Center of Reproductive Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China.
Center of Reproductive Medicine, The Guangxi Zhuang Autonomous Region Family Planning Research Center, Nanning, Guangxi, China.
Biol Reprod. 2019 Jul 1;101(1):91-101. doi: 10.1093/biolre/ioz077.
Peripheral blood mononuclear cells (PBMCs) are rich in hematopoietic cells and mesenchymal stem cells. Platelet-rich plasma (PRP) is rich in various growth factors. PBMCs and PRP have been suggested, individually, to restore ovarian function by improving the local microenvironment. The current study investigated the effect of granulocyte colony-stimulating factor (G-CSF)-mobilized PBMCs combined with PRP on restoring ovarian function in rats with primary ovarian insufficiency (POI). Thirty adult female rats were randomly subdivided into five groups: normal control (control), cyclophosphamide (CTX) plus subsequent PBS (POI + PBS), CTX plus subsequent PRP (POI + PRP), CTX plus subsequent G-CSF-mobilized PBMCs (POI + PBMCs), and CTX plus subsequent G-CSF-mobilized PBMCs combined with PRP (POI + PBMCs + PRP). CTX exposure induced the typical POI phenotype with increased diestrus; shortened estrus; follicle arrest at all stages; decreased serum levels of estradiol-17β (E2) and anti-Mullerian hormone (AMH); and increased levels of follicle-stimulating hormone (FSH). Transplantation of mobilized PBMCs with PRP resulted in a much earlier restoration of the estrous cycle, sex hormone levels, and preantral follicle growth in POI rats. Expression of the male-specific Sry gene in the ovarian tissues of POI + PBMCs + PRP female recipient rats was evident at 5, 10, and 20 days posttransplantation along with significant increases in the expression of angiogenesis markers CD34+ and VEGF and folliculogenesis markers AMH and FSHR. Additionally, PBMCs in combination with PRP mitigated granulosa cell apoptosis by downregulating BAX and upregulating BCL-2. These results demonstrate that G-CSF-mobilized PBMCs combined with PRP accelerate the restoration of ovarian function in POI rats by increasing ovarian neovascularization, reducing granulosa cell apoptosis, and promoting folliculogenesis.
外周血单个核细胞（PBMCs）富含造血细胞和间充质干细胞。富含各种生长因子的富血小板血浆（PRP）。单独使用 PBMCs 和 PRP 已被证明可以通过改善局部微环境来恢复卵巢功能。本研究探讨了粒细胞集落刺激因子（G-CSF）动员的 PBMCs 与 PRP 联合应用对原发性卵巢功能不全（POI）大鼠卵巢功能恢复的影响。30 只成年雌性大鼠被随机分为五组：正常对照组（对照组）、环磷酰胺（CTX）加随后的磷酸盐缓冲液（POI+PBS）、CTX 加随后的 PRP（POI+PRP）、CTX 加随后的 G-CSF 动员的 PBMCs（POI+PBMCs）和 CTX 加随后的 G-CSF 动员的 PBMCs 与 PRP（POI+PBMCs+PRP）。CTX 暴露诱导了典型的 POI 表型，表现为动情周期延长；动情期缩短；所有阶段的卵泡停滞；血清雌二醇-17β（E2）和抗苗勒管激素（AMH）水平降低；卵泡刺激素（FSH）水平升高。与 POI 大鼠相比，PRP 与动员的 PBMCs 一起移植导致动情周期、性激素水平和窦前卵泡生长更早恢复。在移植后 5、10 和 20 天，POI+PBMCs+PRP 雌性受体大鼠的卵巢组织中出现了雄性特异性 Sry 基因的表达，并伴随着血管生成标志物 CD34+和 VEGF 以及卵泡发生标志物 AMH 和 FSHR 的表达显著增加。此外，PBMCs 与 PRP 联合使用通过下调 BAX 和上调 BCL-2 减轻了颗粒细胞凋亡。这些结果表明，G-CSF 动员的 PBMCs 与 PRP 联合使用通过增加卵巢新生血管化、减少颗粒细胞凋亡和促进卵泡发生来加速 POI 大鼠卵巢功能的恢复。
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