Wang Qiong, Liu Ting, Zhang Yaolin, Chen Dongmei, Wang Libin, Li Yukui, Wei Jun
Department of Medical Laboratory, Ningxia Medical University, Yinchuan Ningxia, 750004, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Jul;27(7):775-80.
To investigate the effect of human placental-derived mesenchymal stem cells (PMSCs) on immunological rejection in mouse allogeneic skin transplantation.
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 x 10(5) 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 gamma (INF-gamma) in blood and spleen were detected by ELISA and RT-PCR, respectively.
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) x 10(9)/L in group A, (7.45 +/- 0.52) x 10(9)/L in group B, and (6.35 +/- 0.39) x 10(9)/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-gamma were significantly reduced in group C (P < 0.05), while the concentration of IFN-gamma was significantly increased in group B when compared with group A (P < 0.05). RT-PCR results showed that there were significant differences in the expressions of IL-4, IL-17, and IFN-gamma mRNA between groups B, C and group A (P < 0.05); the expressions of IL-4 and IFN-gamma mRNA were significantly lower in group C than in group B (P < 0.05).
Human PMSCs transplantation can suppress the acute immunological rejection in allogeneic skin transplantation. The possible mechanism may be partially related to the inhibitory effect on the secretion of IL-17 and IFN-gamma.
探讨人胎盘间充质干细胞（PMSCs）对小鼠同种异体皮肤移植免疫排斥反应的影响。
采用自愿捐献者的胎盘胎儿组织分离培养PMSCs，实验使用第3代PMSCs。选取12日龄的30只清洁级CD1（ICR）小鼠作为同种异体皮肤移植的供体。选取68周龄的30只C57BL/6小鼠作为受体，制作直径为12 mm的背部皮肤缺损，随机分为3组（n = 10）：A组为自体移植组；B组为同种异体移植+尾静脉注射PBS组；C组为同种异体移植+尾静脉注射人PMSCs（1×10⁵个细胞/只）组。观察皮瓣存活情况。皮肤移植后7 d，分别采用ELISA法和RT-PCR法检测外周血白细胞计数、腹腔液巨噬细胞活化情况以及外周血和脾脏中白细胞介素4（IL-4）、白细胞介素17（IL-17）和干扰素γ（INF-γ）的表达水平。
A组皮瓣存活时间[（58.33±4.04）d]显著长于B组[（3.80±0.92）d]和C组[（6.80±0.82）d]（P<0.05），且C组皮瓣存活时间长于B组（P<0.05）。移植后7 d，A组外周血白细胞数为（6.32±0.45）×10⁹/L，B组为（7.45±0.52）×10⁹/L，C组为（6.35±0.39）×10⁹/L，B组外周血白细胞数显著多于A组和C组（P<0.05）。腹腔液巨噬细胞活化率A组为6.87%±2.40%，B组为7.84%±0.44%，C组为15.98%±2.87%；C组显著高于A组和B组（P<0.01）。ELISA结果显示，3组间IL-4浓度差异无统计学意义（P>0.05）。与B组比较，C组IL-17和IFN-γ浓度显著降低（P<0.05），与A组比较，B组IFN-γ浓度显著升高（P<0.05）。RT-PCR结果显示，B组、C组与A组IL-4、IL-17和IFN-γ mRNA表达差异有统计学意义（P<0.05）；C组IL-4和IFN-γ mRNA表达显著低于B组（P<0.05）。
人PMSCs移植可抑制同种异体皮肤移植中的急性免疫排斥反应。其可能机制可能与对IL-17和IFN-γ分泌的抑制作用有关。
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