Pre-clinical methods for the determination of insulin sensitivity.

We compared the hyperinsulinaemic euglycaemic glucose clamping (HEGC) procedure and the rapid insulin sensitivity test (RIST) to characterize insulin sensitivity in anaesthetized rats. The changes in insulin sensitivity were then supplemented with the direct measurement of insulin-stimulated glucose uptake using tissue accumulation of radioactive 2-deoxyglucose in skeletal muscle samples obtained from animals undergone either procedure. Studies of the recently described endogenous insulin sensitizer mechanism termed hepatic insulin sensitizing (HISS) mechanism, by the two methods yielded data for evaluation. The HISS mechanism is defined as an increase in tissue insulin sensitivity in response to post-prandial hepatic release of an undefined substance through a nitrergic pathway. For the HEGC method, insulin was infused to attain a stable plasma insulin immunoreactivity of 100 microU/ml determined by radioimmunoassay, whereas with the RIST method the HISS mechanism was activated by a 50 mg/kg i.v. insulin bolus. Euglycaemia was kept constant by means of glucose infusion. With the HEGC and the RIST methods, insulin sensitivity was defined as the average rate of glucose infusion and the amount of glucose/kg body weight/40 min (RIST index) infused to maintain euglycaemia and preinvestigation blood glucose level, respectively. During HEGC 16+/-4.2 mg/kg/min glucose was able to maintain euglycaemia, which decreased to 8+/-2.9 (p<0.05) after administration of 10 mg/kg NG-nitro-L-arginine methyl ester (L-NAME) (i.p.), a NO synthase inhibitor. Conversely, the RIST index decreased by 55+/-6.9% (p<0.05) after L-NAME. Similarly, 2-deoxyglucose uptake by the gastrocnemius muscle was decreased by 49.9+/-5.8 (p<0.05) and 52.3+/-7.4% (p<0.05) with the HEGC and the RIST methods, respectively. The results show that both the HEGC and the RIST methods supplemented with tissue radioactive 2-deoxyglucose uptake determinations are appropriate methods to characterize the alteration of insulin sensitivity in context of the HISS mechanism.

BGP-15 - a novel poly(ADP-ribose) polymerase inhibitor - protects against nephrotoxicity of cisplatin without compromising its antitumor activity.

Nephrotoxicity is one of the major dose limiting side effects of cisplatin chemotherapy. The antitumor and toxic effects are mediated in part by different mechanisms, thus, permitting a selective inhibition of certain side effects. The influence of O-(3-piperidino-2-hydroxy-1-propyl)nicotinic amidoxime (BGP-15) - a poly(ADP-ribose) polymerase (PARP) inhibitor - on the nephrotoxicity and antitumor efficacy of cisplatin has been evaluated in experimental models. BGP-15 either blocked or significantly reduced (60-90% in 100-200 mg/kg oral dose) cisplatin induced increase in serum urea and creatinine level in mice and rats and prevented the structural degeneration of the kidney, as well. The nephroprotective effect of BGP-15 treatment was revealed also in living mice by MRI analysis manifesting in the lack of oedema which otherwise developed as a result of cisplatin treatment. The protective effect was accompanied by inhibition of cisplatin-induced poly-ADP-ribosylation and by the restoration of the disturbed energy metabolism. The preservation of ATP level in the kidney was demonstrated in vivo by localized NMR spectroscopy. BGP-15 decreased cisplatin-induced ROS production in rat kidney mitochondria and improved the antioxidant status of the kidney in mice with cisplatin-induced nephropathy. In rat kidney, cisplatin caused a decrease in the level of Bcl-x, a mitochondrial protective protein, and this was normalized by BGP-15 treatment. On the other hand, BGP-15 did not inhibit the antitumor efficacy of cisplatin in cell culture and in transplantable solid tumors of mice. Treatment with BGP-15 increased the mean survival time of cisplatin-treated P-388 leukemia bearing mice from 13 to 19 days. PARP inhibitors have been demonstrated to diminish the consequences of free radical-induced damage, and this is related to the chemoprotective effect of BGP-15, a novel PARP inhibitor. Based on these results, we propose that BGP-15 represents a novel, non-thiol chemoprotective agent.