射组)、D组(全身照射+BMSCs组),每组6只。各组大鼠第0天进行同种异体腹部皮瓣移植,A组移植前14天IT PBS,B组移植前14天IT供体BMSCs,C组移植前15天给予60Coγ射线4Gy全身照射,移植前14天IT PBS,D组除移植前15天4Gy全身照射外,移植前14天IT供体BMSCs。大体观察移植皮瓣存活情况并绘制生存曲线,排斥终点流式检测脾细胞调节性T细胞(Treg)比例变化,体外混合淋巴细胞反应检测受体脾淋巴细胞对供体抗原反应性变化。结果:全骨髓贴壁培养法能够很好培养出BMSCs,其P3代流式细胞术表型鉴定结果为CD11b/c、CD45阴性,CD29、CD90阳性。B组的移植皮瓣存活时间与A组相比无明显差异(P>0.05),而D组的移植皮瓣平均存活时间较C组延长3.4天,差异具有统计学意义(P<0.01),脾细胞Treg比例显著增高(P<0.01),脾淋巴细胞对供体抗原反应性显著降低(P<0.05)。结论:IT供体BMSCs联合60Coγ射线4Gy全身照射通过上调受体脾细胞Treg比例能有效降低受体脾淋巴细胞对供体抗原的反应性,显著延长同种异体腹部皮瓣移植物存活时间。Objective： To investigate the effects of intrathymic injection of donor BMSCs（bone marrow mesenchymal stem cells）on the survival of groin allograft in a rat model of groin allotransplantation and its mechanisms. Methods： The BN rat BMSCs were cultured and purified with the application of whole bone marrow adherent culture and the phenotypes were identified by flow cytometry.Twenty four recipient lewis rats were randomly divided into four groups, including group A（control group）, group B（BMSCs group）,group C（4 Gy60Coγ ray total-body irradiation） and group D（4 Gy total-body irradiation plus BMSCs）. Every recipient rat accepted groin allotransplantation on day 0, group A IT（intrathymic injection） PBS on day-14, group B IT BMSCs on day-14, group C accepted 4 Gy60Coγ ray total-body irradiation on day-15 and IT PBS on day-14. In addition to 4 Gy total-body irradiation, group D IT BMSCs on day-14. The mean survival time of groin allograft, change of Treg（regulatory T cells） ratio of spleen cells and MLR（mixed lymphocyte reaction） at the end of rejection were observed. Results： The whole bone marrow adherent culture could effectively culture BMSCs, and the P3 of BMSCs were negative for CD11b/c and CD45 but positive for CD29 and CD90. The mean survival time of groin allograft in group B had no significant difference compared with group A（P〈0.05）, while the groin allograft in group D had significantly longer survival time than that of group C（P〈0.01）. The ratio of Treg in spleen cells was enhanced significantly（P〈0.01）, and in MLR, the reactivity of spleen lymphocytes to donor antigen in group D was significantly lower than those of group C（P〈0.05）. Conclusion： IT donor BMSCs plus 4 Gy total-body irradiation could effectively reduce the reactivity of recipient lymphocyte to donor antigen and prolong the survival time of groin allograft through up regulation of Treg ratio.

关 键 词：胸腺内注射 骨髓间充质干细胞 同种异体腹部皮瓣移植 免疫耐受 

分 类 号：R622[医药卫生—整形外科]