Cellular and molecular studies of the effects of a selective COX-2 inhibitor celecoxib in the cardiac cell line H9c2 and their correlation with death mechanisms

Abstract

Cardiovascular disease is one of the leading causes of death worldwide, and evidence indicates a correlation between the inflammatory process and cardiac dysfunction. Selective inhibitors of cyclooxygenase-2 (COX-2) enzyme are not recommended for long-term use because of potentially severe side effects to the heart. Considering this and the frequent prescribing of commercial celecoxib, the present study analyzed cellular and molecular effects of 1 and 10 µM celecoxib in a cell culture model. After a 24-h incubation, celecoxib reduced cell viability in a dose-dependent manner as also demonstrated in MTT assays. Furthermore, reverse transcription-polymerase chain reaction analysis showed that the drug modulated the expression level of genes related to death pathways, and Western blot analyses demonstrated a modulatory effect of the drug on COX-2 protein levels in cardiac cells. In addition, the results demonstrated a downregulation of prostaglandin E2 production by the cardiac cells incubated with celecoxib, in a dose-specific manner. These results are consistent with the decrease in cell viability and the presence of necrotic processes shown by Fourier transform infrared analysis, suggesting a direct correlation of prostanoids in cellular homeostasis and survival.

NSAIDs and Cardiovascular Diseases: Role of Reactive Oxygen Species

“…Rat embryonic H9c2 cardiac cells exposed to celecoxib, at concentrations of 10 μ M and 100 μ M, demonstrated a decrease in cell viability by 45% and 92%, respectively, and this was associated with a decrease in the expression levels of the cell survival protein Bcl2 [ 146 ]. However, 1 μ M of celecoxib had no significant effect on cell viability and was associated with an upregulation of the Bcl 2 transcript level by ~30%.…”

“…This effect of such a low amount of NSAID in the upregulation of cell survival gene expression is rather comparable to the effect of proteasome inhibitors like MG-132 or epoxomicin which at nanomolar ranges has been reported to actually increase the proteasome activity in the pretreated neocortical neurons [ 147 ]. Also, this was the first study that measured COX-2 levels directly in the cardiac cells [ 146 ]. Although no significant changes in the COX-2 levels were observed in cells treated with 1 μ M celecoxib, at 10 μ M the COX-2 levels decreased [ 146 ].…”

“…Also, this was the first study that measured COX-2 levels directly in the cardiac cells [ 146 ]. Although no significant changes in the COX-2 levels were observed in cells treated with 1 μ M celecoxib, at 10 μ M the COX-2 levels decreased [ 146 ].…”

Role of cyclooxygenase-2 in Trypanosoma cruzisurvival in the early stages of parasite host-cell interaction

“…PGE2 levels in T. cruzi -infected cells were down-regulated at all investigated time-points compared with the levels in uninfected cultures (145.83 pg/mL). This prostanoid was chosen based on the relevance of PGE2 to the parasite infection ( Sterin-Borda et al 1996 , Abdalla et al 2008 ) and by the fact that H9c2 responds to a different set of stimuli similar to an adult cardiomyocyte ( Kimes & Brandt 1976 ); in addition, there is evidence that describes the production of PGE2 by such cells, when COX-2 is activated ( Lu & Choy 2004 , Kwak et al 2010 , Sharma et al 2013 , Sakane et al 2014) . In the analysed time-points (control, 2, 6, 12, 24 and 48 h), PGE2 levels oscillated and were reduced when compared with the control (p < 0.001): 38.08 pg/mL (2 h), 66.26 pg/mL (6 h), 47.31 pg/mL (12 h), 39.11 pg/mL (24 h) and 52.64 pg/mL (48 h).…”

Cellular and molecular effects of Baccharis dracunculifolia D.C. and Plectranthus barbatus Andrews medicinal plant extracts on retinoid metabolism in the human hepatic stellate cell LX-2

“…Fluorescence microscopy analyses were performed adapting the protocol described in da Silva et al (2016) [35]. For that, 3.4 × 10 4 cells/ well were seeded in 24-well plates containing circular-coverslips and treated as described, previously.…”