脐带间充质干细胞外泌体对大鼠卵巢功能作用及机制研究

Effect and mechanism of umbilical cord mesenchymal stem cell exosomes on ovarian function in rats

目的探讨脐带间充质干细胞外泌体(MSCs-exo)对大鼠卵巢功能的作用及机制。方法将24只大鼠随机分为空白组、模型组和干预组3组,每组各8只。空白组作为对照,模型组建立卵巢早衰大鼠模型,干预组建立卵巢早衰大鼠模型后采用MSCs-exo处理。ELISA法检测各组大鼠血清激素水平,Western blot检测卵巢组织中凋亡相关蛋白(Bax、Caspase-3)、相关通路蛋白(Ras、Erk)表达水平。提取各组大鼠卵巢颗粒细胞,模型组建立颗粒细胞损伤模型,干预组建立颗粒细胞损伤模型后采用MSCs-exo处理。CCK-8检测颗粒细胞活力,ELISA法检测颗粒细胞激素水平,流式细胞仪检测颗粒细胞凋亡情况。结果模型组、干预组干预2 d、4 d、6 d、8 d、10 d的颗粒细胞增殖率均低于空白组,且模型组低于干预组,差异有统计学意义(P<0.05)。模型组、干预组颗粒细胞雌二醇水平、颗粒细胞孕激素水平、血清雌二醇水平、血清孕激素水平均低于空白组,且模型组低于干预组,差异有统计学意义(P<0.05)。模型组、干预组颗粒细胞凋亡数量高于空白组,且模型组高于干预组,差异有统计学意义(P<0.05)。模型组、干预组大鼠卵巢组织Bax和Caspase-3表达水平高于空白组,且模型组高于干预组,差异有统计学意义(P<0.05)。模型组、干预组卵巢组织Ras、Erk蛋白表达水平低于空白组,且模型组低于干预组,差异有统计学意义(P<0.05)。结论MSCs-exo能够改善大鼠卵巢功能,其机制可能与调节Ras/Erk信号通路有关。

Objective To investigate the effect and mechanism of umbilical cord mesenchymal stem cell exosomes(MSCs-exo)on ovarian function in rats.Methods A total of 24 rats were randomly divided into blank group,model group and intervention group,with 8 rats in each group.The blank group was used as the control,the model group was established as the early ovarian failure rat model,and the intervention group was established as the early ovarian failure rat model and then treated with MSCs-exo.Serum hormone levels of each group were detected by ELISA.The expression levels of apoptosis-related proteins(Bax,Caspase-3)and related pathway proteins(Ras,Erk)in ovarian tissues were detected by Western blot.Ovarian granulosa cells were extracted from each group,the granulosa cell damage model was established in the model group,and the intervention group was treated with MSCs-exo after the granulosa cell injury model was established.CCK-8 was used to detect granulosa cell activity,ELISA was used to detect granulosa hormone level,and flow cytometry was used to detect cell apoptosis.Results The granulosa cell proliferation rates of model group and intervention group were lower than those of blank group at 2,4,6,8 and 10 days after intervention,and the model group was lower than the intervention group,the differences were statistically significant(P<0.05).The level of granule cell estradiol,granule cell progesterone,serum estradiol and serum progesterone in model group and intervention group were lower than those in blank group,and the model group were lower than those in intervention group,the differences were statistically significant(P<0.05).The number of granulosa cell apoptosis in model group and intervention group was higher than that in blank group,and the model group was higher than that in intervention group,the differences were statistically significant(P<0.05).The expression levels of Bax and Caspase-3 in ovarian tissue of model group and intervention group were higher than those of blank group,and the model group was higher than the intervention group,the difference was statistically significant(P<0.05).The expression levels of Ras and Erk protein in ovarian tissue of model group and intervention group were lower than that of blank group,and the model group was lower than that of intervention group,the differences were statistically significant(P<0.05).Conclusion The MSCs-exo can improve ovarian function in rats,and the mechanism may be related to the regulation of Ras/Erk signaling pathway.