人羊膜上皮细胞移植对子宫瘢痕模型大鼠子宫内膜的影响及机制

Effect and mechanism of transplantation of human amniotic epithelial cells on endometrium of uterine scar model rats

目的 探讨人羊膜上皮细胞(hAECs)移植对大鼠子宫瘢痕模型子宫内膜的改善及对基质金属蛋白酶8(MMP-8)、血管内皮生长因子(VEGF)的影响。方法 建立大鼠子宫瘢痕模型,并随机分为模型组及移植组,每组各18只,另取18只大鼠作为假手术组。移植组大鼠于子宫瘢痕处注射hAECs,模型组及假手术组仅给予等量PBS。4周后,各组取8只大鼠子宫组织,HE染色和Masson染色分别观察组织形态学变化和纤维化情况,测量子宫内膜厚度和腺体数量;细胞角蛋白和整合素β3免疫组化染色分别评估子宫内膜生长和容受性;RT-qPCR技术检测子宫内膜组织中MMP-8、VEGFA mRNA表达水平;Western blot法检测组织中MMP-8、VEGFA蛋白表达水平;8周后,取各组剩余10只大鼠进行妊娠能力测定。结果 模型组及移植组大鼠子宫内膜厚度、腺体数、角蛋白和整合素β3 IOD值、MMP-8、VEGFA mRNA和蛋白相对表达水平、妊娠率和子宫胚胎数低于假手术组(P<0.05);移植组大鼠子宫内膜厚度、腺体数、角蛋白和整合素β3 IOD值、MMP-8、VEGFA mRNA和蛋白相对表达水平、妊娠率和子宫胚胎数高于模型组(P<0.05),另外,hAECs移植可改善子宫瘢痕大鼠子宫内膜组织病理形态,减轻子宫内膜纤维化程度。结论 hAECs移植可改善子宫内膜损伤,减少瘢痕形成,提高子宫内膜容受性,并增强模型大鼠的妊娠功能,其作用机制可能与促进MMP-8和VEGFA表达有关。

Objective To investigate the effect of human amniotic epithelial cell(hAEC)transplantation on endometrium improvement and matrix metalloproteinase 8(MMP-8)and vascular endothelial growth factor(VEGF)expression in a rat model of uterine scaring.Methods The uterine scar model was established in rats that were randomly divided into model and transplantation groups with 18 rats in each group.The other 18 rats were used as the sham operation group.Rats in the transplantation group were injected with hAECs in the uterine scar,and rats in model and sham operation groups were administered the same amount of PBS.After 4 weeks,the uterine tissues of eight rats in each group were collected.Histomorphological changes and endometria fibrosis were observed by HE staining and Masson staining respectively,and the endometrial thickness and number of glands were measured.Endometrial growth and receptivity were evaluated by immunohistochemical staining of cytokeratin and integrinβ3,respectively.mRNA expression of MMP-8 and VEGFA in endometrial tissues was measured by RT-qPCR.Western blot was used to measure MMP-8 and VEGFA protein expression.After 8 weeks,the remaining 10 rats in each group were used to assess gestational ability.Results The endometrial thickness,gland number,IOD value of keratin and integrinβ3,relative mRNA and protein expression levels of MMP-8 and VEGFA,pregnancy rate and number of uterine embryos in model and transplantation groups were lower than those in sham operation group(P<0.05).The endometrial thickness,gland number,IOD value of keratin and integrinβ3,relative mRNA and protein expression of MMP-8 and VEGFA,pregnancy rate and number of uterine embryos were higher than those in model group(P<0.05).Additionally,hAEC transplantation improved the pathological morphology of endometrial tissue in rats with uterine scaring and reduced the degree of endometrial fibrosis.Conclusions hAEC transplantation improves endometrial injury,reduces scar formation,improves endometrial receptivity,and enhances pregnancy function in model rats,which may be related to promotion of MMP-8 and VEGFA expression.