Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, No. 39 Huaxiang Road, Shenyang, 110022, Liaoning, China.
Key Laboratory of Reproductive Dysfunction Diseases and Fertility Remodeling of Liaoning Province, No.39 Huaxiang Road, Shenyang, 110022, Liaoning Province, China.
Stem Cell Res Ther. 2021 Mar 12;12(1):178. doi: 10.1186/s13287-021-02255-3.
Premature ovarian insufficiency (POI) is one of the major causes of infertility. We previously demonstrated that transplantation of menstrual blood-derived stromal cells (MenSCs) effectively improved ovarian function in a murine model of POI. Recent studies indicated that mesenchymal stem cell-derived exosomes were important components in tissue repair. In this study, we investigated the therapeutic effects of MenSCs-derived exosomes (MenSCs-Exos) in a rat model of POI and its mechanism in restoring ovulation.
Ovaries of 4.5-day-old Sprague Dawley rats (SD rats) were cultured in vitro to evaluate the effects of MenSCs-Exos exposure on early follicle development. Furthermore, POI in rats was induced by intraperitoneal administration of 4-vinylcyclohexene diepoxide (VCD). Forty-eight POI rats were randomly assigned to four groups, each receiving a different treatment: PBS, MenSCs, MenSCs-Exos, and Exo-free culture supernatant of MenSCs. Estrous cyclicity, ovarian morphology, follicle dynamics, serum hormones, pregnancy outcomes, and molecular changes were investigated.
Exposure to MenSCs-Exos promoted the proliferation of granulosa cells in primordial and primary follicles in vitro and increased the expression of early follicle markers Deleted In Azoospermia Like (DAZL) and Forkhead Box L2 (FOXL2) while inhibiting follicle apoptosis. In vivo, MenSCs-Exos transplantation effectively promoted follicle development in the rat model of POI and restored the estrous cyclicity and serum sex hormone levels, followed by improving the live birth outcome. In addition, transplantation of MenSCs-Exos regulated the composition of the ovarian extracellular matrix and accelerated the recruitment of dormant follicles in the ovarian cortex and increased proliferation of granulosa cells in these follicles.
MenSCs-Exos markedly promoted follicle development in vitro and in vivo and restored fertility in POI rats, suggesting a restorative effect on ovarian functions. The therapeutic effect of MenSCs-Exos transplantation was sustainable, consistent with that of MenSCs transplantation. Our results suggested that MenSCs-Exos transplantation may be a promising cell-free bioresource in the treatment of POI.
卵巢早衰（POI）是不孕的主要原因之一。我们之前的研究表明，移植月经血源性基质细胞（MenSCs）可有效改善 POI 小鼠模型的卵巢功能。最近的研究表明，间充质干细胞衍生的外泌体是组织修复的重要组成部分。在这项研究中，我们研究了 MenSCs 衍生的外泌体（MenSCs-Exos）在 POI 大鼠模型中的治疗效果及其恢复排卵的机制。
将 4.5 天大的 Sprague Dawley 大鼠（SD 大鼠）的卵巢在体外培养，以评估 MenSCs-Exos 暴露对早期卵泡发育的影响。此外，通过腹腔内给予 4-乙烯环己烯二环氧（VCD）诱导大鼠 POI。将 48 只 POI 大鼠随机分为四组，分别接受以下不同的治疗：PBS、MenSCs、MenSCs-Exos 和 MenSCs 无外泌体培养上清液。检测动情周期、卵巢形态、卵泡动力学、血清激素、妊娠结局和分子变化。
体外实验中，MenSCs-Exos 促进了原始和初级卵泡颗粒细胞的增殖，增加了早期卵泡标志物Deleted In Azoospermia Like（DAZL）和 Forkhead Box L2（FOXL2）的表达，同时抑制了卵泡凋亡。体内实验中，MenSCs-Exos 移植可有效促进 POI 大鼠模型中的卵泡发育，恢复动情周期和血清性激素水平，随后提高活产率。此外，MenSCs-Exos 移植调节了卵巢细胞外基质的组成，加速了卵巢皮质中休眠卵泡的募集，并增加了这些卵泡中颗粒细胞的增殖。
MenSCs-Exos 显著促进了体外和体内的卵泡发育，并恢复了 POI 大鼠的生育能力，提示对卵巢功能有恢复作用。MenSCs-Exos 移植的治疗效果可持续，与 MenSCs 移植的效果一致。我们的结果表明，MenSCs-Exos 移植可能是治疗 POI 的一种有前途的无细胞生物资源。
