生殖医学杂志

目的 探讨人脐带间充质干细胞(hUCMSCs)尾静脉注射对同种免疫型复发性流产(RSA)模型小鼠治疗的有效性。方法 CBA/J雌鼠与DBA/2雄鼠交配建立同种免疫型RSA模型，按照随机数字表法进行随机分组，一组给予hUCMSCs治疗，为hUCMSCs治疗组(n=8);另一组不进行hUCMSCs治疗，为RSA模型组(n=8)。CBA/J雌鼠与BALB/c雄鼠交配建立正常妊娠模型(正常妊娠组，n=8)。hUCMSCs治疗组尾静脉注射hUCMSCs悬液，正常妊娠组和RSA模型组注射等量生理盐水。妊娠第14天麻醉小鼠，称重并取材，暴露子宫，计算胚胎吸收率。采用流式细胞术检测脾脏组织免疫细胞标志物，苏木精-伊红(HE)染色胎盘组织，酶联免疫吸附试验(ELISA)检测血清中炎症因子的表达水平。结果 胎盘组织HE染色结果显示，相较正常妊娠组，RSA模型组呈现明显病理改变，经hUCMSCs治疗后，hUCMSCs治疗组细胞形态结构、胞浆基质等基本恢复至正常妊娠组胎盘组织形态。正常妊娠组和hUCMSCs治疗组小鼠的存活胚胎数显著高于RSA模型组，胚胎吸收率显著低于RSA模型组(P均<0.01),脾脏组织内Th1/Th2细胞比值、Th17/Treg细胞比值以及NK细胞占比均显著低于RSA模型组(P<0.01),血清中的促炎因子干扰素(IFN)-γ和白介素(IL)-17A水平均显著低于RSA模型组(P<0.01),但抗炎因子IL-4与IL-10水平均显著高于RSA模型组(P<0.01)。结论 hUCMSCs尾静脉注射能有效改善同种免疫型RSA小鼠的妊娠结局，其治疗效应可能与恢复Th1/Th2及Th17/Treg细胞轴平衡有关。

Objectives:To investigate the efficacy of human umbilical cord mesenchymal stem cells(hUCMSCs) administered via tail vein injection for the treatment of model mice with a homozygous immune recurrent miscarriage(RSA).Methods:The homozygous immune RSA model was established by mating CBA/J female mice with DBA/2 male mice. According to the random number table, mice were randomly divided into two groups. One group was treated with hUCMSCs, namely hUCMSCs treatment group(n=8). The other group did not receive treatment with hUCMSCs as the RSA model group(n=8). A normal pregnancy model was established by mating CBA/J female mice with BALB/c male mice(n=8). The hUCMSCs treatment group underwent tail vein injection of hUCMSCs suspension, while the normal pregnancy group and RSA model group received equal amounts of physiological saline injection. On the 14^th day of pregnancy, the anesthetized mice were weighed and samples were taken, and the uterus was exposed and the embryo absorption rate was evaluated. Flow cytometry was used to detect immune cell markers in spleen tissue, hematoxylin eosin(HE) staining was performed on placental tissue, and enzyme-linked immunosorbent assay(ELISA) was used to detect the expression levels of inflammatory factors in serum.Results:The HE staining results of placental tissue showed that compared with the normal pregnancy group, the RSA model group exhibited significant pathological changes. After treatment with hUCMSCs, the cell morphology and cytoplasmic matrix of the hUCMSCs treatment group were basically recovered to the normal placental tissue morphology of the pregnancy group. In the normal pregnancy group and hUCMSCs treatment group, the average number of surviving embryos was significantly higher than that in the RSA model group(P<0.01),the average embryo absorption rates were significantly lower than that in the RSA model group(P<0.01),the Th1/Th2 ratio, Th17/Treg ratio and the proportion of NK cells in the spleen tissue were significantly lower than those in the RSA model group(P<0.01),the levels of pro-inflammatory cytokines of interferon(IFN)-γ and interleukin(IL)-17A in the serum were significantly lower than those in the RSA model group(P<0.01),but the levels of anti-inflammatory cytokines in terms of IL-4 and IL-10 were significantly higher than those in the RSA model group(P<0.01).Conclusions:The tail vein injection of hUCMSCs can effectively improve the pregnancy outcomes of homologous immune RSA mice, and its therapeutic effect may be related to restoring the balance of Th1/Th2 and Th17/Treg cell axis.