Exosomal miRNA-17-5p derived from human umbilical cord mesenchymal stem cells improves ovarian function in premature ovarian insufficiency by regulating SIRT7
Exosomal miRNA-17-5p derived from human umbilical cord mesenchymal stem cells improves ovarian function in premature ovarian insufficiency by regulating SIRT7
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源自人脐带间充质干细胞的外泌体 miRNA-17-5p 通过调节 SIRT7 改善卵巢早衰患者的卵巢功能
DOI：
10.1002/stem.3204
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发表时间：
2020-05-29
期刊：
影响因子：
5.2
通讯作者：
Huang, Boxian
中科院分区：
文献类型：
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作者：
Ding, Chenyue;Zhu, Liping;Huang, Boxian
卵巢功能不全(POI)在临床上对40岁以上的女性是不可逆转的。虽然许多研究表明间充质干细胞治疗效果令人满意，但其潜在的治疗机制仍不清楚。从人脐带间充质干细胞(HUMSCs)的培养液中收集外体，用电子显微镜和蛋白质印迹(WB)分析进行鉴定。然后，在环磷酰胺(CTX)损伤的人颗粒细胞(HGCs)的培养液中加入外切体，并将其注射到CTX诱导的POI模型小鼠的卵巢中，检测抗凋亡和凋亡基因的表达。接下来，通过小RNA测序检测hUMSC来源的外切体(hUMSC-Exos)的microRNA表达谱。在评价卵巢表型和功能、ROS水平和SIRT7表达之前，通过miR-17-5p敲除miR-17-5p证实外体microRNA-17-5p(miR-17-5p)的改善作用。最后，通过RNA干扰或逆转录病毒转导抑制或过表达SIRT7，并分析PARP1、Gamma H 2AX和XRCC6的蛋白表达。验证了hUMSC-Exos对POI的改善作用。我们的结果表明，hUMSC-Exos可恢复POI小鼠模型的卵巢表型和功能，促进CTX损伤的hGCs和卵巢细胞的增殖，并通过传递外体miR-17-5p和抑制SIRT7的表达来减轻ROS的聚集。此外，我们的研究结果表明，miR-17-5p通过抑制SIRT7抑制PARP1、Gamma H2AX和XRCC6。我们的发现提示外体miR-17-5p及其下游靶基因SIRT7在hUMSC移植治疗中具有重要作用。这项研究表明，基于外切体的治疗POI是有希望的。
Premature ovarian insufficiency (POI) is clinically irreversible in women aged over 40 years. Although numerous studies have demonstrated satisfactory outcomes of mesenchymal stem cell therapy, the underlying therapeutic mechanism remains unclear. Exosomes were collected from the culture medium of human umbilical cord mesenchymal stem cells (hUMSCs) and assessed by electron microscopy and Western blot (WB) analysis. Then, exosomes were added to the culture medium of cyclophosphamide (CTX)-damaged human granulosa cells (hGCs), and the mixture was injected into the ovaries of CTX-induced POI model mice before detection of antiapoptotic and apoptotic gene expression. Next, the microRNA expression profiles of hUMSC-derived exosomes (hUMSC-Exos) were detected by small RNA sequencing. The ameliorative effect of exosomal microRNA-17-5P (miR-17-5P) was demonstrated by miR-17-5P knockdown before assessment of ovarian phenotype and function, reactive oxygen species (ROS) levels and SIRT7 expression. Finally, SIRT7 was inhibited or overexpressed by RNA interference or retrovirus transduction, and the protein expression of PARP1, gamma H2AX, and XRCC6 was analyzed. The ameliorative effect of hUMSC-Exos on POI was validated. Our results illustrated that hUMSC-Exos restored ovarian phenotype and function in a POI mouse model, promoted proliferation of CTX-damaged hGCs and ovarian cells, and alleviated ROS accumulation by delivering exosomal miR-17-5P and inhibiting SIRT7 expression. Moreover, our findings elucidated that miR-17-5P repressed PARP1, gamma H2AX, and XRCC6 by inhibiting SIRT7. Our findings suggest a critical role for exosomal miR-17-5P and its downstream target mRNA SIRT7 in hUMSC transplantation therapy. This study indicates the promise of exosome-based therapy for POI treatment.