Yousef Mohamed Samy, Akthar Ihshan, Ma Dongxue, Haneda Shingo, Kusama Kazuya, Shimada Masayuki, Imakawa Kazuhiko, Miyamoto Akio
Global Agromedicine Research Center (GAMRC), Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan.
Department of Theriogenology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt.
Front Immunol. 2025 Mar 31;16:1560996. doi: 10.3389/fimmu.2025.1560996. eCollection 2025.
Platelet derivatives improve the uterine immune environment and increase pregnancy success in humans and animals. Platelet-conditioned media (PCM) contain all molecules derived from platelets (platelet secretions). The present study aimed to investigate the immunological impacts of platelet secretions on polymorphonuclear neutrophils (PMNs) and bovine endometrial epithelial cells (BEECs), .
Platelets (10×10 platelets/mL) from Holstein dairy cows were incubated for 0.5 h or lysed to obtain the PCM and platelet lysate (Lysate), respectively. PMNs were stimulated with PCM for 3h. While BEECs were exposed to PCM or Lysate for 24 h. Real-time PCR was performed to detect the expression of targeted genes (cytokines), including , and . Lipoxin A4 (LXA4; anti-inflammatory mediator) and PGE2 concentrations in the supernatants of PMNs cultured with PCM were measured via ELISA. Cell proliferation in BEECs was assessed using the Cell Counting Kit-8 (CCK-8). Additionally, uterine explants were prepared and processed for immunofluorescence to determine the expression of the LXA4 receptor.
In PMNs, platelet secretions downregulated the mRNA expression of pro-inflammatory cytokines ( and ) and increased LXA4 production. In both PMNs and BEECs, platelet secretions upregulated expression and PGE2 production. In BEECs, platelet secretions and Lysate upregulated While Lysate suppressed mRNA expression. Further, platelet secretions showed an anti-proliferative effect in BEECs and increased the LXA4 receptor protein expression in the endometrial epithelia.
Our findings reveal for the first time that platelet secretions directly act on PMNs and BEECs , thereby assisting the uterine immune network to shift anti-inflammatory environment toward pregnancy. The present study can explain, in part, the successful applications of platelet derivatives in reproductive medicine.
血小板衍生物可改善子宫免疫环境，并提高人类和动物的妊娠成功率。血小板条件培养基（PCM）包含源自血小板的所有分子（血小板分泌物）。本研究旨在探讨血小板分泌物对多形核中性粒细胞（PMN）和牛子宫内膜上皮细胞（BEEC）的免疫影响。
将来自荷斯坦奶牛的血小板（10×10个血小板/毫升）分别孵育0.5小时或裂解以获得PCM和血小板裂解物（裂解物）。用PCM刺激PMN 3小时。而BEEC暴露于PCM或裂解物24小时。进行实时PCR以检测包括、和在内的靶向基因（细胞因子）的表达。通过ELISA测量用PCM培养的PMN上清液中脂氧素A4（LXA4；抗炎介质）和PGE2的浓度。使用细胞计数试剂盒-8（CCK-8）评估BEEC中的细胞增殖。此外，制备子宫外植体并进行免疫荧光处理以确定LXA4受体的表达。
在PMN中，血小板分泌物下调促炎细胞因子（和）的mRNA表达并增加LXA4的产生。在PMN和BEEC中，血小板分泌物均上调表达和PGE2的产生。在BEEC中，血小板分泌物和裂解物上调，而裂解物抑制mRNA表达。此外，血小板分泌物在BEEC中显示出抗增殖作用，并增加子宫内膜上皮中LXA4受体蛋白的表达。
我们的研究结果首次揭示血小板分泌物直接作用于PMN和BEEC，从而协助子宫免疫网络将抗炎环境转变为有利于妊娠的环境。本研究可以部分解释血小板衍生物在生殖医学中的成功应用。
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