Li Jiang, Huang Hong, Xu Shanhu, Fan Mengge, Wang Kaili, Wang Xia, Zhang Jiao, Huang Shengshi, Gatt Alex, Liu Ju
Medical Research Center, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, Shandong 250014, P.R. China.
Institute of Microvascular Medicine, Medical Research Center, The First Affiliated Hospital of Shandong First Medical University and Shandong Provincial Qianfoshan Hospital, Jinan, Shandong 250014, P.R. China.
Exp Ther Med. 2023 Jul 10;26(2):408. doi: 10.3892/etm.2023.12107. eCollection 2023 Aug.
Complement factor H (CFH), a major soluble inhibitor of complement, is a plasma protein that directly interacts with the endothelium of blood vessels. Mutations in the CFH gene lead to diseases associated with excessive angiogenesis; however, the underlying mechanisms are unknown. The present study aimed to determine the effects of CFH on endothelial cells and to explore the underlying mechanisms. The adenoviral plasmid expressing CFH was transduced into HepG2 cells, and the culture medium supernatant was collected and co-cultured with human umbilical vein endothelial cells (HUVECs). Cell proliferation was measured by CCK8 and MTT assays, and cell migration was measured by wound healing and Transwell assays. Reverse transcription-quantitative PCR was performed to detect gene transcription. Western blotting was used to determine protein levels. The results revealed that CFH can inhibit migration, but not viability, of HUVECs. In addition, CFH did not significantly alter MAPK or TGF-β signaling, whereas it decreased STAT3 phosphorylation in HUVECs. Furthermore, CFH failed to reduce migration of HUVECs, with inhibition of STAT3 signaling by STATTIC or activation of STAT3 signaling by overexpression of STAT3 (Y705D) compromising CFH-inhibited HUVEC migration. CFH also decreased the expression levels of vascular endothelial growth factor receptor 2, a downstream effector of STAT3 mediating endothelial cell migration. In conclusion, the present study suggested that CFH may be a potential therapeutic target for angiogenesis-related diseases.
补体因子H(CFH)是补体的主要可溶性抑制剂,是一种直接与血管内皮相互作用的血浆蛋白。CFH基因突变会导致与血管生成过度相关的疾病;然而,其潜在机制尚不清楚。本研究旨在确定CFH对内皮细胞的影响,并探索其潜在机制。将表达CFH的腺病毒质粒转导至HepG2细胞中,收集培养基上清液并与人脐静脉内皮细胞(HUVECs)共培养。通过CCK8和MTT法检测细胞增殖,通过伤口愈合和Transwell法检测细胞迁移。进行逆转录定量PCR以检测基因转录。使用蛋白质印迹法测定蛋白质水平。结果显示,CFH可抑制HUVECs的迁移,但不影响其活力。此外,CFH对MAPK或TGF-β信号通路无显著影响,而它可降低HUVECs中STAT3的磷酸化水平。此外,CFH未能降低HUVECs的迁移,用STATTIC抑制STAT3信号通路或通过过表达STAT3(Y705D)激活STAT3信号通路均会损害CFH对HUVEC迁移的抑制作用。CFH还降低了血管内皮生长因子受体2的表达水平,血管内皮生长因子受体2是STAT3介导内皮细胞迁移的下游效应分子。总之,本研究表明CFH可能是血管生成相关疾病的潜在治疗靶点。