Reproductive Medicine Center, The 901st Hospital, Hefei, China; Department of Obstetrics and Gynecology, Maternal and Child Health Hospital, Anhui Province, Hefei, China.
Reproductive Medicine Center, The 901st Hospital, Hefei, China.
Taiwan J Obstet Gynecol. 2020 Jul;59(4):527-533. doi: 10.1016/j.tjog.2020.05.010.
Long term exposure to gonadotoxic chemotherapy is becoming a major cause of premature ovarian failure/insufficiency (POF/POI) with the increasing cancer incidence among young women. The present study was designed to investigate the protective effects of human cord mesenchymal stem cells (HUCMSCs)-derived extracellular vesicles (EVs) on cisplatin (CDDP)-damaged granulosa cells (GCs) in vitro.
EVs were obtained from supernatant of cultured HUCMSCs by ultracentrifugation method, purified by Sucrose density gradient centrifugation, and then were co-cultured with cisplatin-damaged GCs of 3-weeks female Sprague-Dawley (SD) rats. PKH26 labeled EVs could be observed in normal and CDDP-damaged GCs after 6 h co-culture.
The surviving GCs were significantly higher and apoptotic GCs were significantly lower in EVs + CDDP group compared with CDDP group. Meanwhile, the levels of E2 and StAR (the key gene related to synthesis of steroid hormone) were significantly higher in EVs + CDDP group compared with CDDP group. Furthermore, the mRNA expression of Caspase 3 was down-regulated significantly and the ratio of Bcl-2/Bax was up-regulated significantly in EVs + CDDP group. Moreover, the protective effect of EVs on CDDP-damaged GCs showed a dose-dependent effect.
HUCMSCs-derived EVs could become incorporated to CDDP-damaged GCs, and increase the number of living cells, therefore playing important roles in promoting resistance to cisplatin-induced GCs apoptosis and restoring synthesis and secretion of steroid hormone in GCs. This study might provide a theoretical and experimental basis for use of mesenchymal stem cells (MSCs) derived EVs instead of MSCs as a cell-free therapeutic strategy for the patients with POI induced by chemotherapeutic agents.
长期暴露于性腺毒性化疗药物是导致年轻女性卵巢早衰/功能不全（POF/POI）的主要原因，随着癌症发病率的增加，这种情况越来越常见。本研究旨在探讨人脐带间充质干细胞（HUCMSCs）衍生的细胞外囊泡（EVs）对顺铂（CDDP）损伤的颗粒细胞（GCs）的保护作用。
超速离心法从培养的 HUCMSC 上清液中提取 EVs，通过蔗糖密度梯度离心法纯化，然后与 3 周龄雌性 Sprague-Dawley（SD）大鼠的顺铂损伤的 GCs 共培养。共培养 6 h 后，PKH26 标记的 EVs 可在正常和 CDDP 损伤的 GCs 中观察到。
与 CDDP 组相比，EVs+CDDP 组存活的 GCs 明显增多，凋亡的 GCs 明显减少。同时，EVs+CDDP 组 E2 和 StAR（与类固醇激素合成相关的关键基因）水平明显高于 CDDP 组。此外，EVs+CDDP 组 Caspase 3 的 mRNA 表达明显下调，Bcl-2/Bax 比值明显上调。此外，EVs 对 CDDP 损伤的 GCs 的保护作用呈剂量依赖性。
HUCMSC 衍生的 EVs 可被整合到 CDDP 损伤的 GCs 中，增加活细胞数量，因此在促进顺铂诱导的 GCs 凋亡抵抗和恢复 GCs 类固醇激素合成和分泌方面发挥重要作用。本研究为使用间充质干细胞（MSCs）衍生的 EVs 替代 MSCs 作为化疗药物诱导的 POI 患者的无细胞治疗策略提供了理论和实验依据。
