beraprost and ioversol

We reported previously that radiocontrast medium induces caspase-dependent apoptosis and that cAMP analogs inhibit cell injury in cultured renal tubular cells. In the present study, cellular mechanisms underlying the protective effects of cAMP were determined. Ioversol, a radiocontrast medium, caused cell injury accompanied by decreases in Bcl-2, increases in Bax, and caspase activation in LLC-PK1 cells. Both cell injury and cellular events induced by ioversol were inhibited by dibutyryl cAMP and the prostacyclin analog beraprost. Dibutyryl cAMP increased phosphorylation of Akt and CREB, both of which were reversed by H89, wortmannin and the Akt inhibitor SH-6. The protective effect of dibutyryl cAMP was also reversed by these kinase inhibitors. In dominant-negative CREB-transfected cells, dibutyryl cAMP no longer prevented cell injury or inhibited changes in mRNA expression of Bcl-2 and Bax. In mice with unilateral renal occlusion, ioversol increased urinary excretion of N-acetyl-beta-d-glucosaminidase with concomitant decreases in Bcl-2 mRNA, increases in Bax mRNA, activation of caspase-3, and induction of apoptosis in tubular and interstitial cells. Beraprost completely reversed these in vivo effects of ioversol. These findings suggest that elevation of endogenous cAMP effectively prevents radiocontrast nephropathy through activation of A kinase/PI 3-kinase/Akt followed by CREB phosphorylation and enhanced expression of Bcl-2.

Topics: Animals; Apoptosis; Bucladesine; Caspases; Cell Survival; Contrast Media; Cyclic AMP Response Element-Binding Protein; Enzyme Activation; Epoprostenol; Kidney; Kidney Diseases; LLC-PK1 Cells; Mice; Phosphatidylinositol 3-Kinases; Phosphorylation; Phosphotransferases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Swine; Triiodobenzoic Acids

We previously reported that the apoptotic injury in a porcine renal tubular cell line LLC-PK1 cells induced by radiographic contrast media is attenuated by dibutyl cyclic adenosine monophosphate (cAMP) in a manner dependent on protein kinase A (PKA). The present study was designed to determine whether the elevation of endogenous cAMP with beraprost sodium, a prostacyclin analog, reduces the contrast material-induced renal tubular injury.. The cell injury was induced by the exposure to ioversol for 30 minutes followed by further incubation for 24 hours in the absence of the contrast medium, and assessed by propidium iodide uptake and WST-8 assay. Apoptosis was determined by annexin V stain and DNA electrophoresis. Caspase activity was assessed by the enzymatic degradation of specific substrate peptides. Bax and bcl-2 mRNA expression were determined by reverse transcription-polymerase chain reaction (RT-PCR). The phosphorylation of cAMP-responsive element binding protein (CREB) was measured by an immunofluorescent method.. Beraprost sodium (10 to 1000 nmol/L) attenuated concentration dependently the ioversol-induced decrease in cell viability, in which the protective effect of beraprost sodium was dependent on the elevation of cellular cAMP content. The phosphorylation of CREB was enhanced by beraprost sodium in PKA-dependent manner. In addition, beraprost sodium reversed the ioversol-induced increase in bax mRNA with a concomitant decrease in bcl-2 mRNA and subsequent activation of caspase-3 and -9, thereby resulting in the inhibition of the nuclear damage.. Beraprost sodium reversed the contrast media-induced renal tubular cells in culture by activating cAMP/protein kinase A-dependent phosphorylation of CREB and subsequent enhancement of bcl-2 expression.

Topics: Animals; Apoptosis; Base Sequence; bcl-2-Associated X Protein; Caspases; Contrast Media; Cyclic AMP Response Element-Binding Protein; Enzyme Activation; Epoprostenol; Gene Expression; Genes, bcl-2; Kidney Tubules; LLC-PK1 Cells; Phosphorylation; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; Swine; Triiodobenzoic Acids