Department of Obstetrics and Gynecology, Taizhou Central Hospital (Taizhou University Hospital), China.
Department of Obstetrics and Gynecology, Taizhou Central Hospital (Taizhou University Hospital), China.
Reprod Biol. 2022 Dec;22(4):100696. doi: 10.1016/j.repbio.2022.100696. Epub 2022 Oct 31.
Preeclampsia (PE) is a serious obstetric complication, in which trophoblast cell invasion and migration contribute to placental inflammation. In line with the discovery that mRNA prostaglandin endoperoxide synthase 2 (PTGS2) participates in the inflammatory responses in various disorders, our study aims to explore the role of PTGS2 in trophoblast invasion and further in inflammatory response in PE, ultimately providing new therapeutic targets. Bioinformatics analysis was exploited to examine PTGS2 expression in GSE40182 and find inflammatory response-relevant genes in downstream targets of PTGS2. HTR-8/SVneo cells were treated with lipopolysaccharide (LPS) and transfected with short hairpin RNA against PTGS2 (shPTGS2). Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and immunofluorence assays were performed to quantify the expressions of PTGS2 and involved genes (matrix metallopeptidase 2 (MMP-2), tissue inhibitors of metalloproteinase-2 (TIMP-2), p65, p-p65, IκB-α, p-IκB-α, PTGIS, CAV1, AGTR1). The migration and invasion of trophoblasts were detected through wound healing and Transwell assays. We screened out PTGS2 from GSE40182 dataset. LPS promoted cell migration and invasion, the expressions of PTGS2 and MMP-2, and reduced the expression of TIMP-2, while PTGS2 knockdown reversed all above effects of LPS. Activation of nuclear factor kappa-B (NF-κB) pathway was reinforced by LPS which also upregulated CAV1 and AGTR1 levels, and downregulated PTGIS level. Also, the effects of LPS were offset by PTGS2 knockdown. Altogether, PTGS2 silencing reverses the promoting effect of LPS on trophoblast invasion and inflammation in PE, making a breakthrough in the research regarding molecular mechanism of PE.
子痫前期(PE)是一种严重的产科并发症,其中滋养细胞浸润和迁移有助于胎盘炎症。鉴于信使 RNA 前列腺素内过氧化物合酶 2(PTGS2)参与各种疾病的炎症反应的发现,我们的研究旨在探讨 PTGS2 在滋养细胞浸润中的作用,并进一步探讨其在 PE 中的炎症反应中的作用,最终为治疗提供新的靶点。利用生物信息学分析,在 GSE40182 中检查 PTGS2 的表达,并在 PTGS2 的下游靶点中找到与炎症反应相关的基因。用脂多糖(LPS)处理 HTR-8/SVneo 细胞,并转染针对 PTGS2 的短发夹 RNA(shPTGS2)。通过定量实时聚合酶链反应(qRT-PCR)、Western blot 和免疫荧光测定来量化 PTGS2 和涉及的基因(基质金属蛋白酶 2(MMP-2)、金属蛋白酶组织抑制剂 2(TIMP-2)、p65、p-p65、IκB-α、p-IκB-α、PTGIS、CAV1、AGTR1)的表达。通过划痕愈合和 Transwell 测定检测滋养细胞的迁移和侵袭。我们从 GSE40182 数据集筛选出 PTGS2。LPS 促进细胞迁移和侵袭,PTGS2 和 MMP-2 的表达减少,TIMP-2 的表达减少,而 PTGS2 敲低逆转了 LPS 的所有上述作用。核因子 kappa-B(NF-κB)通路的激活被 LPS 增强,同时也上调了 CAV1 和 AGTR1 的水平,并下调了 PTGIS 的水平。此外,LPS 的作用被 PTGS2 敲低抵消。总之,PTGS2 沉默逆转了 LPS 对滋养细胞侵袭和 PE 炎症的促进作用,在 PE 分子机制研究方面取得了突破。