Human amniotic mesenchymal stem cells improve ovarian function in natural aging through secreting hepatocyte growth factor and epidermal growth factor.
Human amniotic mesenchymal stem cells improve ovarian function in natural aging through secreting hepatocyte growth factor and epidermal growth factor.
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DOI：
10.1186/s13287-018-0781-9
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发表时间：
2018-03-09
影响因子：
7.5
通讯作者：
Huang B
中科院分区：
文献类型：
--
作者：
Ding C;Zou Q;Wang F;Wu H;Chen R;Lv J;Ling M;Sun J;Wang W;Li H;Huang B
虽然许多报道表明，各种干细胞在卵巢早衰中具有恢复功能的能力，但很少有研究关注干细胞治疗卵巢自然衰老(NOA)。我们设计了这项实验研究来研究人羊膜间充质干细胞(HAMSCs)是否保留了恢复卵巢功能的能力，以及hAMSCs在这一过程中是如何工作的。建立NOA小鼠模型，正常饲养12~14个月，3~5月龄雌性幼鼠为正常对照组。苏木精-伊红染色可进行卵泡计数并显示卵巢组织结构。采用酶联免疫吸附试验检测血清性激素雌二醇(E_2)、抗苗勒氏激素(AMH)和卵泡刺激素(FSH)水平。采用流式细胞术(FACS)检测人卵巢颗粒细胞(HGC)(Ki67、AMH、FSH受体、FOXL2和CYP19A1)的增殖率和标志物表达水平。用蛋白抗体阵列方法检测细胞因子(生长因子)。肝细胞生长因子(HGF)和表皮生长因子(EGF)与hGCs共培养后，用流式细胞仪检测细胞增殖(Ki67)和凋亡(Annexin V)水平。将HGF和EGF注入自然衰老小鼠的卵巢，检测卵泡总数和激素水平。HAMSCs移植到NOA小鼠模型后，通过分四个阶段改善卵泡数，对小鼠卵巢功能起到治疗作用。此外，我们的结果显示hAMSCs显著促进NOA患者卵巢颗粒细胞的增殖率和标志物的表达水平。同时，我们发现hAMSCs分泌的EGF和HGF水平高于其他生长因子。联合应用生长因子(HGF和EGF)可显著提高NOA模型小鼠卵巢细胞的增殖率和抑制凋亡率，卵巢内注射HGF和EGF后卵泡总数和激素水平均恢复到正常水平。这些发现为hAMSC在抵抗NOA中发挥不可或缺的作用这一概念提供了洞察力。此外，我们目前的研究表明，来源于hAMSCs的生长因子组合在抑制卵巢衰老方面发挥着核心作用。因此，我们认为hAMSCs通过分泌HGF和EGF来改善自然衰老条件下的卵巢功能。本文的在线版本(10.1186/s13287-0180781-9)包含补充材料，可供授权用户使用。
Although many reports show that various kinds of stem cells have the ability to recover function in premature ovarian aging, few studies have looked at stem cell treatment of natural ovarian aging (NOA). We designed this experimental study to investigate whether human amniotic mesenchymal stem cells (hAMSCs) retain the ability to restore ovarian function, and how hAMSCs work in this process. To build the NOA mouse model, the mice were fed for 12–14 months normally with young fertile female mice as the normal control group (3–5 months old). Hematoxylin and eosin staining permitted follicle counting and showed the ovarian tissue structure. An enzyme-linked immunosorbent assay was used to detect the serum levels of the sex hormones estradiol (E2), anti-mullerian hormone (AMH), and follicle-stimulating hormone (FSH). The proliferation rate and marker expression level of human ovarian granule cells (hGCs) (ki67, AMH, FSH receptor, FOXL2, and CYP19A1) were measured by flow cytometry (FACS). Cytokines (growth factors) were measured by a protein antibody array methodology. After hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were co-cultured with hGCs, proliferation (ki67) and apoptosis (Annexin V) levels were analyzed by FACS. After HGF and EGF were injected into the ovaries of natural aging mice, the total follicle numbers and hormone levels were tested. After the hAMSCs were transplanted into the NOA mouse model, the hAMSCs exerted a therapeutic activity on mouse ovarian function by improving the follicle numbers over four stages. In addition, our results showed that hAMSCs significantly promoted the proliferation rate and marker expression level of ovarian granular cells that were from NOA patients. Meanwhile, we found that the secretion level of EGF and HGF from hAMSCs was higher than other growth factors. A growth factor combination (HGF with EGF) improved the proliferation rate and inhibited the apoptosis rate more powerfully after a co-culture with hGCs, and total follicle numbers and hormone levels were elevated to a normal level after the growth factor combination was injected into the ovaries of the NOA mouse model. These findings provide insight into the notion that hAMSCs play an integral role in resistance to NOA. Furthermore, our present study demonstrates that a growth factor combination derived from hAMSCs plays a central role in inhibiting ovarian aging. Therefore, we suggest that hAMSCs improve ovarian function in natural aging by secreting HGF and EGF. The online version of this article (10.1186/s13287-018-0781-9) contains supplementary material, which is available to authorized users.
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影响因子：
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作者：
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通讯作者：
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影响因子：
6.1
作者：
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通讯作者：
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