He Junbing, Shao Zixuan, Li Zhuoji, He Yufu, Zhang Jingqi, Zhong Haotian, Li Jiekai, Liu Qinghua, Shao Yiming
The Department of Emergency, The First Affiliated Hospital, Jinan University, Guangzhou, GD, China.
Jieyang Medical Research Center, Jieyang People's Hospital, Jieyang Affiliated Hospital of Sun Yat-sen University, Jieyang, GD, China.
Immunol Invest. 2025 Aug;54(6):867-893. doi: 10.1080/08820139.2025.2503174. Epub 2025 May 14.
HOX transcript antisense intergenic RNA (HOTAIR) has been implicated in inflammation and vascular pathology, but its role in regulation of ADAM17 and sepsis-induced endothelial injury remains unclear.
LPS-treated human umbilical vein endothelial cells (HUVECs) modeled sepsis-induced endothelial injury, which were assessed via qRT-PCR, western blot and immunofluorescence. HOTAIR-knockout mice were treated with cecal ligation and perforation to establish sepsis model.
LPS-stimulation increased expression of HOTAIR and ADAM17 and decreased miR-326 levels in HUVECs. HOTAIR-knockdown by antisense oligonucleotides (ASOs) decreased ADAM17, TNF-α production and NF-κB activities; it also alleviated endothelial inflammation, VE-cadherin integrity damage, apoptosis and barrier dysfunction, while miR-326 inhibition reversed these effects. MiR-326 inhibited TNF-α/NF-κB via targeting ADAM17. Further experiments demonstrated recombinant TNF-α reversed the inhibitory effect of HOTAIR-ASOs on LPS-triggered TNF-α/NF-κB activation and downstream endothelial injury, which were further mitigated by NF-κB or p38 MAPK inhibitors. In-vivo experiments in HOTAIR-knockout mice confirmed the role of HOTAIR/miR-326/ADAM17 in regulating NF-κB and p38 MAPK inflammation, with improved lung injury and survival following sepsis.
The HOTAIR/miR-326/ADAM17 axis is a key regulator of inflammation, endothelial injury and barrier dysfunction during sepsis via modulation of TNF-α/NF-κB signaling, providing new insights into the mechanisms underlying endothelial injury in sepsis.
HOX转录反义基因间RNA（HOTAIR）与炎症和血管病变有关，但其在调节ADAM17和脓毒症诱导的内皮损伤中的作用尚不清楚。
用脂多糖（LPS）处理人脐静脉内皮细胞（HUVECs）建立脓毒症诱导的内皮损伤模型，通过定量逆转录聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法和免疫荧光法进行评估。用盲肠结扎穿孔术处理HOTAIR基因敲除小鼠以建立脓毒症模型。
LPS刺激增加了HUVECs中HOTAIR和ADAM17的表达，并降低了miR-326水平。反义寡核苷酸（ASOs）敲低HOTAIR可降低ADAM17、肿瘤坏死因子-α（TNF-α）的产生和核因子-κB（NF-κB）活性；还可减轻内皮炎症、血管内皮钙黏蛋白完整性损伤、细胞凋亡和屏障功能障碍，而抑制miR-326可逆转这些作用。miR-326通过靶向ADAM17抑制TNF-α/NF-κB。进一步实验表明，重组TNF-α可逆转HOTAIR-ASOs对LPS触发的TNF-α/NF-κB激活及下游内皮损伤的抑制作用，而NF-κB或p38丝裂原活化蛋白激酶（MAPK）抑制剂可进一步减轻这种损伤。对HOTAIR基因敲除小鼠进行的体内实验证实了HOTAIR/miR-326/ADAM17在调节NF-κB和p38 MAPK炎症中的作用，脓毒症后肺损伤减轻，生存率提高。
HOTAIR/miR-326/ADAM17轴是脓毒症期间炎症、内皮损伤和屏障功能障碍的关键调节因子，通过调节TNF-α/NF-κB信号通路，为脓毒症内皮损伤的潜在机制提供了新见解。
