人脐带间充质干细胞移植对大鼠流产模型妊娠结局的影响

Effect of transplantation of Wharton jelly-derived mesenchymal stem cells on pregnancy outcome in rats of abortion model

目的探讨人脐带间充质干细胞(WJ-MSCs)移植对大鼠流产模型妊娠结局的影响及机制。方法将48只孕鼠随机分为6组,每组8只。生理盐水对照组(a):孕6~8d皮下注射生理盐水0.4mg·kg^(-1)·d^(-1),孕9d尾静脉注射生理盐水1ml/kg;乙醇对照组(b):孕6~8d皮下注射75%乙醇0.4mg·kg^(-1)·d^(-1),孕9d尾静脉注射生理盐水1ml/kg;流产模型组(c):孕6~8d皮下注射溴隐亭0.4mg·kg^(-1)·d^(-1),孕9d尾静脉注射生理盐水1ml/kg;实验组d1、d2、d3分别于孕6~8d皮下注射溴隐亭0.4mg·kg^(-1)·d^(-1),孕9d尾静脉注射WJ-MSCs 1×105个、1×106个、1×107个(生理盐水使用量为1ml/kg,将细胞制成混悬液后注射)。各组大鼠均于孕14d处死并取材,观察各组大鼠胚胎吸收率,RT-PCR检测蜕膜组织白介素-10(IL-10)、干扰素-γ(IFN-γ)和IL-17mRNA的相对表达量。ELISA检测各组外周血IL-10、IFN-γ和IL-17的含量。结果 c组和d1组的平均胚胎吸收数和平均胚胎吸收率均显著高于a、b、d2、d3组(P<0.05)。c组、d1组与a、b、d2、d3组比较,IL-10mRNA表达显著降低(P<0.05),IFN-γ和IL-17mRNA表达显著升高(P<0.05)。c组、d1组与a、b、d2、d3组比较外周血IL-10含量显著降低(P<0.05),IFN-γ和IL-17含量显著升高(P<0.05)。c组与d1组间,a、b组与d2、d3组间的观察指标比较均无显著性差异(P>0.05)。结论一定浓度的WJ-MSCs移植入大鼠流产模型体内,可以降低胚胎吸收率,改善大鼠流产模型的妊娠结局;其机制可能与WJ-MSCs上调IL-10、下调IFN-γ和IL-17的含量有关。

Objective： To investigate the effect of transplantation of Wharton jelly-derived mesenchymal stem cells （WJ-MSCs） on the pregnancy outcome in the rats of abortion model. Methods： Forty eight pregnancy rats were divided into six groups and eight for each group： the rats were given subcutaneous injection of saline （0.4 mg·kg^-1·d^-1） from the 6^th to 8^th of pregnancy,and tail intravenous injection of saline （1 ml/kg） on the 9^th day of pregnancy in saline control group （group a） ;the rats were given subcutaneous injection of 75% ethanol （0.4 mg·kg^-1·d^-1） from the 6^th to 8^th of pregnancy,and tail intrayenous injection of saline （1 ml/kg） on the 9th day in ethanol control group （group b） ;the rats were given subcutaneous injection of bromocriptine （0.4 mg·kg^-1·d^-1） from the 6^th to 8^th of pregnancy,and tail intravenous injection of saline （1 ml/kg） on the 9th day in miscarriage model group （group c）;the rats given subcutaneous injection of bromoeriptine （0.4 mg·kg^-1·d^-1） from the 6th to 8th of pregnancy, and tail intravenous injection of 1 ×10^5, 1 ×10^6 and 1 ×10^7 WJ-MSCs respectively in group d1,d2 and d3 respectively. All the rats were killed and collected the tissue materials at the 14^th gestation days. The embryo resorption rate was observed in every group. The mRNA expression of interleukin （IL）-10,interferon-γ （IFN-γ） and IL-17 in the decidual tissues were detected by RT-PCR. The levels of IL-10,IFN-γ and IL-17 in peripheral blood were detected by ELISA. Results： Embryos absorption rate in group c and group dl was significantly higher than that in group a,b, d2 ＆ d3 （P〈 0.05）. The mRNA expression of IL-10 was significantly decreased, but mRNA expressions of IFN-γ IL-17 were significantly increased in group C ＆ d1 compared with group a,b,d2 ＆ d3 （P〈0.05）. The IL-10 level was significantly decreased and IFN-γ ＆ IL-17 levels were significantly increased in peripheral blood in group c ＆ d1 compared with group a,b,d2 ＆ d3 （P〈0.05）. There were no significant differences in the observation index between group c and dl, group a, b and group d2, d3 （P〉0.05）. Conclusions： Transplanting certain amount of WJ-MSCs into the rats of abortion model may reduce the embryo absorptivity,and improve the outcome of pregnancy. The mechanism maybe related with WJ- MSCs up-regulating the expression of IL-10 and down-regulating the expression of IFN-γ and IL-17.