Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Chongqing Medical University, No. 76, Linjiang Road, Chongqing, 400010, China.
State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and the Ministry of Science and Technology, Chongqing Key Laboratory of Biomedical Engineering, College of Biomedical Engineering, Chongqing Medical University, Chongqing, 400010, China.
Stem Cell Res Ther. 2019 Jan 25;10(1):46. doi: 10.1186/s13287-019-1136-x.
Chemotherapy can induce premature ovarian insufficiency (POI) and reduce fertility in young female patients. Currently, there is no effective therapy for POI. Human amnion-derived mesenchymal stem cells (hAD-MSCs) may be a promising seed cell for regenerative medicine. This study investigated the effects and mechanisms of hAD-MSC transplantation on chemotherapy-induced POI in rats.
Chemotherapy-induced POI rat models were established by intraperitoneal injection of cyclophosphamide. Seventy-two female SD rats were randomly divided into control, POI, and hAD-MSC-treated groups. hAD-MSCs were labeled with PKH26 and injected into the tail veins of POI rats. To examine the underlying mechanisms, the differentiation of transplanted hAD-MSCs in the POI ovaries was analyzed by immunofluorescent staining. The in vitro expression of growth factors secreted by hAD-MSCs in hAD-MSC-conditioned media (hAD-MSC-CM) was analyzed by ELISA. Sixty female SD rats were divided into control, POI, and hAD-MSC-CM-treated groups, and hAD-MSC-CM was injected into the bilateral ovaries of POI rats. After hAD-MSC transplantation or hAD-MSC-CM injection, serum sex hormone levels, estrous cycles, ovarian pathological changes, follicle counts, granulosa cell (GC) apoptosis, and Bcl-2, Bax, and VEGF expression in ovaries were examined.
PKH26-labeled hAD-MSCs mainly homed to ovaries after transplantation. hAD-MSC transplantation reduced ovarian injury and improved ovarian function in rats with POI. Transplanted hAD-MSCs were only located in the interstitium of ovaries, rather than in follicles, and did not express the typical markers of oocytes and GCs, which are ZP3 and FSHR, respectively. hAD-MSCs secreted FGF2, IGF-1, HGF, and VEGF, and those growth factors were detected in the hAD-MSC-CM. hAD-MSC-CM injection improved the local microenvironment of POI ovaries, leading to a decrease in Bax expression and an increase in Bcl-2 and endogenous VEGF expression in ovarian cells, which inhibited chemotherapy-induced GC apoptosis, promoted angiogenesis and regulated follicular development, thus partly reducing ovarian injury and improving ovarian function in rats with POI.
hAD-MSC transplantation can improve ovarian function in rats with chemotherapy-induced POI at least partly through a paracrine mechanism. The presence of a paracrine mechanism accounting for hAD-MSC-mediated recovery of ovarian function might be attributed to the growth factors secreted by hAD-MSCs.
化疗会导致年轻女性患者发生卵巢早衰（POI）并降低生育能力。目前，POI 尚无有效的治疗方法。人羊膜间充质干细胞（hAD-MSCs）可能是再生医学有前途的种子细胞。本研究探讨了 hAD-MSC 移植对化疗诱导的 POI 大鼠的影响及其机制。
通过腹腔注射环磷酰胺建立化疗诱导的 POI 大鼠模型。将 72 只雌性 SD 大鼠随机分为对照组、POI 组和 hAD-MSC 治疗组。用 PKH26 标记 hAD-MSCs，然后将其注入 POI 大鼠的尾静脉。为了研究潜在机制，通过免疫荧光染色分析移植到 POI 卵巢中的 hAD-MSCs 的分化情况。通过 ELISA 分析 hAD-MSC 条件培养基（hAD-MSC-CM）中 hAD-MSCs 分泌的生长因子的表达。将 60 只雌性 SD 大鼠分为对照组、POI 组和 hAD-MSC-CM 治疗组，然后将 hAD-MSC-CM 注入 POI 大鼠的双侧卵巢。在 hAD-MSC 移植或 hAD-MSC-CM 注射后，检测血清性激素水平、动情周期、卵巢病理变化、卵泡计数、颗粒细胞（GC）凋亡以及卵巢中 Bcl-2、Bax 和 VEGF 的表达。
PKH26 标记的 hAD-MSCs 移植后主要归巢到卵巢。hAD-MSC 移植减轻了 POI 大鼠的卵巢损伤并改善了其卵巢功能。移植的 hAD-MSCs 仅位于卵巢间质中，而不在卵泡中，并且不表达卵母细胞和 GC 的典型标志物，即 ZP3 和 FSHR。hAD-MSCs 分泌 FGF2、IGF-1、HGF 和 VEGF，这些生长因子在 hAD-MSC-CM 中被检测到。hAD-MSC-CM 注射改善了 POI 卵巢的局部微环境，导致卵巢细胞中 Bax 表达减少，Bcl-2 和内源性 VEGF 表达增加，从而抑制了化疗诱导的 GC 凋亡，促进了血管生成并调节了卵泡发育，因此部分减轻了 POI 大鼠的卵巢损伤并改善了其卵巢功能。
hAD-MSC 移植至少部分通过旁分泌机制改善了化疗诱导的 POI 大鼠的卵巢功能。hAD-MSC 介导的卵巢功能恢复的旁分泌机制可能归因于 hAD-MSCs 分泌的生长因子。
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