Investigating the mechanism by which Naproxen and Prometryn causes Cardiovascular dysfunction

Abstract

Background and ObjectiveCardiovascular disease (CVD) is the leading cause of death and a major cause of disability worldwide. Previous studies have shown that the long‐term and chronic use of nonsteroidal anti‐inflammatory drugs (NSAIDs) increases the risk of developing stroke and cardiovascular diseases. Naproxen is an FDA‐approved NSAID that is currently used to treat several medical conditions such as pain, osteoarthritis, rheumatoid arthritis, polyarticular juvenile idiopathic arthritis, tendonitis, etc. Acetaminophen, the active ingredient in Tylenol, is not an NSAID but it has been associated with increased risk of liver toxicity. Similarly, the long‐term exposures to pesticides such as Prometryn amongst others have been established to be dangerous to human lives and capable to disrupt the physiology of different organs in the body, including the cardiovascular system. Prometryn, a diamino‐1,3,5‐triazine, is used as a selective pre‐ and post‐emergence herbicide. It is used to control annual grasses and broadleaf weeds in cotton and celery. This study aimed to investigate the effects of Naproxen, Acetaminophen, and Prometryn in cardiomyocytes and the underlying mechanisms by which it causes cardiovascular dysfunction.MethodsThe embryonic cardiomyocyte cell line H9c2 is commonly used in numerous in vitro studies because morphological parameters of their cells resemble immature embryonic cardiomyocytes. It has been previously shown that H9c2 cell line and primary neonatal cardiomyocyte cells show similar hypertrophic responses in vitro. H9c2 cardiomyocytes were plated in a 96 well plate at a concentration of 4000 cells/well. The cells were then treated with different concentrations of Naproxen, Acetaminophen, and Prometryn for 24h at 37°C and 5% CO2. The cell viability, reactive oxygen species (ROS) formation, and mitochondrial membrane potential (ΔΨm) were examined.ResultsH9c2 cells treated with 100μM Acetaminophen and 20μM Prometryn showed significant decreases (> 30%) in cell viability. Hydrogen peroxide (H2O2), which was used as a positive control, also resulted in significant decreases in cell viability at 100μM. ROS formation was significantly increased in the presence of 200–400μM Naproxen. Acetaminophen showed significant amount of ROS formation at a higher concentration of 200μM while Prometryn also showed a significant increase in ROS formation at a higher concentration of 30μM. The effect of varying concentrations of Naproxen showed that ΔΨm was decreased by ~30% at 100μM. Prometryn decreased ΔΨm by ~20% at 10–20μM while Acetaminophen also decreased ΔΨm by ~20% at 100–200μM.ConclusionThese results suggest that both concentration and exposure time are important for NSAID‐induced cardiotoxicity. Acetaminophen, which is not an NSAID, increased ROS generation and decreased ΔΨm at higher concentration. This experimental data suggests that Naproxen, Acetaminophen, and Prometryn causes cardiomyocyte dysfunction at varying concentrations.