人羊膜间充质干细胞联合GM-CSF对子宫内膜修复的机制研究

Endometrial regeneration is an important factor affecting the prognosis of endometrial injury diseases such as intrauterine adhesions, but its specific regulatory mechanism is unclear. Stem cell therapy shows potentials in promoting endometrial regeneration. Our earlier animal studies found that human amniotic mesenchymal stem cells (hAMSC) promote the repair and reconstruction of the damaged endometrial tissue, but not only the underlying mechanism of how the stem cells differentiate into endometrial cells or regulate endometrial regeneration through paracrine is unknown, neither the effect on pregnancy outcome of endometrial injured rats. Granulocyte-macrophage colony stimulating factor (GM-CSF) plays an important role in remodeling endometrium, improving the microenvironment of uterine cavity and promoting embryonic development. Besides it can promote the recruitment of stem cells to injury site and accelerates damaged tissue repair. However, it has not been fully illustrated in the literature on the aspect of endometrial regeneration. The high expression of transforming growth factor（TGF） beta 1 in the injured endometrium is closely related to the development of endometrial fibrosis.The present study observes the proliferation and apoptosis of endometrial cells in vitro, and then evaluate the endometrial tissue morphology, fibrosis, endometrial receptivity and embryo implantation rate in vivo, to further explore the potential mechanism of hAMSC and GM-CSF combination therapy, with or without TGF beta 1 on endometrial regeneration, aiming to provide a theoretical basis for clinical treatment of endometrial injury diseases.

子宫内膜修复是影响宫腔粘连等内膜损伤疾病预后的重要因素，但其具体调控机制不详。干细胞治疗在促进子宫内膜修复中的意义日益受到关注，前期动物模型研究发现hAMSC促进子宫内膜组织形态学修复的线索，但其是否通过旁分泌作用调控内膜修复尚不可知。GM-CSF在重塑子宫内膜、改善宫腔微环境、促进胚胎发育发挥重要作用，但其在子宫内膜损伤后修复的作用尚不清楚。TGF-β1在损伤子宫内膜的高表达显示其与子宫内膜纤维化的发生发展关系密切。本研究拟通过联合hAMSC与GM-CSF，观察两者通过TGF-β1在体外对子宫内膜细胞增殖、凋亡能力的影响；并进一步探讨联合作用于子宫内膜损伤大鼠模型，观察子宫内膜组织的大体形态学改变、纤维化程度、再生程度、子宫内膜容受性改变以及胚胎着床情况，探索两者调控内膜修复的潜在机制，为临床诊疗子宫内膜损伤性疾病提供理论基础。

子宫是胚胎着床、生长发育的部位，是维持女性月经生理和生育功能的重要器官。子宫内膜损伤引起的宫腔粘连和内膜纤维瘢痕化是育龄女性子宫性不孕最主要的原因，不仅使患者饱受身心摧残，也给家庭和睦与社会安定带来极大隐患。但目前临床诊疗尚没有能够改善宫腔粘连生育功能的有效治疗手段，如何促进宫腔粘连的子宫内膜修复是女性子宫生育力保护与重塑的重中之重。随着对子宫内膜再生机制的深入了解，研究发现子宫内膜干细胞与子宫内膜再生能力密切相关。从干细胞的角度理解IUA的发生机制，促使更多的研究突破传统治疗方法的局限，在干细胞水平探寻新的治疗手段。人羊膜间充质干细胞是组织工程和再生医学中广泛应用的具有多向分化能力的细胞源。. 本研究通过联合hAMSC与GM-CSF，发现其可促进TGF β1诱导的纤维化子宫内膜细胞增殖，减少细胞凋亡；且能改善子宫内膜损伤大鼠模型的内膜纤维化程度，增加内膜腺体和厚度。以上研究结果为进一步明确子宫内膜损伤性疾病发生发展机制，后期提高子宫内膜容受性，改善生育结局提供重要依据，为临床探求行之有效的子宫内膜损伤修复治疗方案具有重要的理论意义。