miR-19a-3p downregulates tissue factor and functions as a potential therapeutic target for sepsis-induced disseminated intravascular coagulation

• Published in: Biochemical pharmacology Vol. 192; p. 114671
• Main Authors: Zhang, Rong, Lu, Sifen, Yang, Xudan, Li, Maojun, Jia, Hui, Liao, Jing, Jing, Qing, Wu, Yanmei, Wang, Haichuan, Xiao, Feng, Bai, Xiaohong, Na, Xiaoxue, Kang, Yulin, Wan, Ling, Yang, Jiyun
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01.10.2021
• Subjects: Biomarker; Coagulation cascade; Disseminated intravascular coagulation; miR-19a-3p; Therapeutic target; Tissue factor; Disseminated intravascular coagulation; Biomarker; miR-19a-3p; Therapeutic target; Tissue factor; Coagulation cascade
• ISSN: 0006-2952; 1873-2968
• DOI: 10.1016/j.bcp.2021.114671

The DIC model is performed in LPS-induced PBMCs and HUVECs. miR-19a-3p can indirectly inhibit the coagulation cascade in DIC by directly inhibiting TF, which is dependent on NF-κB/IκB and AKT pathways. [Display omitted] Sepsis-induced disseminated intravascular coagulation (DIC) is a common life-threatening terminal-stage disease with high mortality. This study aimed to identify effective miRNAs as therapeutic targets for DIC. Bioinformatics and luciferase reporter gene analyses were performed to predict miR-19a-3p and validate that it targets tissue factor (TF). Quantitative real-time PCR was used to detect the expression of miR-19a-3p and TF, and TF procoagulant activity was determined using the chromogenic substrate method. Western blotting was used to detect the protein levels of TF, AKT serine/threonine kinase (AKT), extracellular regulated protein kinases (ERK), nuclear factor kappa B (NF-κB) P65, NFKB inhibitor alpha (IκB-a) and their phosphorylated counterparts in cell experiments. Furthermore, a rat model was established to explore the potential of miR-19a-3p in DIC treatment. As a result, a human clinical study revealed that miR-19a-3p was downregulated and that TF was upregulated in neonates with sepsis-induced DIC compared with those in the control group. The luciferase reporter assay showed that TF was a direct target of miR-19a-3p. Cell experiments verified that the mRNA and protein levels of TF, and the p-AKT/AKT, p-Erk/Erk, p-P65/P65, p-IκB-a/IκB-a ratios, and TF procoagulant activity were significantly decreased in lipopolysaccharide (LPS) -induced human peripheral blood mononuclear cells (PBMCs) and human umbilical vein endothelial cells (HUVECs) inhibited by overexpression of miR-19a-3p, and that miR-19a-3p regulating TF was dependent on the NF-kB and AKT pathways. In vivo, miR-19a-3p injection into DIC rats suppressed the mRNA expression of TF; more importantly, significant improvements in coagulation function indicators and in histopathologies of lung and kidney were observed. In conclusion, miR-19a-3p may suppress DIC by targeting TF and might be a potential therapeutic target in treating sepsis-induced DIC.