Differential Metabolic Effects of Novel Cilostamide Analogs, Methyl Carbostiryl Derivatives, on Mouse and Hyperglycemic Rat

Document Type : Original Article


Pharmacological Research Centre of Medicinal Plants, Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


PDE3 has a functional role in insulin secretion and action. We investigated the metabolic effects of new synthetic PDE3 inhibitors (mc1, mc2, mc5 and mc6), on mice and hyperglycemic rat.
Materials and Methods
The test compound or solvent was injected subcutaneously to mice, for 7 days. On day 8, blood and liver samples were obtained. In hyperglycemic rat, 0.5 g/kg glucose with or without test compounds was injected, and followed with infusion of 1.5 g/kg/hr glucose. Blood samples were collected in mentioned intervals and liver was dissected.
In hyperglycemic rat, all test compounds decreased blood glucose and the effect of milrinone was potentiated by glybenclamide. Milrinone or IBMX did not change plasma insulin levels, but it was augmented by combination of milrinone and glybenclamide. In both species, liver glycogen storage was decreased by IBMX, mc5, mc6 or MCPIP, increased by mc2 (liver glycogen, rat, control=56±2, mc2=70±3 P< 0.01, mice, control=33±0.7, mc2=42±2.3 P< 0.01) and was not changed in the presence of mc1. Milrinone did not change the glycogen storage in rats though increased it in mice (control= 33±0.7, milrinone= 40±1 P< 0.05).
Increasing plasma insulin levels by combination of milrinone and glybenclamide confirmed that in hyperglycemic rat, the hypoglycemic effect was correlated with increasing insulin secretion. Variations of plasma insulin were obscured by the pulsative characteristic of pancreatic insulin release. Decreasing glycogen storage reflected inhibition of liver PDE activity. The reasons for ineffectiveness of mc1, anabolic effect of mc2, and differential effects of milrinone were not clear.


