【摘要】 目的观察人脐带华通氏胶间充质干细胞(WJ-MSCs)移植治疗复发性自然流产小鼠的效果,并探讨其机制。方法雌性CBA/J小鼠与雄性BALB/c小鼠按1∶1比例合笼交配,制作正常妊娠小鼠模型(对照组,5只)。雌性CBA/J小鼠与雄性DBA/2小鼠按1∶1比例合笼交配,制作复发性自然流产小鼠模型20只,分为模型组、实验1组、实验2组、实验3组,每组各5只。模型组于孕1天经尾静脉注射DMEM/F12培养基0.2 mL;各实验组于孕第1天经尾静脉注射WJ-MSCs 0.2 mL(6×107个/mL),其中实验1组仅注射1次,实验2组隔天注射1次、连续2次,实验3组隔天注射1次、连续3次。分别于孕第14天处死各组小鼠,计算胚胎吸收率;取小鼠蜕膜及胎盘组织,采用Western blotting法检测IL-10、IL-17、IFN-γ蛋白,qRT-PCR法检测IL-10、IL-17、IFN-γmRNA。结果模型组及实验1组胚胎吸收率高于对照组,实验3组胚胎吸收率均低于模型组和实验1组(P均<0.05)。实验1、2组IL-17蛋白相对表达量高于对照组,实验3组低于对照组(P均<0.05)。实验1组蜕膜及胎盘组织中IFN-γmRNA相对表达量高于对照组,实验2、3组低于对照组(P均<0.05)。实验1、2、3组蜕膜及胎盘组织中IL-10 mRNA和蛋白相对表达量高于模型组,IL-17、IFN-γmRNA和蛋白相对表达量低于模型组(P均<0.05)。实验1、2、3组蜕膜及胎盘组织中的IL-10 mRNA和蛋白相对表达量递增,IL-17 mRNA和蛋白、IFN-γ蛋白相对表达量递减,除实验2、3组IFN-γmRNA表达差异无统计学意义外,各组其余指标相比,P均<0.05。结论 WJ-MSCs移植后复发性自然流产模型小鼠的胚胎吸收率降低,其机制可能与上调母-胎界面IL-10表达、下调IL-17及IFN-γ表达有关。更多还原

【Abstract】 Objective To investigate the therapeutic effect of Wharton jelly-derived mesenchymal stem cells(WJMSCs) on recurrent spontaneous abortion(RSA) and its mechanism. Methods Female CBA/J mice were separately mated with male BALB/c and DBA/2 mice to construct normal pregnant(n = 5) and RSA models(n = 20). The RSA models were randomly divided into the model group,experimental groups 1,2 and 3,five mice in each group. Mice in the model were injected with 0. 2 m L DMEM/F12 culture medium,mice in the experimental groups were injected with WJ-MSCs(6× 107/m L) by tail vein on day 1 of pregnancy,The mice in the experimental group 1 were injected once,mice in the experimental group 2 were injected every other day for 2 times,and mice in the experimental group 3 were injected every other day for 3 times. The mice were killed on day 14 of pregnancy,and we calculated the abortion rate,and their decidual and placental tissues were obtained to detect the expression of IL-10,IL-17 and IFN-γ by Western blotting and qRT-PCR,respectively. Results The embryo resorption rates in the model group and experimental group 1 were higher than that of the control group,and the embryo resorption rate in the experimental group 3 was lower than those in the model group and experimental group 1(all P < 0. 05). The protein expression of IL-17 in the experimental groups 1 and 2 was higher than that in the control group,while it was lower in the experimental group 3 than that of the control group(all P < 0. 05). The relative mRNA expression of IFN-γ in the experimental group 1 was higher than that in control group,while in the experimental groups 2 and 3,the levels were lower than that in the control group(all P < 0. 05). The relative mRNA and protein expression of IL-10 in the experimental groups 1,2 and 3 was higher,and the relative mRNA and protein expression of IL-17 and IFN-γ was lower than that in the model group(all P < 0. 05). The mRNA and protein expression of IL-10 in the decidua and placenta was gradually increased,the expression of IL-17 mRNA,protein and IFN-γ protein was gradually decreased in experiment groups 1,2 and 3. No significant difference was found in the IFN-γ mRNA expression between the experimental groups 2 and 3,but significant difference was found in the rest of the indicators(all P < 0. 05). Conclusions The embryo resorption rate in RSA mice is decreased after transplantation of WJ-MSC,which may be related to its up-regulation of IL-10 and down-regulation of IFN-γ and IL-17.更多还原