hPMSC 移植通过 PI3K/Akt 信号通路恢复卵巢早衰小鼠卵巢功能与 Th17/Tc17 和 Th17/Treg 细胞比例的变化有关。
hPMSC transplantation restoring ovarian function in premature ovarian failure mice is associated with change of Th17/Tc17 and Th17/Treg cell ratios through the PI3K/Akt signal pathway.
机构信息
Department of Histology and Embryology, Binzhou Medical University, 346 Guanhai Rd, Yantai, Shandong, China.
Reproductive Medicine Center of the Affiliated Hospital of Binzhou Medical College, Binzhou, Shandong, China.
出版信息
Stem Cell Res Ther. 2018 Feb 14;9(1):37. doi: 10.1186/s13287-018-0772-x.
BACKGROUND
Human placenta-derived mesenchymal stem cell (hPMSC) transplantation has been demonstrated to be an effective way of recovering ovarian function in mice with autoimmune induced premature ovarian failure (POF). But the exact mechanism remains unclear. The goal of the present study is to investigate the role of immune factors (T-helper 17 (Th17), cytotoxic T (Tc17) and regulatory T (Treg) cells) in the recovery of ovarian function and whether the phosphatidylinositol 3-kinase (PI3K)/Akt signal pathway is involved in the regulation.
METHODS
The inhibitor of PI3K/Akt was administered to observe its effect on ovarian function recovery and immune regulation. Serum levels of estradiol (E), follicle stimulation hormone (FSH), luteinizing hormone (LH) and anti-Müllerian hormone (AMH)) and anti-Zona pellucida antibody (AZPAb) were measured by ELISA to evaluate ovarian function. The morphological changes of ovaries were observed by HE staining. Apoptosis of granular cells (GCs) was determined by detecting the expression of capase-3. Expression of p-Akt protein was detected by immunohistochemistry and western blot assay in ovarian tissues. The MTT assay was performed to assess GC proliferation. GC apoptosis was performed using flow cytometry analysis. Percentages of Th17, Tc17 and Treg cells were detected by flow cytometry. Expression of interleukin (IL)-17 in serum was measured by ELISA.
RESULTS
LY294002 administration decreased serum levels of E and AMH, while the levels of FSH, LH and AZPAb in serum were increased compared with mice in the hPMSC transplantation group. The ovarian morphology presented as atrophy and fibrosis, with functional follicles exhausted. The expression of p-Akt in ovarian tissue was significantly decreased. Also, LY294002 administration significantly decreased proliferation and increased cell apoptosis in GCs, and for immune factors the ratios of Th17/Tc17 and Th17/Treg cells were significantly increased, as well as the serum levels of IL-17.
CONCLUSIONS
Our data suggest that the PI3K/Akt signal pathway is involved in the recovery of ovarian function by changing the ratios of Th17/ Tc17 and Th17/Treg cells in POF mice following hPMSC transplantation.
背景
人胎盘间充质干细胞（hPMSC）移植已被证明是一种恢复自身免疫性诱导的卵巢早衰（POF）小鼠卵巢功能的有效方法。但确切的机制尚不清楚。本研究的目的是探讨免疫因子（辅助性 T 细胞 17（Th17）、细胞毒性 T（Tc17）和调节性 T（Treg）细胞）在卵巢功能恢复中的作用，以及是否涉及磷脂酰肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）信号通路的调节。
方法
给予 PI3K/Akt 抑制剂观察其对卵巢功能恢复和免疫调节的影响。采用酶联免疫吸附试验（ELISA）检测血清雌二醇（E）、卵泡刺激素（FSH）、黄体生成素（LH）和抗苗勒管激素（AMH）水平及抗透明带抗体（AZPAb）水平，评估卵巢功能。HE 染色观察卵巢形态学变化。采用 caspase-3 检测法检测颗粒细胞（GC）凋亡。采用免疫组织化学和蛋白质印迹法检测卵巢组织中 p-Akt 蛋白的表达。MTT 法检测 GC 增殖，流式细胞术分析 GC 凋亡，流式细胞术检测 Th17、Tc17 和 Treg 细胞的百分比。采用 ELISA 法检测血清白细胞介素（IL）-17 表达。
结果
与 hPMSC 移植组小鼠相比，LY294002 给药后血清 E 和 AMH 水平降低，而 FSH、LH 和 AZPAb 水平升高。卵巢形态表现为萎缩和纤维化，功能卵泡耗竭。卵巢组织中 p-Akt 的表达明显降低。此外，LY294002 给药后 GC 增殖明显减少，细胞凋亡增加，Th17/Tc17 和 Th17/Treg 细胞的比例以及血清 IL-17 水平也明显升高。
结论
我们的数据表明，PI3K/Akt 信号通路通过改变 hPMSC 移植后 POF 小鼠 Th17/Tc17 和 Th17/Treg 细胞的比例，参与卵巢功能的恢复。
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