1. Degerman EE, Manganiello V, Ahren B. Milrinone efficiently potentiates insulin secretion induced by orally but not intravenously administered glucose in C57BL6J mice. Eur J Pharmacol 2004; 98:319-323.
2. Shafiee-Nick R, Pyne NJ, Furman BL. Effects of type-selective phosphodiesterase inhibitors on glucose-induced insulin secretion and islet phosphodiesterase activity. Br J Pharmacol 1995; 115:1486-1492.
3. Harndahl L, Jing X-J, Iverson R, Degerman E, Ahrén B, Manganiello VC, et al.Important role of phosphodiesrerase3B for the stimulatory action of cAMP on pancreatic β-cell exocytosis and release of insulin. J Biol Chem 2002; 277:37446-37455.
4. Zumuda -Trzebiatowska E, Oknianska A, Manganiello V. Role of DE3B in insulin-induced glucose uptake, GLU-T4 translocation and lipogenesis in primary rat adipocytes. Cell Signal 2006; 18:382-390.
5. Maurice DH, Palmer D, Tilley D, Dunkerley HA, Netherton SJ, Raymond DR, et al. Cyclic nucleotide phosphodiesterase activity, expression and targeting in cells of the cardiovascular system. Mol Pharmacol 2003; 64:533-536.
6.  Pyne NJ, Furman BL. Cyclic nucleotide phosphodiesterases in pancreatic islets. Diabetologia 2003; 46:1179-1189.
7. Cheung P, Yang G, Boden G.Milrinone a selective phosphodiesterase3 inhibitor stimulates lipolysis, endogenous glucose production and insulin secretion. Metabolism 2003; 52:1496-1500.
8. Parizadeh SMR, Shafiee Nik R, Zahraei M. Comparison between the insulinotropic effects of milrinone and amrinone, selective PDE3 inhibitors in in-vitro and in-vivo conditions. Iran J Basic Med Sci 2001; 9:7-15. 
9. Mansouri MT, Shafiee-Nick R, Parsaee H, Seyedi SM, Saberi MR, Sadeghian H. Inotropic and chronotropic effects of 6-hydroxy-4-methylquinolinone-2 (1H)-one derivatives in isolated rat atria. Iran Biomed J 2008; 12:77-84.
10. Sadeghian H, Seyedi M, Saberi MR, Nick RS, Hosseini A, Bakavoli M, et al. Design, synthesis and pharmacological evaluation of 6-hydroxy-4-methylquinolin-2 (1H)-one derivatives as inotropic agents. J Enzyme Inhib Med Chem 2008; 24:918-929.
11. Pour Ali Behzad N, Shafiee Nick R, Parizadeh SMR. Anti-hyperglycemic effects of cyclic nucleotides phosphodiesterases (PDEs) in rat. Med J Tabriz Univ Med Sci 2007; 29:11-15.
12. Waddleton D, Wub W, Feng Y, Thompson C, Wu M, Zhou YP, et al. Phosphodiesterase 3 and 4 comprise the major cAMP metabolizing enzymes responsible for insulin secretion in INS-1 (832/13) cells and rat islets. Biochem Pharmacol 2008; 76:884-893.
13. Ahmad M, Abdel-Wahab YHA, Tate R,  Flatt PR, Pyne NJ, Furman BL. Effect of type-selective inhibitors on cyclic nucleotide phosphodiesterase activity and insulin secretion in the clonal insulin secreting cell line BRIN-BD11. Br J Pharmacol 2000; 129:1228-1234.
14. EL-Metwally M, Shafiee-Nick R, Pyne NJ, Furman BL. Effects of Org 9935, a selective type III phosphodiesterase inhibitor and Org 30029 a mixed type III/IV phosphodiesterase inhibitor on glucose-induced insulin secretion in vivo and in vitro. Eur J Pharmacol 1997; 324:227- 232.
15. Yang G, Li L. In vivo effects of phosphodiesterase III inhibitors on glucose metabolism and insulin sensitivity. J Chin Med Assoc 2003; 66:210– 226.
16. Parker JC, VanVolkenburg MA, Nardone NA, Hargrove DM, Andrews KM. Modulation of insulin secretion and glycemia by selective inhibition of cyclic AMP phosphodiesterase III. Biochem Biophys Res Commun 1997; 236:665– 669.
17. Furman BL, Pyne NJ. Islet phosphodiesterase isoenzymes and insulin secretion. Diabetic Med 1990; 7:19A.
18. Zmuda-Trzebiatowskaa E, Oknianskaa A, Manganiellob V, Degerman E.  Role of PDE3B in insulin-induced glucose uptake, GLUT-4 translocation`` and lipogenesis in primary rat adipocytes. Cell Signal  2006; 18:382 – 390.
19. Beebe SJ, Redmon JB, Blackmore PF, Corbin JD. Discriminative insulin antagonism of stimulatory effects of various cAMP analogs on adipocyte lipolysis and hepatocytes glycogenolysis. J Biol Chem 1985; 260:15781– 15788.
20. Arner P. Insulin resistance in type 2 diabetes: role of fatty acids. Diabetes Metab Res Rev 2002; 18: S5–S9.
21. Boden G. Effects of free fatty acids (FFA) on glucose metabolism: significance for insulin resistance and type 2 diabetes. Exp Clin Endocrine Diabetes 2003; 111:121–124.
22. Eriksson H, Ridderstrale M, Degerman E, Ekholm D, Smith CJ, Manganiello VC, et al. Evidence for the key role of the adipocyte cGMP-inhibited cAMP phosphodiesterase in the antilipolytic action of insulin. Biochem Biophys Acta 1995; 1266:101–107.
23. Hagstrfm-Toft E, Bolinder J, Eriksson S, Arner P. Role of phosphodiesterase III in the antilipolytic effect of insulin in vivo. Diabetes 1995; 44:1170– 1175.
24. Zhao AZ, Karin A, Beavo JA. Leptin inhibits insulin secretion by activation of PDE3B. J Clin Invest 1998; 102: 869-873.
25. Yamada S, Komatsu M, Sato Y, Yamauchi K, Kojima I. Time dependent stimulation of insulin exocytosis by 3, 5 cyclic adenosine monophosphate in the rat islet β-cells. Endocrinology 2002; 143: 4203-4209.
26. Doyle M, Egan J.Pharmacological agents that directly modulate insulin secretion. Pharmacol Rev 2006; 55:105-131.
27. Porksen N, Hollingdal M, Juhl C, Butler P, Veldhuis JD, Schmitz O. Pulsatile insulin secretion: detection,  egulation, and role in diabetes. Diabetes 2002; 51:245-254.
28. Parker JC, Volkenburg MA, Gao Feng. Synergistic effect on insulin secretion from INS-1 cells of a sulfonylurea and a phosphodiesterase 3 inhibitor. Life Sci 2004; 75: 1479–1490.
29. Bollen M, Keppens S, Stalmans W. Specific features of glycogen metabolism in the liver. Biochem J 1998; 336:19-31.
30. Fong M, Yoshitake M, Kambayashi J, Liu Y. Cilostazol increases tissue blood flow in contracting rabbit gastrocnemius muscle. Circ J 2010; 74:181-187.
31. Baron AD, Steinberg H, Brechtel G, Johnson A. Skeletal muscle blood flow independently modulates insulin-mediated glucose uptake. Am J Physiol 1994; 266: E248–E253.
32. Baron AD, Steinberg HO, Chaker H, Irsula O, Brechtel G, Keech C. Insulin-mediated skeletal muscle vasodilatation contributes to both insulin sensitivity and responsiveness in lean humans. J Clin Invest 1995; 96:786–792.
33. Weishaar RE, Kobylarz-Singer DC, Steffen RP, , Kaplan HR. Subclasses of cyclic AMP-specific phosphodiesterase in left ventricular muscle and their involvement in regulating myocardial contractility. Circ  Res 1987; 61:539-547.
34. Laurberg P. Forskolin stimulation of thyroid secretion of T4 and T3. FEBS 1984; 170:273-276.
35. Schery MP, Brown BL, Ekins RP. Studies on the role of calcium and cyclic nucleotide in the control of TSH secretion. Mol Cell Endocrine 1978; 11:249-264.
36.Klieverik LP, Janssena SF, Riel A, Foppen E, Bisschop PH, Serlie MJ, et al. Thyroid hormone modulates glucose production via a sympathetic pathway from the hypothalamic paraventricular nucleus to the liver. Proc Natl Acad Sci U S A 2009; 106:5966–5971.