Knockdown of lncRNA XIST Ameliorates IL-1β-Induced Apoptosis of HUVECs and Change of Tissue Factor Level via miR-103a-3p/HMGB1 Axis in Deep Venous Thrombosis by Regulating the ROS/NF-κB Signaling Pathway

Abstract

Background. The effect of lncRNA X inactive-specific transcript (XIST) inducing cardiovascular diseases on deep vein thrombosis (DVT) and its mechanism has not been reported. In this study, we uncovered the mystery that lncRNA XIST causes DVT with HUVEC dysfunction. Method. The expression levels of lncRNA XIST and miR-103a-3p were detected by qRT-PCR, and HMGB1 expression was determined by qRT-PCR and western blot. The correlations among the expression levels of lncRNA XIST, miR-103a-3p, and HMGB1 were determined by Spearman’s rank-order correlation test. XIST siRNA (si-XIST) was transfected into HUVECs to knock down the intrinsic expression of lncRNA XIST. The influences of si-XIST on interleukin-1 beta- (IL-1β-) treated HUVEC viability and apoptosis and the level of tissue factor (TF) were detected by MTT, flow cytometry, and ELISA kit, respectively. The relationships between lncRNA XIST, miR-103a-3p, and HMGB1 were predicted by the Encyclopedia of RNA Interactomes (ENCORI) database and verified by dual luciferase reporter assay. The effects of lncRNA XIST and miR-103a-3p on HMGB1 expression were detected by qRT-PCR, western blot, and immunofluorescence analysis. The levels of ROS/NF-κB pathway-related proteins were detected to study the regulatory mechanism of lncRNA XIST/miR-103a-3p/HMGB1 on IL-1β-treated HUVECs apoptosis and change of TF level. Results. The upregulated expression levels of lncRNA XIST and HMGB1 and downregulated level of miR-103a-3p were found in the plasma of DVT patients and IL-1β-treated HUVECs. Si-XIST promoted cell viability and inhibited HUVEC apoptosis and ameliorated the change of TF level triggered by IL-1β. lncRNA XIST sponged miR-103a-3p and miR-103a-3p targeted HMGB1. Si-XIST inhibited the ROS/NF-κB pathway to suppress HUVEC apoptosis and ameliorate the change of TF level induced by IL-1β via the miR-103a-3p/HMGB1 axis. Conclusion. lncRNA XIST sponged miR-103a-3p improving HMGB1 expression to exacerbate DVT by activating the ROS/NF-κB signaling pathway. Our findings indicated that lncRNA XIST can be used as a potential therapeutic target in DVT.

LncRNA NEAT1 Promotes Vascular Endothelial Cell Dysfunction via miR-218-5p/GAB2 and Serves as a Diagnostic Biomarker for Deep Vein Thrombosis

“… 21 Hypertension and diabetes can lead to vascular endothelial cell damage and thrombosis formation due to metabolic disorders. 3 Additionally, NEAT1 has been shown to participate in the alleviation of diabetic retinopathy through crocetin treatment. 22 High glucose induces a time-dependently upregulates of NEAT1 expression, which is elevated in diabetic nephropathy and accelerates proliferation, oxidative stress, and inflammation.…”

“…However, when endothelial cell damage or apoptosis impairscells damage or apoptosis impairs their normal anticoagulant effect, promoting thrombosis, and affecting intravascular reactive substance levels to aggravate thrombosis. 3 , 29 The importance of vascular endothelial cells necessitates investigation into mechanisms in vitro. Mechanistically, we found that NEAT1 significantly inhibited HUVECs proliferation and migration while increasing apoptosis.…”

“…12 Furthermore, atherosclerotic (AS) disease produces lumen constriction and increased blood viscosity, both of which can lead to thrombosis. 3 NEAT1 levels are higher in those with coronary artery disease or carotid AS plaques, and silencing it reduces vascular endothelial cell inflammation. 13 Dou et al discovered that NEAT1 is abnormally high in the serum of individuals with pulmonary hypertension, a typical consequence of DVT.…”

“… 1 DVT, is the world's third most common blood illness, with up to 50 000 new cases per year in the United States and up to 12 million in China. 2 , 3 DVT misdiagnosis is common due to its gradual development and the absence of clinical signs in the early stage. 4 Furthermore, the currently utilized diagnostic biomarker D-dimer is significantly expressed in non-thrombotic illnesses such as stroke, infection, and diffuse vascular coagulation, discounting its specificity.…”

Long Non-Coding RNAs in Venous Thromboembolism: Where Do We Stand?

“…In addition, XIST is also associated with preeclampsia and cancer [100,101]. This lncRNA was found to be highly expressed in the plasma of DVT patients, possibly acting as a decoy by sponging miR-103a-3p, which in turn, targets highmobility group box 1 (HMGB1) [102]. Concordantly, miR-103a-3p expression was found to be reduced in patients with unprovoked VTE [103].…”