Role of B7H4 and Fas in regulation of trophoblast cells and development of pre-eclampsia: a cross-sectional study

This study aimed to investigate the role of B7H4 and Fas in pre-eclampsia (PE) occurrence and development and reveal its potential mechanisms. Thirty healthy individuals and 60 patients with PE were enrolled in the study. In addition, the clinical characteristics of all participants were collected, including age, gestational weeks at delivery, gestational time, number of births, systolic blood pressure, diastolic blood pressure and foetal weight. The chi-square test was used to evaluate differences in clinical characteristics between the high- and low-expression groups. The expression levels of B7H4 and Fas were analysed using western blotting and real-time quantitative polymerase chain reaction (RT-qPCR). The upstream regulators of B7H4 in trophoblasts were predicted and estimated using a luciferase reporter assay. The proliferation and motility of trophoblasts were assessed using CCK8 and transwell assays, respectively. B7H4 and Fas were upregulated in PE (<i>p</i> &lt; 0.05) and showed diagnostic potential with insufficient sensitivity and specificity [B7H4: area under curve (AUC) = 0.790, sensitivity = 65%, specificity = 83.33%; Fas: AUC = 0.717, sensitivity = 68.34%, specificity = 73.33%]. B7H4 and Fas were closely associated with increased blood pressure in patients with PE (<i>p</i> &lt; 0.05), and the combination of B7H4 and Fas increased the diagnostic efficacy (AUC = 0.864), sensitivity (73.33%) and specificity (86.67%). In trophoblast cells, miR-4319 negatively regulated B7H4 and Fas expression as well as cell proliferation, migration and invasion (<i>p</i> &lt; 0.05). Overexpression of B7H4 alleviated the inhibitory effects of miR-4319, which were reversed by Fas knockdown (<i>p</i> &lt; 0.05). miR-4319 mediates the progression of trophoblast progression by negatively regulating the expression of B7H4 and Fas. Therefore, B7H4 and Fas may serve as potential biomarkers for the prediction and treatment of PE. This study aimed to evaluate the significance of B7H4 and Fas in the occurrence and development of pre-eclampsia (PE) and to reveal its potential mechanism. We found that the upregulated B7H4 and Fas served as diagnostic biomarkers for PE, and their combination showed higher diagnostic efficacy, sensitivity and specificity. miR-4319 negatively regulates B7H4 and Fas in trophoblasts and therefore mediates cell proliferation, migration and invasion.

本研究旨在探讨B7H4与Fas在子痫前期（pre-eclampsia, PE）发生发展中的作用，并揭示其潜在分子机制。本研究共招募30名健康个体与60名PE患者作为研究对象，收集所有受试者的临床特征数据，包括年龄、分娩孕周、妊娠时长、分娩次数、收缩压、舒张压及胎儿体重。采用卡方检验（chi-square test）分析高低表达组间临床特征的差异。通过蛋白质印迹法（Western Blot）与实时定量聚合酶链反应（real-time quantitative polymerase chain reaction, RT-qPCR）检测B7H4与Fas的表达水平。利用荧光素酶报告基因实验（luciferase reporter assay）预测并验证滋养层细胞（trophoblasts）中B7H4的上游调控因子。分别采用CCK8实验与Transwell实验评估滋养层细胞的增殖能力与迁移侵袭能力。
B7H4与Fas在PE患者体内呈高表达（*P* < 0.05），且二者均具备一定诊断价值，但敏感性与特异性均存在不足[B7H4：曲线下面积（area under curve, AUC）=0.790，敏感性=65%，特异性=83.33%；Fas：AUC=0.717，敏感性=68.34%，特异性=73.33%]。B7H4与Fas的表达水平与PE患者的血压升高显著相关（*P* < 0.05），二者联合检测可提升诊断效能（AUC=0.864，敏感性73.33%，特异性86.67%）。
在滋养层细胞中，miR-4319（microRNA-4319）可负向调控B7H4与Fas的表达，同时抑制细胞增殖、迁移与侵袭能力（*P* < 0.05）。过表达B7H4可缓解miR-4319介导的抑制效应，而该效应可通过敲低Fas得以逆转（*P* < 0.05）。miR-4319通过负向调控B7H4与Fas的表达，介导滋养层细胞的增殖、迁移与侵袭过程。因此，B7H4与Fas或可作为PE预测与治疗的潜在生物标志物。
本研究旨在评估B7H4与Fas在子痫前期（PE）发生发展中的意义，并揭示其潜在分子机制。本研究发现，高表达的B7H4与Fas可作为PE的诊断生物标志物，二者联合检测可获得更高的诊断效能、敏感性与特异性。miR-4319可在滋养层细胞中负向调控B7H4与Fas的表达，进而介导细胞的增殖、迁移与侵袭过程。