Thymoquinone Modulates Blood Coagulation in Vitro via Its Effects on Inflammatory and Coagulation Pathways

Abstract

Thymoquinone (THQ) is a major component of black seeds. Given that both THQ and black seeds exhibit anti-cancer and anti-inflammatory activities, we hypothesized that THQ will affect cancer-associated thrombosis (CAT), which is primarily triggered by tissue factor (TF) and inflammation. The effect of both black seed-extracted and purchased (“pure”) THQ on normal blood coagulation was tested with in vitro thromboelastography (TEG) and activated partial thromboplastin time (aPTT) coagulation assays. The effect of pure THQ on CAT was tested with aPTT assay using pancreatic cancer cell lines that are either positive or negative for TF, and with TEG assay using lipopolysaccharide as an inflammatory trigger. Additionally, the direct effect of THQ on the inactivation of factors IIa and Xa was assessed. Since TNF-α facilitates crosstalk between inflammation and thrombosis by triggering the NF-κB pathway, we tested THQ’s ability to interfere with this communication with a luciferase assay. Both extracted and pure THQ had minimal effects on normal blood coagulation. Pure THQ reversed CAT initiated by both TF and inflammation to basal levels (p < 0.001). Mechanistically, while THQ had minimal to no effect on factor IIa and Xa inactivation, it strongly reduced the effects of TNF-α on NF-κB elements (p < 0.001). THQ has a minimal effect on basal coagulation and can reverse CAT in vitro, possibly by interfering with the crosstalk between inflammation and coagulation. This study suggests the utility of THQ as a preventative anticoagulant and/or as a supplement to existing chemotherapies and anticoagulant therapies.

Prophetic Medicine-Nigella Sativa (Black cumin seeds) – Potential herb for COVID-19?

“…In vitro coagulation assays of thromboelastography (TEG) and activated partial thromboplastin time (aPTT) using pancreatic cancer cell lines, tested the anticoagulant effect of Thymoquinone of N. sativa and it has been determined that Thymoquinone reversed the cancer-associated thrombosis (CAT) by returning tissue factor (TF) and inflammation to basal levels. Moreover, Thymoquinone inhibited the activity of factor Xa significantly, at higher concentrations (0.8-1.6 mg/mL) [ 52 ].…”

In vitro Evaluation of the Anti-inflammatory Effects of Thymoquinone in Osteoarthritis and in silico Analysis of Inter-Related Pathways in Age-Related Degenerative Diseases

“…It was evident that TQ at higher concentrations (5 μM) led to morphological changes and cell death compared to lower concentrations. In the present study, we identified that the IC50 concentration of TQ to be 3 μM based on the mean concentrations from two different assays (MTT and CellTiter-Blue ® ) at 48 h. The concentrations of TQ and their cellular effects varied from 0.1 to 100 μM based on the purity of the compound, the cell types used and the nature of the studies ( Muralidharan-Chari et al, 2016 ; Aziz et al, 2018 ; Bordoni et al, 2019 ).…”

Thymoquinone: A Promising Natural Compound with Potential Benefits for COVID-19 Prevention and Cure

“…N sativa , due to its wide range of bioactive components such as TQ and nigellimine, could offer a range of benefits for treating COVID-19, such as blocking the introduction of the virus to pneumocytes; providing ionophores to improve zinc intake, thereby improving the host immune response to SARS-CoV-2; and preventing the virus from replicating. 29 TQ is the main bioactive principle in N Sativa , and this has been found to confer a range of therapeutic advantages 34 including antioxidant, 38 anti-inflammatory, 39 , 46 anticancer, 40 antibacterial, 41 antifungal, 42 anticoagulant, 47 anti-sepsis, 48 and anticonvulsant activity. 43 N Sativa seeds have also demonstrated immunomodulatory effects, 49 , 50 while several studies suggest that N Sativa seeds have some antiviral effects.…”