Zhao Ying-Jie, Zhang Si-Yan, Wei Ying-Ying, Li Hui-Hui, Lei Wei, Wang Kai, Kumar Sathish, Zhou Chi, Zheng Jing
Department of Obstetrics and Gynecology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Department of Rheumatology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
Am J Physiol Cell Physiol. 2025 Mar 1;328(3):C954-C966. doi: 10.1152/ajpcell.00849.2024. Epub 2025 Feb 5.
We have reported that an endogenous aryl hydrocarbon receptor (AhR) ligand, 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE), inhibits functions of human umbilical vein endothelial cells (HUVECs) and induces preeclampsia (PE)-like symptoms in rats. Herein, we tested the hypothesis that ITE impairs endothelial functions via disturbing transcriptome and phosphoproteome in HUVECs. We measured AhR activity in human maternal and umbilical vein sera from PE and normotensive (NT) pregnancies. The serum-induced changes in CYP1A1/B1 mRNA (indexes of AhR activation) in HUVECs were quantified using quantitative reverse transcription polymerase chain reaction (RT-qPCR). ITE's effects on endothelial proliferation and monolayer integrity in female and male HUVECs were determined. We profiled ITE-induced changes in transcriptome and phosphoproteome in HUVECs using RNA-seq and bottom-up phosphoproteomics, respectively. After 12 h of treatment, umbilical vein sera from PE increased CYP1A1 mRNA (1.7-fold of NT) in HUVECs, which was blocked by CH223191, an AhR antagonist. ITE dose-dependently inhibited endothelial proliferation (76%-87% of control) and time-dependently reduced endothelial integrity with a maximum inhibition (∼10%) at 40 h. ITE induced 140 and 80 differentially expressed genes in female and male HUVECs, respectively. ITE altered phosphorylation of 92 and 105 proteins at 4 and 24 h, respectively, in HUVECs. These ITE-dysregulated genes and phosphoproteins were enriched in biological functions and pathways that are relevant to heart, liver, and kidney diseases, vascular functions, and inflammatory responses. Thus, endogenous AhR ligands may impair endothelial functions by disturbing transcriptome and phosphoproteome. These AhR ligand-dysregulated genes and phosphoproteins may be therapeutic and cell sex-specific targets for PE-induced endothelial dysfunction. Preeclampsia elevates AhR agonistic activities in fetal circulation and alters immune cell gene signatures of human umbilical vein endothelial cells (HUVECs). An endogenous AhR ligand (ITE) decreases cell proliferation and monolayer integrity in HUVECs in vitro. ITE dysregulates transcriptome in HUVECs in a fetal sex-specific manner. ITE also disrupts phosphoproteome in HUVECs. These ITE-dysregulated genes and phosphoproteins are highly relevant to diseases of the heart, vascular function, and inflammatory responses.
我们曾报道，一种内源性芳烃受体（AhR）配体，2-（1'H-吲哚-3'-羰基）-噻唑-4-羧酸甲酯（ITE），可抑制人脐静脉内皮细胞（HUVECs）的功能，并在大鼠中诱导子痫前期（PE）样症状。在此，我们检验了ITE通过扰乱HUVECs的转录组和磷酸化蛋白质组来损害内皮功能这一假说。我们检测了来自PE和血压正常（NT）妊娠的人类母体和脐静脉血清中的AhR活性。使用定量逆转录聚合酶链反应（RT-qPCR）对HUVECs中血清诱导的CYP1A1/B1 mRNA（AhR激活指标）变化进行定量。测定了ITE对雌性和雄性HUVECs内皮细胞增殖和单层完整性的影响。我们分别使用RNA测序和自下而上的磷酸化蛋白质组学分析了ITE诱导的HUVECs转录组和磷酸化蛋白质组的变化。处理12小时后，PE患者的脐静脉血清使HUVECs中的CYP1A1 mRNA增加（为NT的1.7倍），这被AhR拮抗剂CH223191阻断。ITE剂量依赖性地抑制内皮细胞增殖（为对照的76%-87%），并随时间依赖性地降低内皮完整性，在40小时时抑制作用最大（约10%）。ITE分别在雌性和雄性HUVECs中诱导了140个和80个差异表达基因。ITE分别在4小时和24小时改变了HUVECs中92种和105种蛋白质的磷酸化。这些ITE失调的基因和磷酸化蛋白质在与心脏、肝脏和肾脏疾病、血管功能及炎症反应相关的生物学功能和途径中富集。因此，内源性AhR配体可能通过扰乱转录组和磷酸化蛋白质组来损害内皮功能。这些AhR配体失调的基因和磷酸化蛋白质可能是PE诱导的内皮功能障碍的治疗靶点及细胞性别特异性靶点。子痫前期会提高胎儿循环中的AhR激动活性，并改变人脐静脉内皮细胞（HUVECs）的免疫细胞基因特征。一种内源性AhR配体（ITE）在体外可降低HUVECs的细胞增殖和单层完整性。ITE以胎儿性别特异性方式失调HUVECs的转录组。ITE还会破坏HUVECs的磷酸化蛋白质组。这些ITE失调的基因和磷酸化蛋白质与心脏疾病、血管功能及炎症反应高度相关。
