Antisense Oligodeoxynucleotide Against Tissue Factor Inhibits Human Umbilical Vein Endothelial Cells Injury Induced by Anoxia-Reoxygenation

Abstract

Aims: The role of antisense oligodeoxynucleotide against tissue factor (aODN/TF) in cultured human umbilical vein endothelial cells (HUVECs) subjected to anoxia-reoxygenation (A/R) was investigated. Methods: HUVECs were divided randomly into control group, A/R group, aODN/TF+A/R group, sense oligodeoxynucleotide (sODN/TF) + A/R group and mismatched oligodeoxynucleotide (mODN/TF) + A/R group, in the latter 3 groups, HUVECs were transfected with aODN/TF, sODN/TF and mODN/TF respectively. HUVECs in all A/R groups underwent 3 hrs of anoxia and followed by 2 hrs of reoxygenation. In order to investigate the potential mechanisms of how increased TF may contribute to A/R injury in HUVECs, another set of HUVECs were incubated with human recombinant active site blocked factor VII (FVIIai) during A/R. Results: After A/R, TF expression at both mRNA and protein level was increased, furthermore, cell viability and the concentrations of SOD, GSH-PX and NO were declined, while LDH, MDA and ET-1 were overproduced (P<0.05 to 0.001 versus control group). In HUVECs of aODN/TF+A/R group, however, TF expression was inhibited, while the declined cell viability and the concentrations of SOD, GSH-PX, NO as well as the enhanced LDH, MDA and ET-1 levels occurred during A/R were ameliorated and reversed effectively (P<0.05 to 0.01 versus those in other A/R groups). The results also showed that ROS was increased and PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase were all activated after A/R (P<0.001 versus HUVECs under normoxia), while FVIIai inhibited the increment of ROS, PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase, and improved the changes of TF:C, MDA, SOD, GSH-PX, cell viability and LDH occurred during A/R (P<0.05 to 0.001 versus HUVECs without FVIIai treatment). Conclusion: Tissue factor plays an important role in the development of HUVECs injury induced by anoxia-reoxygenation, inhibition of TF with antisense oligodeoxynucleotide is an effective approach to ameliorate the damage.

Tissue Factor, Blood Coagulation, and Beyond: An Overview

“…On the molecular biology front, miR-19 [61], short hairpin RNA [62], hairpin ribozyme [63], or antisense ODN [64–66] readily downregulates TF mRNA translation and expression.…”

“…Metformin, an antidiabetic agent, suppresses the production of TNF α [59], a known factor for insulin resistance [213–215]. Antisense oligonucleotide blocking TF expression prevents leukocyte adhesion following renal ischemic reperfusion injury [66, 270]. COX inhibitors readily show anti-inflammation [152, 153] as well as antithrombosis.…”

Tissue factor antisense deoxyoligonucleotide prevents monocrotaline/LPS hepatotoxicity in mice

“…Interestingly, treating rats with TF‐ASO successfully reduced ischemia–reperfusion‐induced liver injury (Nakamura et al ., ). Similarly, in vitro treatment of human umbilical endothelial cells with TF‐ASO diminished injury induced by anoxia–reoxygenation (Yin et al ., ). We developed a hepatotoxicity model in mice by administration of a sub‐toxic p.o.…”

Heparin-binding epidermal growth factor–like growth factor (HB-EGF) preserves gut barrier function by blocking neutrophil-endothelial cell adhesion after hemorrhagic shock and resuscitation in mice

“…10 To examine PMN-EC interactions, an in vitro PMN-EC adhesion model was established. 43 In this model, EC injured by A/R express various inflammatory mediators such as adhesion molecules, interleukins, growth factors, cytokines and chemokines, 44 facilitating PMN-EC adherence. We found that treatment of PMN with HB-EGF significantly decreased PMN-EC adherence 4 h after A/R, and this effect was reversed with EGFR inhibition.…”