人胎盘MSCs对异基因小鼠皮肤移植排斥的免疫调节作用  

IMMUNOMODULATORY EFFECTS OF HUMAN PLACENTAL-DERIVED MESENCHYMAL STEM CELLS ON IMMUNE REJECTION IN MOUSE ALLOGENEIC SKIN TRANSPLANTATION

作　　者：王琼[1] 刘婷[2] 张耀林[1] 陈冬梅[2] 王立斌[2] 李玉奎[2] 魏军[2] 

机构地区：[1]宁夏医科大学检验学院,银川750004 [2]宁夏医科大学总医院干细胞研究所

出　　处：《中国修复重建外科杂志》2013年第7期775-780,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：宁夏科技攻关项目(2011);宁夏自然科学基金资助项目(NZ11262)~~

摘　　要：目的观察人胎盘MSCs(placental-derived MSCs,PMSCs)对异基因小鼠皮肤移植的影响。方法取自愿捐赠人胎盘胎儿侧组织,分离培养PMSCs,并以第3代细胞进行实验。选择30只6～8周龄C57BL/6小鼠作为受体,背部制作直径12 mm圆形皮肤缺损;以1～2日龄Vr∶CD1(ICR)乳鼠皮肤作为供体,建立小鼠异基因皮肤移植排斥模型。将30只受体小鼠随机分为3组,每组10只。A组为自体皮肤移植组,B组为同种异体皮肤移植+PBS尾静脉注射组,C组为同种异体皮肤移植+人PMSCs(1×105个/只)尾静脉注射组。观察各组移植皮片成活情况,检测术后7 d受体小鼠血液白细胞数量、腹腔液巨噬细胞活化率,采用ELISA、RT-PCR法检测受体小鼠血液、脾脏中IL-4、IL-17及IFN-γ表达量的变化。结果 A、B、C组移植皮片成活时间分别为(58.33±4.04)、(3.80±0.92)、(6.80±0.82)d,A组明显优于B、C组,C组优于B组,差异均有统计学意义(P<0.05)。术后7 d,A、B、C组小鼠血液白细胞计数分别为(6.32±0.45)×109/L、(7.45±0.52)×109/L、(6.35±0.39)×109/L,A、C组显著低于B组(P<0.05),A、C组间差异无统计学意义(P>0.05)。A、B、C组小鼠巨噬细胞活化率分别为6.87%±2.40%、7.84%±0.44%、15.98%±2.87%,C组显著高于A、B组(P<0.01)。ELISA检测示,3组血液中IL-4含量差异无统计学意义(P>0.05);C组IL-17和IFN-γ含量显著低于B组(P<0.05);B组IFN-γ含量明显高于A组(P<0.05);其余各指标组间比较差异均无统计学意义(P>0.05)。RT-PCR检测示,B、C组IL-4、IL-17和IFN-γmRNA相对表达量与A组比较差异均有统计学意义(P<0.05);C组IL-4和IFN-γmRNA相对表达量明显低于B组(P<0.05),IL-17 mRNA相对表达量与B组比较差异无统计学意义(P>0.05)。结论异基因小鼠皮肤移植后尾静脉注射人PMSCs可抑制免疫排斥反应,其机制可能与减少IL-17及IFN-γ等相关细胞因子分泌有关。Objective To investigate the effect of human placental-derived mesenchymal stem cells （PMSCs） on immunological rejection in mouse allogeneic skin transplantation. Methods The placenta fetal tissues from voluntary donors were used to isolate and culture the PMSCs, and the 3rd passage PMSCs were used in the experiment. Thirty Vr ∶ CD1 （ICR） mice at age of 1-2 days were used as skin donors for allogeneic skin transplantation. Thirty C57BL/6 mice at age of 6-8 weeks as recipients were made back skin defect of 12 mm in diameter and were randomly divided into 3 groups （n=10）： group A, autograft; group B, allogeneic graft ＋ PBS tail vein injection; and group C, allogeneic graft ＋ human PMSCs （1 × 105 cells/mouse） tail vein injection. The flap survival was observed. At 7 days after skin transplantation, blood leukocyte counting, abdominal fluid macrophage activation, and the expression levels of interleukin 4 （IL-4）, interleukin 17 （IL-17）, and interferon γ （INF-γ） in blood and spleen were detected by ELISA and RT-PCR, respectively. Results The flap survival time was significantly longer in group A [（58.33 ± 4.04） days] than in groups B and C [（3.80 ± 0.92） days and （6.80 ± 0.82） days] （P 〈 0.05）, and in group C than in group B （P 〈 0.05）. At 7 days after transplantation, the blood leukocyte number was （6.32 ± 0.45） × 109/L in group A, （7.45 ± 0.52） × 109/L in group B, and （6.35 ± 0.39）× 109/ L in group C, and it was significantly more in group B than in groups A and C （P 〈 0.05）. The macrophage activation rate of the abdominal fluid was 6.87% ± 2.40% in group A, 7.84% ± 0.44% in group B, and 15.98% ± 2.87% in group C; group C was significantly higher than groups A and B （P 〈 0.01）. ELISA results showed that there was no significant difference in the concentrations of IL-4 among 3 groups （P 〉 0.05）. Compared with group B, the concentrations of IL-17 and IFN-γ were significantly reduced in group C （P 〈 0.05）, wh

关 键 词：胎盘MSCs 皮肤移植 免疫排斥 异基因 ICR乳鼠 小鼠 

分 类 号：R622[医药卫生—整形外科]