Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism
Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism
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人羊膜间充质干细胞（hAD-MSC）移植至少部分通过旁分泌机制改善卵巢早衰（POI）大鼠的卵巢功能
DOI：
10.1186/s13287-019-1136-x
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发表时间：
2019-01-25
影响因子：
7.5
通讯作者：
Xiong, Zhengai
中科院分区：
文献类型：
--
作者：
Ling, Li;Feng, Xiushan;Xiong, Zhengai
背景：化疗可诱发卵巢早衰（POI），降低年轻女性患者的生育能力。目前，POI没有有效的治疗方法。人羊膜间充质干细胞（hAD-MSCs）可能是一种有前景的再生医学种子细胞。本研究探讨hAD-MSC移植对大鼠化疗诱导POI的影响及其机制。方法采用环磷酰胺腹腔注射法建立方案治疗型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组。用PKH26标记hAD-MSCs，并注入POI大鼠尾静脉。为了研究其潜在的机制，我们用免疫荧光染色分析了移植的hAD-MSCs在POI卵巢中的分化。采用ELISA法分析hAD-MSCs分泌的生长因子在hAD-MSCs条件培养基（hAD-MSCs - cm）中的体外表达。将60只雌性SD大鼠分为对照组、POI组和hAD-MSC-CM组，POI大鼠双侧卵巢注射hAD-MSC-CM。移植或注射hAD-MSC- cm后，检测血清性激素水平、发情周期、卵巢病理变化、卵泡计数、颗粒细胞（GC）凋亡及卵巢Bcl-2、Bax、VEGF表达。结果spkh26标记的hAD-MSCs在移植后主要归巢于卵巢。hAD-MSC移植可减轻POI大鼠卵巢损伤，改善卵巢功能。移植的hAD-MSCs仅位于卵巢间质，而非卵泡内，且不表达卵母细胞和GCs的典型标志物ZP3和FSHR。hAD-MSCs分泌FGF2、IGF-1、HGF和VEGF，在hAD-MSCs - cm中检测到这些生长因子。注射hAD-MSC-CM改善POI大鼠卵巢局部微环境，导致卵巢细胞Bax表达降低，Bcl-2和内源性VEGF表达升高，从而抑制化疗诱导的GC凋亡，促进血管生成，调节卵泡发育，从而部分减轻POI大鼠卵巢损伤，改善卵巢功能。结论shad - msc移植可部分通过旁分泌机制改善化疗诱导POI大鼠卵巢功能。hAD-MSCs介导的卵巢功能恢复存在旁分泌机制，可能归因于hAD-MSCs分泌的生长因子。
BackgroundChemotherapy 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 promisingseed cellfor regenerative medicine. This study investigated the effects and mechanisms of hAD-MSC transplantation on chemotherapy-induced POI in rats.MethodsChemotherapy-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.ResultsPKH26-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.ConclusionshAD-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.