Xu Rongxiang Regenerative Medicine Research Center, Binzhou Medical University, Yantai, 264003, Shandong, China.
College of Basic Medicine, Binzhou Medical University, Yantai, 264003, Shandong, China.
Stem Cell Rev Rep. 2023 Jul;19(5):1449-1465. doi: 10.1007/s12015-022-10469-y. Epub 2022 Oct 29.
The mechanism of human Umbilical Cord Mesenchymal Stem Cells (hUMSCs) transplantation to improve ovarian function in the rats with Premature Ovarian Insufficiency (POI) is still unclear. The aim of this study is to investigate the signal axis mechanism that is involved in the ovarian function recovery of POI rats following hUMSCs transplantation.
The rat model with POI was established by intraperitoneal injection of cisplatin. The hUMSCs were transplanted by caudal vein injection into POI rats. Hematoxylin-eosin (H&E) staining was performed to examine the morphology of rat ovarian tissue. Masson staining, Sirus red staining and immunofluorescence were used to observe the fibrosis extent of ovarian tissue. The levels of serum sex hormones and the expression of fibrosis related markers in ovarian tissues were measured by enzyme-linked immunosorbent assay (ELISA). The expression of NR4A1, Phospho-NR4A1 and AMP-activated protein kinase (AMPK) signaling in rat ovarian tissues was measured by immunohistochemistry and immunofluorescence. The role of AMPK/NR4A1 signaling axis in the regulation of ovarian function recovery in POI rats following hUMSCs transplantation was further investigated by adenovirus and siRNA intervention in isolated stromal cells.
The results showed that the hUMSCs transplantation significantly inhibited ovarian tissue fibrosis and restored the ovarian function in POI rats. The level of NR4A1 and AMPK expression in ovarian tissue of POI rats after hUMSCs transplantation was significantly increased compared with the control group. In the cultured ovarian stromal cells, the similar results were obtained on the expression of NR4A1 and its regulation on fibrosis related molecular markers in Cisplatin (CDDP) damaged stromal cells following hUMSCs supernatant treatment. Both hUMSCs supernatant treatment and the addition of AMPK inhibitors increased NR4A1 expression in stromal cells. And after NR4A1 molecular intervention, fibrosis-related indicators in stromal cells changed. The data suggests that the AMPK/NR4A1 signaling axis is involved in the ovarian function changes in POI rats following hUMSCs transplantation.
The data from this study indicate that the inhibition of tissue fibrosis and recovery of ovarian function is regulated by AMPK/NR4A1 signaling axis in POI rats following hUMSCs transplantation.
人脐带间充质干细胞（hUMSC）移植改善卵巢早衰（POI）大鼠卵巢功能的机制尚不清楚。本研究旨在探讨 hUMSC 移植后改善 POI 大鼠卵巢功能恢复涉及的信号轴机制。
采用顺铂腹腔注射建立 POI 大鼠模型，尾静脉注射 hUMSC 移植。苏木精-伊红（H&E）染色观察大鼠卵巢组织形态。Masson 染色、天狼猩红染色和免疫荧光观察卵巢组织纤维化程度。酶联免疫吸附试验（ELISA）测定血清性激素水平及卵巢组织纤维化相关标志物表达。免疫组化和免疫荧光法测定大鼠卵巢组织中 NR4A1、磷酸化 NR4A1 和 AMP 激活蛋白激酶（AMPK）信号的表达。腺病毒和 siRNA 干预分离的基质细胞进一步研究 AMPK/NR4A1 信号轴在 hUMSC 移植后调节 POI 大鼠卵巢功能恢复中的作用。
hUMSC 移植显著抑制卵巢组织纤维化，恢复 POI 大鼠卵巢功能。与对照组相比，hUMSC 移植后 POI 大鼠卵巢组织中 NR4A1 和 AMPK 表达水平明显升高。在培养的卵巢基质细胞中，hUMSC 上清液处理 Cisplatin（CDDP）损伤的基质细胞后，NR4A1 的表达及其对纤维化相关分子标志物的调节作用也得到了类似的结果。hUMSC 上清液处理和添加 AMPK 抑制剂均增加了基质细胞中 NR4A1 的表达。NR4A1 分子干预后，基质细胞中纤维化相关指标发生变化。数据表明，AMPK/NR4A1 信号轴参与 hUMSC 移植后 POI 大鼠卵巢功能变化。
本研究数据表明，AMPK/NR4A1 信号轴调节 hUMSC 移植后 POI 大鼠组织纤维化抑制和卵巢功能恢复。
