Department of obstetrics and gynecology, Qilu Hospital, Shandong University, 107 Wenhua Xi Road, Jinan, 250012, Shandong Province, P.R. China.
Cryomedicine Laboratory, Qilu Hospital, Shandong University, 107 Wenhua Xi Road, Jinan, 250012, Shandong Province, P.R. China.
Sci Rep. 2017 May 31;7(1):2552. doi: 10.1038/s41598-017-02786-x.
Human umbilical cord mesenchymal stem cells (huMSCs) can treat primary ovarian insufficiency (POI) related to ovarian granulosa cell (OGC) apoptosis caused by cisplatin chemotherapy. Exosomes are a class of membranous vesicles with diameters of 30-200 nm that are constitutively released by eukaryotic cells. Exosomes mediate local cell-to-cell communication by transferring microRNAs and proteins. In the present study, we demonstrated the effects of exosomes derived from huMSCs (huMSC-EXOs) on a cisplatin-induced OGC model in vitro and discussed the preliminary mechanisms involved in these effects. We successfully extracted huMSC-EXOs from huMSC culture supernatant and observed the effective uptake of exosomes by cells with fluorescent staining. Using flow cytometry (with annexin-V/PI labelling), we found that huMSC-EXOs increased the number of living cells. Western blotting showed that the expression of Bcl-2 and caspase-3 were upregulated, whilst the expression of Bax, cleaved caspase-3 and cleaved PARP were downregulated to protect OGCs. These results suggest that huMSC-EXOs can be used to prevent and treat chemotherapy-induced OGC apoptosis in vitro. Therefore, this work provides insight and further evidence of stem cell function and indicates that huMSC-EXOs protect OGCs from cisplatin-induced injury in vitro.
人脐带间充质干细胞(huMSCs)可治疗顺铂化疗引起的卵巢颗粒细胞(OGC)凋亡导致的原发性卵巢功能不全(POI)。外泌体是一类直径为 30-200nm 的膜性囊泡,由真核细胞组成性释放。外泌体通过转移 microRNAs 和蛋白质来介导细胞间的局部通讯。在本研究中,我们证明了 huMSCs 来源的外泌体(huMSC-EXOs)对体外顺铂诱导的 OGC 模型的作用,并讨论了这些作用涉及的初步机制。我们成功地从 huMSC 培养上清液中提取了 huMSC-EXOs,并通过荧光染色观察到细胞对 exosomes 的有效摄取。使用流式细胞术(annexin-V/PI 标记),我们发现 huMSC-EXOs 增加了活细胞的数量。Western blot 显示,Bcl-2 和 caspase-3 的表达上调,而 Bax、cleaved caspase-3 和 cleaved PARP 的表达下调,从而保护 OGC。这些结果表明,huMSC-EXOs 可用于预防和治疗体外化疗诱导的 OGC 凋亡。因此,这项工作为干细胞功能提供了深入的了解和进一步的证据,并表明 huMSC-EXOs 可保护 OGC 免受顺铂诱导的损伤。