Role of HERC5 silencing in modulating apoptosis of EA.hy926 ‎endothelial cells: A Preliminary in vitro study with implications for deep ‎vein thrombosis

Yiming Su; Pimporn Thongmuang; Sarisak Soontornchai; Guangqiu Qin

ผู้แต่ง

Yiming Su Student of Doctoral Degree in Public Health Program, Graduate School, Suan Sunandha Rajabhat University
Pimporn Thongmuang Public Health Program, Graduate School, Suan Sunandha Rajabhat University
Sarisak Soontornchai Public Health Program, Graduate School, Suan Sunandha Rajabhat University
Guangqiu Qin School of Public Health and Management, Guangxi University of Chinese Medicine, Guangxi, China

คำสำคัญ:

HERC5, Vascular endothelial cells, Apoptosis, EA.hy926, Deep vein thrombosis

บทคัดย่อ

Endothelial cell apoptosis contributes to a variety of vascular pathologies, including endothelial dysfunction implicated in deep vein thrombosis (DVT). HERC5, a HECT-domain E3 ubiquitin ligase tightly regulated in activated endothelial cells, has been reported to participate in inflammatory and immune responses, but its role in endothelial cell apoptosis remains insufficiently characterized. This preliminary in vitro study aimed to investigate the effect of HERC5 silencing on apoptosis of EA.hy926 endothelial cells. Three short hairpin RNAs targeting HERC5 (shHERC5_1, shHERC5_2, and shHERC5_3) were delivered into EA.hy926 cells by lentiviral transduction; non-targeting shRNA (shNC) and untransduced cells (Mock) served as controls. Silencing efficiency was confirmed by RT-qPCR and Western blot. Apoptosis was assessed by Annexin V/Propidium Iodide flow cytometry using shHERC5_3, which produced the greatest protein-level knockdown. Compared with shNC, all three shHERC5 constructs significantly reduced HERC5 mRNA and protein levels (P < 0.05). Flow cytometry showed that HERC5 silencing increased the proportion of late apoptotic cells and decreased the proportion of viable cells relative to shNC. These results provide preliminary in vitro evidence that HERC5 may influence endothelial cell apoptosis. Because endothelial apoptosis is one of several mechanisms associated with endothelial dysfunction and vascular pathologies including DVT, further investigation incorporating thrombosis-related endpoints and in vivo models is warranted to clarify whether HERC5 is mechanistically linked to DVT pathogenesis.

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