Cardiac thromboxane A2 receptor activation does not directly induce cardiomyocyte hypertrophy but does cause cell death that is prevented with gentamicin and 2-APB.

BACKGROUND: We have previously shown that the thromboxane (TXA2) receptor agonist, U46619, can directly induce ventricular arrhythmias that were associated with increases in intracellular calcium in cardiomyocytes. Since TXA2 is an inflammatory mediator and induces direct calcium changes in cardiomyocytes, we hypothesized that TXA2 released during ischemia or inflammation could also cause cardiac remodeling. METHODS: U46619 (0.1-10 µM) was applied to isolated adult mouse ventricular primary cardiomyocytes, mouse ventricular cardiac muscle strips, and cultured HL-1 cardiomyocytes and markers of hypertrophy and cell death were measured. RESULTS: We found that TXA2 receptors were expressed in ventricular cardiomyocytes and were functional via calcium imaging. U46619 treatment for 24 h did not increase expression of pathological hypertrophy genes (atrial natriuretic peptide, ß-myosin heavy chain, skeletal muscle α-actin) and it did not increase protein synthesis. There was also no increase in cardiomyocyte size after 48 h treatment with U46619 as measured by flow cytometry. However, U46619 (0.1-10 µM) caused a concentration-dependent increase in cardiomyocyte death (trypan blue, MTT assays, visual cell counts and TUNEL stain) after 24 h. Treatment of cells with the TXA2 receptor antagonist SQ29548 and inhibitors of the IP3 pathway, gentamicin and 2-APB, eliminated the increase in cell death induced by U46619. CONCLUSIONS: Our data suggests that TXA2 does not induce cardiac hypertrophy, but does induce cell death that is mediated in part by IP3 signaling pathways. These findings may provide important therapeutic targets for inflammatory-induced cardiac apoptosis that can lead to heart failure.

FGF23 is a novel regulator of intracellular calcium and cardiac contractility in addition to cardiac hypertrophy.

Fibroblast growth factor 23 (FGF23) is a hormone released primarily by osteocytes that regulates phosphate and vitamin D metabolism. Recent observational studies in humans suggest that circulating FGF23 is independently associated with cardiac hypertrophy and increased mortality, but it is unknown whether FGF23 can directly alter cardiac function. We found that FGF23 significantly increased cardiomyocyte cell size in vitro, the expression of gene markers of cardiac hypertrophy, and total protein content of cardiac muscle. In addition, FGFR1 and FGFR3 mRNA were the most abundantly expressed FGF receptors in cardiomyocytes, and the coreceptor α-klotho was expressed at very low levels. We tested an animal model of chronic kidney disease (Col4a3(-/-) mice) that has elevated serum FGF23. We found elevations in common hypertrophy gene markers in Col4a3(-/-) hearts compared with wild type but did not observe changes in wall thickness or cell size by week 10. However, the Col4a3(-/-) hearts did show reduced fractional shortening (-17%) and ejection fraction (-11%). Acute exposure of primary cardiomyocytes to FGF23 resulted in elevated intracellular Ca(2+) ([Ca(2+)](i); F/F(o) + 86%) which was blocked by verapamil pretreatment. FGF23 also increased ventricular muscle strip contractility (67%), which was inhibited by FGF receptor antagonism. We hypothesize that although FGF23 can acutely increase [Ca(2+)](i), chronically this may lead to decreases in contractile function or stimulate cardiac hypertrophy, as observed with other stress hormones. In conclusion, FGF23 is a novel bone/heart endocrine factor and may be an important mediator of cardiac Ca(2+) regulation and contractile function during chronic kidney disease.

Genotoxicity assessment of oxirane-based dental monomers in mammalian cells.

The potential use of oxirane (epoxy) monomers in dental composite development raises the concern to test their genetic safety. Oxiranes can interact with DNA resulting in DNA damage, mutations, and possibly carcinogenesis. Our objective was to evaluate DNA damage and cell-cycle disruption in mammalian cells after exposure to epoxy monomers. The experimental oxiranes were Araldite trade mark GY 281, Cyracure trade mark UVR 6105 and 1,3-dioxane-2,2'-1,3-dioxane-5',4'-bicyclo[4.1.0] heptane (DECHE-TOSU). L929 fibroblast cells were incubated with the monomer for 7 and 24 h at 37 degrees C/5% CO(2). After incubation, cells were subjected to DNA damage alkaline unwinding assay and flow cytometry cell-cycle analysis. Lack of DNA damage and cell-cycle effects were observed with DECHE-TOSU. Exposure to subtoxic doses of Araldite trade mark GY 281 or Cyracure trade mark UVR 6105 caused DNA damage and cell cycle disruption. A significant (p < 0.01) effect for Araldite trade mark GY 281 was observed with cell populations in G1 and G2/M when compared to DMSO solvent control. Similar comparisons revealed significant differences in G2/M cell cycle population after 24-h exposure to 100 microM Cyracure trade mark UVR 6105. For comparison, BISGMA was evaluated to produce DNA damage but without cell-cycle effects suggesting DNA repair mechanisms were effective. Our findings with DECHE-TOSU, Araldite trade mark GY 281 and Cyracure trade mark UVR 6105 indicated cell-cycle disruption followed DNA damage.