ADAMTS13 inhibits H2O2-induced human venous endothelial cell injury to attenuate deep-vein thrombosis by blocking the p38/ERK signaling pathway

Guangfeng Zheng; Qiang Zhang; Chuanyong Li; Weijian Fan; Zhichang Pan; Yuting Zhou; Yan Chen; Jianjie Rong

doi:10.4103/cjop.CJOP-D-23-00101

ADAMTS13 inhibits H₂O₂-induced human venous endothelial cell injury to attenuate deep-vein thrombosis by blocking the p38/ERK signaling pathway

Chin J Physiol. 2023 Nov-Dec;66(6):466-473. doi: 10.4103/cjop.CJOP-D-23-00101.

Authors

Guangfeng Zheng¹, Qiang Zhang¹, Chuanyong Li¹, Weijian Fan¹, Zhichang Pan¹, Yuting Zhou², Yan Chen², Jianjie Rong¹

Affiliations

¹ Department of Vascular Surgery, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou, Jiangsu, China.
² Department of Operating Room, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou, Jiangsu, China.

PMID: 38149559
DOI: 10.4103/cjop.CJOP-D-23-00101

Abstract

Deep vein thrombosis (DVT) is a common complication in hematologic malignancies and immunologic disorders. Endothelial cell injury and dysfunction comprise the critical contributor for the development of DVT. A disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13), a plasma metalloprotease that cleaves von Willebrand factor, acts as a critical regulator in normal hemostasis. This study was aimed to explore the role of ADAMTS13 in endothelial cell injury during DVT and the possible mechanism. First, human umbilical vein endothelial cells (HUVECs) were exposed to hydrogen peroxide (H₂O₂). Then, the mRNA and protein expressions of ADAMTS13 were evaluated with the reverse transcription-quantitative polymerase chain reaction and western blot. After treatment with recombinant ADAMTS13 (rADAMTS13; rA13), the viability and apoptosis of H₂O₂-induced HUVECs were assessed by cell counting kit-8 assay and terminal-deoxynucleoitidyl transferase-mediated nick end labeling staining. In addition, the levels of prostaglandin F1-alpha, endothelin-1, and reactive oxygen species were detected using the enzyme-linked immunosorbent assay and dichloro-dihydro-fluorescein diacetate assay. The expressions of proteins related to p38/extracellular signal-regulated kinase (ERK) signaling pathway were estimated with the western blot. Then, p79350 (p38 agonist) was used to pretreat cells to analyze the regulatory effects of rA13 on p38/ERK signaling in H₂O₂-induced HUVEC injury. The results revealed that ADAMTS13 expression was significantly downregulated in H₂O₂-induced HUVECs. The reduced viability and increased apoptosis of HUVECs induced by H₂O₂ were revived by ADAMTS13. ADAMTS13 also suppressed the oxidative stress in HUVECs after H₂O₂ treatment. Besides, ADAMTS13 was found to block p38/ERK signaling pathway, and p79350 reversed the impacts of ADAMTS13 on the damage of HUVECs induced by H₂O₂. To sum up, ADAMTS13 could alleviate H₂O₂-induced HUVEC injury through the inhibition of p38/ERK signaling pathway.

Keywords: ADAMTS13; apoptosis; deep vein thrombosis; oxidative stress; p38/ERK signaling pathway.

MeSH terms

ADAMTS13 Protein* / metabolism
Human Umbilical Vein Endothelial Cells
Humans
Hydrogen Peroxide / adverse effects
MAP Kinase Signaling System*
Oxidative Stress
Venous Thrombosis* / metabolism

Substances

ADAMTS13 protein, human
Hydrogen Peroxide
ADAMTS13 Protein