Cheng Keyan, Cui Jialei, Zhou Wenli, Liu Huiqiang, Yang Taotao, Wang Yonghong
Department of Gynecology and Obstetrics, The Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
Matern Fetal Med. 2022 Sep 27;4(4):238-244. doi: 10.1097/FM9.0000000000000169. eCollection 2022 Oct.
The objective of this study was to investigate the expression levels of microRNA-141-5p(miRNA-141-5p), MAPK1 and neutrophil elastase in patients with and without preeclampsia (PE), and the relationship between miRNA-141-5p and MAPK1 with respect to the secretion of elastase by neutrophils in patients with PE.
Thirty patients with PE and 30 healthy pregnant (HP) women were recruited from The Second Hospital of Shanxi Medical University, Taiyuan, China, between February 2017 and July 2018. Neutrophils were isolated from 8 mL peripheral blood samples and cultured. We recorded neutrophil count and morphology during culture. Apoptosis was detected by flow cytometry in different groups at 0, 24, and 48 h. The expression levels of elastase were detected in neutrophils by enzyme-linked immunosorbent assay, whereas the expression levels of miRNA-141-5p in peripheral blood neutrophils were detected by real-time polymerase chain reaction. We used TargetScanHuman Release 7.2 to analyze the target genes of miRNA-141-5p. The expression of MAPK1 in peripheral blood neutrophils was detected by western blotting. Data were analyzed by SPSS version 21.0 software, and comparisons between groups were carried out with the Student test.
There was no significant difference between the PE and HP groups ( > 0.050) with regard to age or body mass index. The weight of newborns in the PE group (2846.00 ± 600.00 g) was significantly lower than that in the HP group (3055.00 ± 230.68 g). The number of neutrophilic granulocytes(NGs) in blood samples from the PE group was significantly higher than that in the HP group ( = 0.003). There was no significant difference between the groups with regard to morphology. Apoptosis in the PE group was delayed when compared with the HP group at different time points. The value of apoptosis in the PE and HP groups were respectively 0.790, < 0.001 and 0.030 at 0 h, 24 h and 48 h. The expression levels of miRNA-141-5p in the PE group were significantly lower than those in the HP group ( < 0.050). The expression levels of MAPK1 in neutrophils from the PE group were significantly higher than those in the HP group ( < 0.050) by western blot. The expression levels of elastase in neutrophils from the PE group were significantly higher than those in the HP group ( < 0.050). Furthermore, the number of NGs in peripheral blood from the PE group was higher than that of the HP group; however, the levels of apoptosis were lower. The expression levels of miRNA-141-5p in NGs decreased, the expression of MAPK1 increased, and the secretion of neutrophil elastase in the NG medium increased in the PE group than those in the HP group.
Collectively, our analysis suggested that miRNA-141-5p may be involved in the pathogenesis of PE by regulating the MAPK1 signaling pathway to activate neutrophils and increase the secretion of elastase.
本研究旨在调查子痫前期（PE）患者和未患子痫前期患者中微小RNA - 141 - 5p（miRNA - 141 - 5p）、丝裂原活化蛋白激酶1（MAPK1）和中性粒细胞弹性蛋白酶的表达水平，以及在子痫前期患者中miRNA - 141 - 5p和MAPK1与中性粒细胞弹性蛋白酶分泌之间的关系。
2017年2月至2018年7月期间，从中国山西省太原市山西医科大学第二医院招募了30例子痫前期患者和30例健康孕妇（HP）。从8 mL外周血样本中分离并培养中性粒细胞。我们记录了培养过程中的中性粒细胞计数和形态。在0、24和48小时通过流式细胞术检测不同组的细胞凋亡情况。通过酶联免疫吸附测定法检测中性粒细胞中弹性蛋白酶的表达水平，而通过实时聚合酶链反应检测外周血中性粒细胞中miRNA - 141 - 5p的表达水平。我们使用TargetScanHuman Release 7.2分析miRNA - 141 - 5p的靶基因。通过蛋白质印迹法检测外周血中性粒细胞中MAPK1的表达。数据采用SPSS 21.0软件进行分析，组间比较采用Student检验。
子痫前期组和健康孕妇组在年龄或体重指数方面无显著差异（P>0.050）。子痫前期组新生儿体重（2846.00±600.00 g）显著低于健康孕妇组（3055.00±230.68 g）。子痫前期组血样中的中性粒细胞（NGs）数量显著高于健康孕妇组（P = 0.003）。两组在形态方面无显著差异。在不同时间点，子痫前期组的细胞凋亡与健康孕妇组相比延迟。子痫前期组和健康孕妇组在0小时、24小时和48小时的细胞凋亡P值分别为0.790、<0.001和0.030。子痫前期组中miRNA - 141 - 5p的表达水平显著低于健康孕妇组（P<0.050）。通过蛋白质印迹法检测，子痫前期组中性粒细胞中MAPK1的表达水平显著高于健康孕妇组（P<0.050）。子痫前期组中性粒细胞中弹性蛋白酶的表达水平显著高于健康孕妇组（P<0.050）。此外，子痫前期组外周血中的中性粒细胞数量高于健康孕妇组；然而，细胞凋亡水平较低。与健康孕妇组相比，子痫前期组中性粒细胞中miRNA - 141 - 5p的表达水平降低，MAPK1的表达增加，中性粒细胞培养基中中性粒细胞弹性蛋白酶的分泌增加。
总体而言，我们的分析表明，miRNA - 141 - 5p可能通过调节MAPK1信号通路激活中性粒细胞并增加弹性蛋白酶的分泌，参与子痫前期的发病机制。
