人早孕蜕膜间充质干细胞移植治疗大鼠宫腔粘连

目的:探讨人早孕蜕膜间充质干细胞(hDMSCs)治疗大鼠宫腔粘连(IUA)的可行性和有效性。方法:体外分离、培养、纯化、鉴定h DMSCs。将30只雌性SD大鼠随机分为3组:蜕膜间充质干细胞治疗组(IUA+hDMSCs)、IUA模型组和假手术(sham)组,每组各10只。IUA+hDMSCs组和IUA组复制IUA模型后10 min内,IUA+hDMSCs组经尾静脉注射2×10~6个Dio标记的hDMSCs,每3天补充1次,连续注射3次;IUA组经尾静脉注射相同容量的PBS;sham组仅行开腹手术。3个动情周期后取子宫行HE染色和Masson染色,测量子宫内膜厚度、子宫内膜腺体数目、子宫内膜小血管数目和纤维化面积比率,并行免疫组化检测子宫内膜组织细胞角蛋白、波形蛋白、转化生长因子(TGF)-β1及整合素αγβ3的表达量,q PCR检测IUA相关基因含量,免疫荧光观察h DMSCs在内膜中的分布情况,并评估大鼠的生育能力。结果:IUA组部分子宫腔闭锁,内膜萎缩,腺体减少,大量纤维化改变。IUA+hDMSCs组与IUA组相比子宫间质厚度明显增加(P0.05),大量腺体增生和小血管新生(P0.05),纤维化程度减小(P0.05),与sham组比较无统计学差异。IUA+hDMSCs组细胞角蛋白、波形蛋白和整合素αγβ3表达量明显高于IUA组(P0.05),而TGF-β1表达量低于IUA组(P0.05)。qPCR结果显示与IUA组相比,除TNF-α和结缔组织生长因子下调外,IUA+hDMSCs组的干细胞因子、血管内皮生长因子、碱性成纤维细胞生长因子和thrombospondin-1均明显上调。免疫荧光结果显示Dio- DMSCs主要分布于腔上皮层,部分位于间质。IUA+hDMSCs组大鼠双侧子宫均妊娠,平均胎鼠数量与IUA组相比有统计学差异(P0.05)。结论:hDMSCs可在受损的大鼠子宫内膜中存活、增殖,有效修复大鼠的子宫内膜,抑制IUA。

Therapeutic effect of first-trimester human decidual mesenchymal stem cell transplantation on intrauterine adhesions in rats

AIM: To investigate the feasibility and therapeutic effect of transplantation with first-trimester human decidual mesenchymal stem cells (hDMSCs) on intrauterine adhesions (IUA) in rats. METHODS: The hDMSCs were isolated, cultured, purified and identified in vitro. Female SD rats were randomly divided into IUA+hDMSCs group, IUA group and sham group. The rats in IUA group were injected with 1 mL PBS via the tail vein, while the rats in IUA+hDMSCs group were injected with 2×10⁶ hDMSCs with 3 consecutive injections at every 3-day intervals. The rats in sham group only received laparotomy. The general, histological, and immunohistochemical observations were performed 3 estrous cycles postoperatively. The expression of IUA-related genes was analyzed by quantitative PCR. Immunofluorescence and pregnancy test were also performed. RESULTS: The modeled uteri were of partial atresia and fibrosis, the endometrium was atrophic and the numbers of gland decreased. The endometrium was significantly thicker, the numbers of gland and small vessels increased, and the fibrosis decreased in IUA+hDMSCs group compared with IUA group (P < 0.05). The expression of cytokeratin, vimentin and integrin αγβ3 was significantly stronger in IUA+hDMSCs group than that in IUA group, whereas the expression of TGF-β1 was weaker than that in IUA group (P < 0.05). The average number of litters in right horns of IUA+hDMSCs group was more than that in IUA group (P < 0.05). Except for down-regulation of TNF-α and CTGF, compared with IUA group, the mRNA expression levels of SCF, VEGF, bFGF, IGF-1 and TSP-1 were increased in IUA+hDMSCs group. Dio-labeled hDMSCs mainly located in endometrial epithelium.CONCLUSION: The first-trimester human decidual mesenchymal stem cells survive and proliferate in the rat endometrium and regenerate the endometrium. hDMSC transplantation is a potential novel treatment for IUA and may also be useful to prevent IUA after uterine injury.