Low extracellular calcium enhances beta cell sensitivity to the stimulatory influence of 1,25-dihydroxyvitamin D3 on insulin release by islets from vitamin D3-deficient rats.

The beneficial effect of 1,25-dihydroxyvitamin D3 [1,25 (OH)2 D3] on insulin secretion from beta cells in hypocalcemic vitamin D3-deficient rats is now well established. Moreover, few data concerning the mechanism of 1,25 (OH) 2D3 efficiency as a function of the severity of hypocalcemia. In the present experiment, we submitted islets from vitamin D3-deficient rats to in vitro exposure to a range of decreasing extracellular Ca2+ concentrations ([Ca2+]ex), from 0.5 mM to 0.6 mM, during a 6-h 10-8 M 1,25 (OH) 2D3 induction. Thereafter, we compared the effect of this pretreatment on the islets' insulin response to a given stimulus. Various stimuli were used, and we measured in parallel the variations of 86Rb+ and 45Ca2+ efflux and insulin release into the perifusion medium. In the presence of 1,25 (OH) 2D3, we observed an inverse correlation between the [Ca2+]ex pre-exposure and the amplitude of the insulin response to certain stimuli studied, suggesting that beta cells that were pre-exposed to low [Ca2+]ex became more sensitive to the beneficial effect of 1,25 (OH) 2D3 on insulin release. This effect was observed when beta cells were activated by acetylcholine but only during its second phase of stimulation, and more particularly with the barium plus theophylline stimulus. In contrast, insulin release was not affected by [Ca2+]ex pre-exposure during 1,25 (OH) 2D3 induction in response to acetylcholine during its first phase of stimulation, thus excluding any mechanism mediated via nutrient pathways, membrane depolarization, or inositol triphosphate (IP3)-dependent events. Moreover, the islets that were pre-exposed to a 10-fold [Ca2+]ex exhibited only a 50% lower 45Ca2+ content after 45Ca2+ loading, suggesting a different or relatively more efficient storage capacity in the presence of low extracellular calcium. Studies of 45Ca2+ efflux showed that the mobilization of Ca2+ stores induced by a barium plus theophylline stimulus, in the absence of calcium in the perifusion medium, was more efficient in islets pre-exposed to low [Ca2+]ex, whereas the acetylcholine-IP 3-induced mobilization of Ca2+ from reticular stores was not affected. These results generated the hypothesis that 1,25 (OH)2D3 may prepare the beta cells during their pre-exposure to low [Ca2+]ex to become more efficient as concerns insulin release via a more efficient mobilization of 45Ca2+ stores (mitochondrial?) and by an activation of release potentiating systems via protein kinase C protein kinase A pathways.

Stimulatory effect of 1,25-dihydroxyvitamin D3 on calcium handling and insulin secretion by islets from vitamin D3-deficient rats.

Among the various vitamin D3 metabolites, 1,25-(OH)2D3 is the specific secosteroid hormone that can enhance, in vitro, the weak insulin response to glucose of islets from vitamin D3-deficient rats. Because this potentiating effect is preceded by an increase in Ca2+ handling, several putative sites of action were studied by measuring 45Ca2+ and 86Rb+ (as K+ tracer) efflux during perifusions in the presence of various stimuli known to affect Ca2+ movements in different ways: high glucose without calcium, high calcium without glucose, high potassium, or barium-theophylline without calcium or glucose. The present results show that 1,25-(OH)2D3 may activate Ca2+ handling by at least two mechanisms: (1) an increase of Ca2+ entry via voltage-dependent Ca2+ channels in the experiments in which extracellular Ca2+ was present and where Ca2+ channels were opened; this 1,25-(OH)2D3 influence on Ca2+ channels was not mediated by a possible indirect influence on K+ channels because 86Rb+ fluxes were never observed to be affected by the steroid; (2) an enhancement of 45Ca2+ mobilization from intracellular stores as suggested by barium-theophylline stimulation and probably also via the Ca2+ stimulus. Both of these 1,25-(OH)2D3 influences tended to provide more calcium to the B cell of vitamin D3-deficient rats. But this prerequisite was not sufficient in itself to potentiate the insulin response; indeed, experiments with barium-theophylline suggested that 1,25-(OH)2D3 may also activate a cAMP-dependent exocytosis process.

Renal cytotoxicity of cisplatin in cultured glomerular mesangial and proximal and distal tubular cells.

Cisplatin is of considerable therapeutic value owing to its anti-tumoral activity. Unfortunately, its nephrotoxicity can reduce its clinical use. In vivo toxicity studies have shown large renal haemodynamic changes and differential tubular nephrotoxicity with strong proximal tropism. The present study compared renal cytotoxicity of cisplatin in three different cell cultures: glomerular mesangial cells and two renal tubular cell lines, LLCPK(1) (proximal) and MDCK (distal). Cell viability was assessed by the neutral red test. Cisplatin at 10(-4)m induced a similar cell mortality in the three targets (about 80%). Mesangial cell mortality was concentration dependent, at 82, 31, 19 and 12% for 10(-4)-10(-7)m, respectively. The IC(50) for MDCK cells was 5.35 x 10(-5)m compared with 3.25 x 10(-5)m for LLCPK(1). For the different cisplatin concentrations mortality was two to three times higher in LLCPK(1), confirming the strong proximal tropism of cisplatin. These results demonstrate cisplatin cytotoxicity at both the glomerular and tubular levels, and open the way for comparative studies with new cisplatin derivatives for the optimization of their clinical use.