摘要
目的 评估人脐带间充质干细胞外泌体(hUCMSC-Exos)对米非司酮诱导的子宫内膜基质细胞(hESCs)损伤的逆转作用,并筛选hUCMSC-Exos中发挥作用的microRNA.方法 以米非司酮(mifepristone)、人脐带间充质干细胞来源外泌体(hHUCMSC-Exos)和miRNA单独或联合作用于子宫内膜基质细胞(hESCs),分为mifepristone组、mifepristone+hHUCMSC-Exos组、mifepristone+miRNA组,加入等量生理盐水的hESCs为对照组.分组干预处理后,MTT法检测hESCs存活率变化;流式细胞术检测hESCs凋亡率变化;实时荧光定量PCR实验检测hESCs中肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)mRNA表达水平的影响.结果 与对照组比较,随着米非司酮浓度的增加以及作用时间的延长,hESCs存活率下降(P＜0.01).与mifepristone组比较,加入hUCMSC-Exos可以提高hESCs的存活率、降低hESCs的凋亡率,减少hESCs炎性因子TNF-α、IL-6、IL-iβ mRNA表达(P＜0.01).与 mifepristone 组比较,hESCs 转染 hUCMSC-Exos 中相对表达量较高的 10 种 microRNA(miR-143-3p、miR-181-5p、miR-127-3p、miR-423-3p、miR-222-3p、miR-7i-5p、miR-22-3p、miR-221-3p、miR-100-5p、miR-21-5p)后,miR-143-3p可以提高hESCs的存活率(P＜0.01).与mifepristone组比较,转染miR-143-3p降低hESCs的凋亡率、减少hESCs 炎性因子 TNF-α、IL-6、IL-1β mRNA 表达(P＜0.01).结论 hUCMSC-Exos 通过 miR-143-3p 提高米非司酮损伤的子宫内膜基质细胞存活,降低凋亡发生以及炎性因子的释放.
Abstract
Objective To evaluate the reversal effect of human umbilical cord mesenchymal stem cell exosomes(hUCMSC-Exos)on mifepriston-induced endometrial stromal cells(hESCs)injury,and to screen the micrornas involved in the regulation of hUCMSC-Exos.Methods Endometrial stromal cells(hESCs)were treated with mifepristone,hu-man umbilical cord mesenchymal stem cell-derived exosomes(hHUCMSC-Exos)and miRNA alone or in combination.hESCs were divided into mifepristone group,mifepristone+hHUCMSC-Exos group and mifepristone+miRNA group,and hESCs added with the same amount of normal saline were used as control group.After intervention,the viability of hESCs was detected by MTT assay.Flow cytometry was used to detect the apoptosis rate of hESCs.The expression levels of tumor necrosis factor(TNF-a),interleukin-6(IL-6)and interleukin-iβ(IL-iβ)mRNA in hESCs were detected by real-time fluorescence quantitative PCR.Results Compared with the control group,the survival rate of hESCs de-creased with the increase of mifepristone concentration and the extension of mifepristone treatment time(P<0.01).Compared with the mifepristone group,the addition of hUCMSC-Exos could improve the survival rate of hESCs,reduce the apoptosis rate of hESCs,and reduce the expression of inflammatory factors TNF-α,IL-6,IL-1β mRNA(P<0.01).Compared with the mifepristone group,Ten micrornas(miR-143-3p,miR-181-5p,miR-127-3p,miR-423-3p,miR-222-3p,miR-7i-5p,Mir-22-3p and miR-22)with high relative expression in hESCs transfected hUCMSC-Exos 1-3p,miR-100-5p,miR-21-5p),miR-143-3p could improve hESCs survival rate(P<0.01).Compared with mifepristone group,transfection of miR-143-3 p reduced the apoptosis rate of hESCs and the expression of inflammatory factors TNF-α,IL-6,IL-1β mRNA(P&4t;0.01).Conclusion hUCMSC-Exos can improve the survival of mifepristone-damaged endometrial stromal cells and reduce the apoptosis and the release of inflammatory factors through miR-143-3p.
基金项目
吉林省科技发展计划(YDZJ202102CXJD076)
吉林省科技发展计划(20230203051SF)
吉林省科技发展计划(20230204036YY)
吉林省科技发展计划(20240602029RC)
吉林省卫生健康科技能力提升项目(20211c059)
NETL
NSTL
万方数据