甲酰肽受体2通过p38 MAPK通路参与复发性流产的发生

doi:10.16098/j.issn.0529-1356.2022.06.013

摘要/Abstract

摘要：

目的探讨甲酰肽受体2(FPR2)在复发性流产患者（RSA）绒毛组织中的表达，及其对滋养细胞增殖、迁移和侵袭的影响及作用机制。明确FPR2对滋养细胞功能的影响，分析其在复发性流产中的作用，并对机制进行探讨。方法收集临床样本（正常30例，RSA30例），利用免疫组织化学染色、Real-time PCR和Western blotting等方法，分析FPR2在RSA患者和正常绒毛组织间的定位和表达差异；应用CRISPR/Cas-9技术，对人绒毛膜滋养细胞系HTR-8/Svneo的FPR2进行敲降并验证；采用CCK-8实验、划痕实验、Transwell实验检测FPR2敲降后滋养细胞增殖、迁移和侵袭能力改变；单独或联合使用p38 MAPK抑制剂SB203580后，采用免疫荧光染色和Western blotting分析FPR2敲降后磷酸化的p38 MAPK（p-p38 MAPK）及p38 MAPK的表达水平；采用CCK-8实验、划痕实验、Transwell实验检测滋养细胞增殖、迁移和侵袭能力改变。结果复发性流产患者绒毛组织FPR2表达增加；FPR2敲低显著提高了HTR-8/Svneo细胞增殖、迁移和侵袭能力。FPR2敲低显著上调p-p38 MAPK表达水平，而加入SB203580抑制p38 MAPK通路后，FPR2敲降对滋养细胞的迁移和侵袭能力增强作用被部分逆转。结论 FPR2在RSA 患者绒毛滋养细胞中高表达，并通过p38 MAPK信号通路抑制滋养细胞的迁移、侵袭，可能在复发性流产发生中发挥重要作用。

关键词: 甲酰肽受体2, 复发性流产, 滋养细胞, p38 MAPK信号通路, 免疫荧光, 免疫印迹法, 人

Abstract:

Objective To explore the express of formyl peptide receptor 2(FPR2) in villi of recurrent spontaneous abortion (RSA), the effect on proliferation, migration and invasion of trophoblast, and the mechanism to clarify the effect of FPR2 on trophoblast function and explore its role and mechanism in recurrent spontaneous abortion. Methods Clinical villus specimens of 30 normal and 30 RSA patients were collected. Immunohistochemical staining, Real-time PCR and Western blotting were used to detect the location and expression of FPR2 in villi of patients with RSA and normal pregnant women. CRISPR/Cas-9 technique was used to knock down FPR2 in HTR-8/SVneo cells, CCK-8 assay, wound healing and Transwell assays were used to determine the ability of cell viability, migration and invasion. Immunofluorescent staining and Western blotting were used to analyze the changes of phosphorylated p38 MAPK(p-p38 MAPK)/p38 MAPK protein expression after applying with p38 MAPK inhibitor SB203580 alone or in combination. Results The expression of FPR2 in villi of patients with RSA increased. FPR2 knock-down improved the biological functions of HTR-8/Svneo cells such as proliferation, migration and invasion significantly. The expression of p-p38 MAPK was up-regulated significantly by FPR2 knock-down, and the ability enhancement of migration and invasion of trophoblasts was reversed partially by SB203580 which inhibits p38 MAPK pathway. FPR2 knock-down caused the change of p38 MAPK signaling pathway related to proteins. Conclusion FPR2 is highly expressed in trophoblasts of RSA patients, and inhibits the migration and invasion of trophoblasts through p38 MAPK signaling pathway, which may play an important role in RSA.

Key words: Formyl peptide receptor 2, Recurrent spontaneous abortion, Trophoblast, p38 MAPK signaling pathway, Immunofluorescence, Western blotting, Human

中图分类号:

R714.21

李安娜房振亚周美娟李淑贤赵曼郭君君张美华. 甲酰肽受体2通过p38 MAPK通路参与复发性流产的发生[J]. 解剖学报, 2022, 53(6): 785-792.

LI An-na FANG Zhen-ya ZHOU Mei-juan LI Shu-xian ZHAO Man GUO Jun-jun ZHANG Mei-hua. Effect of formyl peptide receptor 2 in recurrent spontaneous abortion through p38 MAPK pathway[J]. Acta Anatomica Sinica, 2022, 53(6): 785-792.

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