脐血间充质干细胞对薄型子宫内膜PI3K/Akt信号通路影响的实验研究

Experimental study on the effect of cord blood mesenchymal stem cells on PI3K/Akt signal pathway in thin endometrium

目的观察脐血间充质干细胞移植对大鼠薄型子宫内膜的作用,探讨其可能的作用机制。方法将30只SD大鼠随机分为假手术组、损伤组与干细胞组,每组10只。损伤组与干细胞组通过向子宫注射95%乙醇建立薄型子宫内膜模型,假手术组注射等量的生理盐水,损伤组注射乙醇后再通过宫腔注射脐血间充质干细胞悬液,损伤组与假手术组注射等量的生理盐水。观察3个动情周期后取子宫组织,分别进行H-E染色、免疫组化染色(观察血管内皮生长因子与Ki-67阳性表达)与Western blot检测(检测PI3K、Akt、p-Akt蛋白表达量)。结果损伤组子宫内膜的厚度、面积与腺体数量均明显少于正常组(P<0.05),干细胞组子宫内膜的厚度、面积与腺体数量均明显多于损伤组(P<0.05)。损伤组血管内皮生长因子与Ki-67阳性表达少于正常组(P<0.05),干细胞组血管内皮生长因子与Ki-67阳性表达多于损伤组(P<0.05)。损伤组的PI3K、p-Akt蛋白表达强度弱于正常组(P<0.05),干细胞组的PI3K、p-Akt蛋白表达强度强于损伤组,3组间的Akt蛋白表达强度无差异。结论脐血间充质干细胞移植可增加薄型子宫内膜的厚度、面积与腺体数量,促进子宫内膜细胞的增殖与血管生成,其作用机制可能与调节PI3K/Akt信号通路蛋白表达有关。

Objcetive To observe the effect of cord blood mesenchymal stem cell transplantation on the thin endometrium of rats and explore its possible mechanism.Methods Thirty SD rats were randomly divided into three groups:Sham operated group,injured group and stem cell group.The thin endometrium model of SD rats was established by injecting 95%ethanol into the uterus in the injured group and the stem cell group.The same amount of normal saline was injected into the sham operation group.The same amount of normal saline was injected into the injured group and the sham operation group.After three estrous cycles,the uterus tissues were taken for H-E staining,immunohistochemistry(VEGF and Ki-67 positive expression)and Western blot(PI3K,Akt,p-Akt protein expression).Results The thickness,area and number of glands in the injured group were significantly less than those in the normal group(P<0.05),and the thickness,area and number of glands in the stem cell group were significantly more than those in the injured group(P<0.05).The expression of VEGF and Ki-67 in the injured group was lower than that in the normal group(P<0.05),and the expression of VEGF and Ki-67 in the stem cell group was higher than that in the injured group(P<0.05).The expression intensity of PI3K and p-Akt protein in the injured group was weaker than that in the normal group(P<0.05),while that in the stem cell group was stronger than that in the injured group,and there was no difference among the three groups.Conclusion Cord blood mesenchymal stem cell transplantation can increase the thickness,area and gland number of thin endometrium,promote the proliferation and angiogenesis of endometrial cells,and its mechanism may be related to the regulation of PI3K/Akt signal pathway protein expression.