富血小板血浆促进人子宫内膜间充质干细胞（EnMSCs）增殖的机制

王晓寒; 牟善茂; 郝翠芳; 任琳琳; 王敏; 赵彤

doi:10.12259/j.issn.2095-610X.S20220126

富血小板血浆促进人子宫内膜间充质干细胞（EnMSCs）增殖的机制

doi: 10.12259/j.issn.2095-610X.S20220126

王晓寒¹,
牟善茂²,
郝翠芳^{3, 4},
任琳琳¹,
王敏¹,
赵彤^1, ,

1.
日照市中医医院产科
2.
脑病科，山东日照　276800
3.
青岛大学附属青岛妇女儿童医院生殖中心，山东青岛　266011
4.
青岛大学附属烟台毓璜顶医院生殖中心，山东烟台　264000

基金项目: 山东省医药卫生科技发展计划基金资助项目（202005031192）

详细信息

作者简介:
王晓寒（1983～），女，山东泰安人，医学博士，主治医师，主要从事子宫内膜容受性及女性生殖医学研究工作

通讯作者:
赵彤，E-mail： zhaotong667@163.com

中图分类号: R714
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出版历程
- 收稿日期: 2021-12-18
- 刊出日期: 2022-01-25

Platelet-rich Plasma Promotes the Proliferation of Human Endometrial Mesenchymal Stem Cells (EnMSCs) through the PI3K/AKT/mTOR Signaling Pathway

1.
Dept. of Obstetrics
2.
Dept. of Encephalopathy，Rizhao Hospital of Traditional Chinese Medicine，Rizhao Shandong 276800
3.
Center for Reproductive Medicine，Qingdao Women and Children’s Hospital，Qingdao University，Qingdao Shandong 266011
4.
Center for Reproductive Medicine，Yantai Yuhuangding Hospital Affiliated to Qingdao University， Yantai Shandong 264000，China

摘要

摘要: 目的探讨PRP促进EnMSCs增殖的机制，为EnMSCs的扩增及临床应用提供理论基础。方法将EnMSCs随机分为对照组、2%PRP组、2%PRP + LY294002组，CCK-8法检测细胞增殖情况；流式细胞仪监测细胞周期及Western Blot和ELISA检测细胞中p-PI3K、AKT、p-AKT、mTOR及p-mTOR的蛋白表达。结果（1） CCK-8结果显示：与对照组相比较，2%PRP组EnMSCs的增殖水平显著较高(P < 0.05)；与2%PRP组相比，2%PRP+LY294002组EnMSCs的增殖水平显著较低（ P < 0.05）；（2）流式细胞仪分析细胞周期结果显示：2%PRP组G2/M期细胞比例显著高于对照组和2%PRP+ LY294002组，G0/G1细胞比例明显低于对照组和2%PRP+ LY294002组，差异均有统计学意义（ P < 0.05）；（3） 2%PRP组EnMSCs中p-PI3K、AKT、p-AKT、mTOR、p-mTOR的蛋白表达明显高于对照组及2%PRP+LY294002组，差异均有统计学意义（ P < 0.05）。结论 EnMSCs的增殖由PI3K/AKT/ mTOR信号通路调控，PRP通过促进AKT、mTOR的蛋白表达及其磷酸化以激活该通路，促进EnMSCs由G1期向G2期转变，从而促进EnMSCs的增殖。
- 富血小板血浆 /
- 子宫内膜间充质干细胞 /
- PI3K/AKT/mTOR /
- 细胞增殖 /
- 细胞周期
Abstract: Objective To explore the mechanism of PRP promoting EnMSCs proliferation, and to provide a theoretical basis for the expansion and clinical application of EnMSCs. Methods EnMSCs were randomly divided into the control group, 2%PRP group and 2%PRP + LY294002 group. Cell proliferation was detected by CCK-8 method. Cell cycle was monitored by flow cytometry and protein expressions of p-PI3K, AKT, p-AKT, mTOR and p-mTOR were detected by Western Blot. Results （1） CCK-8 results: Compared with the control group, the proliferation level of EnMSCs in 2%PRP group was significantly higher （P < 0.05）. Compared with the 2%PRP group, the proliferation level of EnMSCs in the 2%PRP+LY294002 group was significantly lower （ P < 0.05）. （2） The results of cell cycle analysis by flow cytometry showed that the proportion of cells in G2/M phase in the 2% PRP group was significantly higher than that in the control group and 2% PRP+LY294002 group, and the proportion of cells in G0/G1 phase was significantly lower than that in the control group and 2% PRP+ LY294002 group. All of these had statistically significant differences （ P < 0.05）. （3） The protein expressions of p-PI3K, AKT, p-AKT, mTOR and p-mTOR in EnMSCs in the 2%PRP group were significantly higher than those in the control group and the 2%PRP+LY294002 group, with statistical significance （ P < 0.05）. Conclusion The proliferation of EnMSCs is regulated by the PI3K/Akt/mTOR signaling pathway. PRP activates this pathway by promoting the protein expression and phosphorylation of AKT and mTOR, thereby promoting the proliferation of EnMSCs.
- Platelet-rich plasma /
- Endometrial mesenchymal stem cells /
- PI3K/AKT/mTOR /
- Cell proliferation /
- Cell cycle.

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图 1 各组48h EnMSCs生长情况（×200）

A：对照组；B：2%PRP组；C：2%PRP + LY294002组

Figure 1. The 48 h EnMSCs proliferation phenomena for each group （×200）

下载: 全尺寸图片幻灯片

图 2 3组CCK-8增殖结果比较

与对照组比较，^**P < 0.01。

Figure 2. The CCK-8 proliferation results comparison between the three groups

下载: 全尺寸图片幻灯片

图 3 各组流式细胞周期及细胞周期比例图

A：各组流式细胞周期图；B：细胞周期比例图。

Figure 3. The flow cell cycle and cell cycle proportion gram in three groups

下载: 全尺寸图片幻灯片

图 4 3组p-PI3K、AKT、p-AKT、mTOR、p-mTOR蛋白表达水平比较

A：Western blot结果；B：ELISA结果。

Figure 4. The p-PI3K, AKT, p-AKT, mTOR, and p-mTOR proteins demonstration levels comparison between the three groups

下载: 全尺寸图片幻灯片

表 1 3组EnMSCs细胞周期比例（%）比较（n = 3）

Table 1. The EnMSCs cell cycle proportion comparison in the three groups （n = 3）

组别	G1/G0（%）	S（%）	G2/M（%）
对照组	70.04 ± 2.67	25.49 ± 2.68	4.47 ± 1.04
2%PRP组	54.63 ± 4.23^**▲	30.66 ± 2.85^*▲	14.71 ± 2.72^**▲
2%PRP+ LY294002组	72.16 ± 4.26	17.45 ± 3.49^**	10.39 ± 2.84^**
与对照组比较，^P < 0.05, ^*P < 0.01；与2%PRP+ LY294002组比较， ^▲P < 0.01。

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