羊膜干细胞用于修复损伤子宫内膜的动物实验研究

Experimental Study of Amniotic Mesenchymal Stem Cells Used for Repair the Injuried Endometrium of Mouse

目的通过使用羊膜干细胞修复损伤的小鼠子宫内膜实验,了解羊膜干细胞是否有助于子宫内膜的修复,为将来进一步应用于临床提供依据。方法参照人羊膜干细胞方法先制备小鼠羊膜干细胞,再建立小鼠子宫内膜损伤模型,然后根据方法不同分为3组(造模组50只,假手术组20只,对照组20只)。造模术后10 d取造模组10只,假手术组10只,对照组10只处死,肉眼观察宫腔改变,取出各组小鼠子宫置入中性甲醛固定,HE及Masson染色,对其进行纤维化半定量评分。各组再取10只小鼠与雄性小鼠合笼饲养,妊娠10 d开腹记数着床胚胎数作为评价生育能力指标。另外30只造模组行粘连分离后分别给予宫腔内注射羊膜干细胞、口服雌激素及空白。结果模型组在造模后宫腔纤维化面积比例增加,腺体数量减少,妊娠能力明显下降,而假手术组及对照组无明显变化,造模组与假手术组及对照组相比,3项指标都有显著性差异(P<0.05),对造模成功组采用不同治疗方案,结果显示,干细胞宫腔内注射可以改善宫腔纤维化面积及腺体数量,而雌激素无明显作用,3组治疗相比,干细胞组比其他组作用更明显,差异有显著性((P<0.05)。结论宫腔粘连分离后注入羊膜干细胞有利于子宫内膜的修复。

Objective To investigate whether the amniotic mesenchymal stem cells（ m AMC） could repair injured mouse endometrium. Methods Female moue amniotic mesenchymal stem cells（ m AMC） were established with reference to the method that used in human Amniotic mesenchymal stem cells collection. The mouse models of the injured mouse endometrium were established and then were divied into three groups（ model qroup,n = 50,false group,n = 20 and normal control group,n = 20） according to the methods used,10 days after operation. 10 mice were selected in every group and were killed and change of uterine cavity were observed,then uterines were fixed with neutral formalin and then were dyed with HE and Masson. The fibrosis of endometrium was evaluated. another 10 female mice in each group were reared with male mice,the number of implanted embryo was counted as indication of fertility after 10 days of pregnancy. Another 30 mice in model group were treated with m AMC,estrogen and empty after separation of fibre adhesion and the reparation of injured endometrium after transplanted stem cells were observed. Results After opertion,the fibrosis of endometrium increased,the number of gland decreased and the fertility droped in model group,there was no change in other groups. There was a obvious difference between operation group and other group（ P〈0. 05）. In the model group,different treatment were used after separation of fibre adhesion,m AMC intrauterin- injected could obviously improved the result of repairation. Conclusion m AMC intrauterin- injected could obviously improved the result of repairation after separation of fibre adhesion in uterine.