Stimulatory Effect of Puerarin on α_1A-Adrenoceptor to Increase Glucose Uptake into Cultured C₂C₁₂ Cells of Mice

Abstract

Effects of puerarin, an active principle contained in the roots of Pueraria lobata (Leguminosae), on the regulation of glucose metabolism in an insulin deficient state were investigated in cultured myoblast C₂C₁₂ cells using glucose uptake as indicator. Puerarin enhanced the uptake of radioactive glucose into C₂C₁₂ cells in a concentration-dependent manner, which was abolished by prazosin pretreatment. Activation of α₁-adrenoceptors by puerarin was further indicated by the displacement of [³ H]prazosin binding in C₂C₁₂ cells. The stimulatory action of puerarin on glucose uptake was also reduced in C₂C₁₂ cells pre-incubated with the antagonists, both WB 4101 and RS 17 056, at concentrations sufficient to block α_1A-adrenoceptor (α_1A-AR). An activation of α_1A-AR seems responsible for the action of puerarin in C₂C₁₂ cells. Pharmacological inhibition of phospholipase C (PLC) by U73312 resulted a concentration-dependent decrease of puerarin-stimulated glucose uptake in C₂C₁₂ cells. This inhibition of glucose uptake by U73122 was specific because the inactive congener, U73343, failed to block puerarin-stimulated glucose uptake. Moreover, both chelerythrine and GF 109203X diminished the action of puerarin at concentration sufficient to inhibit protein kinase C (PKC). The obtained data suggest that an activation of α_1A-AR by puerarin in C₂C₁₂ cells may increase the glucose uptake via the PLC-PKC pathway.

Key words

Puerarin - Pueraria lobata - Leguminosae - C₂C₁₂ cells - α_1A-adrenoceptor - phospholipase C - protein kinase C

References

• 1 Hieble J P, Bylund D B, Clarke D E, Eikenburg D C, Langer S Z, Lefkowitz R J, Minneman K P, Ruffolo RR J r. International Union of Pharmacology. X. Recommendation for nomenclature of alpha 1-adrenoceptors: consensus update.  Pharmacol Rev. 1995;  47 267-70
• 2 Faintrenie G, Géloén A. J. Alpha-1 adrenergic stimulation of glucose uptake in rat white adipocytes.  Pharmacol Exp Ther. 1998;  286 607-10
• 3 DeFronzo R A. The triumvirate: β-cell, muscle, liver. A collusion responsible for NIDDM.  Diabetes. 1988;  37 667-87
• 4 Cheng J T, Liu I M. Stimulatory effect of caffeic acid on α_1A-adrenoceptors to increase glucose uptake into cultured C₂C₁₂ cells.  Naunyn-Schmiedebergs Arch Pharmacol. 2000;  362 122-7
• 5 Liu I M, Tsai C C, Lai T Y, Cheng J T. Stimulatory effect of isoferulic acid on alpha1A-adrenoceptor to increase glucose uptake into cultured myoblast C2C12 cell of mice.  Auton Neurosci. 2001;  88 175-80
• 6 Kaufman P B, Duke J A, Brielmann H, Boik J, Hoyt J E. A comparative survey ofleguminous plants as sources of the isoflavones, genistein and daidzein: implications for human nutrition and health.  J Altern Complement Med. 1997;  3 7-12
• 7 Kim D H, Yu K U, Bae E A, Han M J. Metabolism of puerarin and daidzin by human intestinal bacteria and their relation to in vitro cytotoxicity.  Biol Pharm Bull. 1998;  21 628-30
• 8 Shen Z F, Xie M Z. Hypoglycemic effect of the combined use of puerarin and aspirin in mice.  Acta Pharm Sin. 1985;  20 863-6
• 9 Fan L L, O’Keefe D D, Powell WJ J r. Effect of puerarin on regional myocardial blood flow and cardiac hemodynamics in dogs with acute myocardial ischemia.  Acta Pharm Sin. 1984;  19 801-7
• 10 Szymanska G, Kim H W, Cuppoletti J, Kranias E G. Regulation of the skeletal sarcoplasmic reticulum Ca²⁺-ATPase by phospholamban and negatively charged phospholipids in reconstituted phospholipid vesicles.  Mol Cell Biochem. 1992;  114 65-71
• 11 Crist G H, Xu B, Lanoue F, Lang C H. Tissue-specific effects of in vivo adenosine receptor blocked on glucose uptake in Zucker rats.  FASEB J. 1998;  12 1301-8
• 12 Liu I M, Huang L W, Cheng J T. Gene expression of α_1A-adrenoceptor but not α_1B-adrenoceptor in cultured C₂C₁₂ cells.  Neurosci Lett. 2000;  294 93-6
• 13 Marshall I, Burt R P, Green G M, Hussain M B, Chapple C R. Different subtypes of α 1A-adrenoceptor mediating contraction of rat epididymal vas deferens, rat hepatic portal vein and human prostate distinguished by the antagonist RS 17 053.  Br J Pharmacol. 1996;  119 407-15
• 14 Piascik M T, Guarino R D, Smith M S, Soltis E E, Saussy DL J r, Perez D M. The specific contribution of the novel α-1D adrenoceptor to the contraction of vascular smooth muscle.  J Pharmacol Exp Ther. 1995;  275 1583-9
• 15 Ishizuka T, Cooper D R, Hernandez H, Buckley D, Standaert M, Farese R V. Effects of insulin on diacylglycerol-protein kinase C signaling in rat diaphragm and soleus muscles and relationship to glucose transport.  Diabetes. 1990;  39 181-90
• 16 Ploug T H, Galbo H, Richter E A. Increased muscle glucose uptake during contractions: no need for insulin.  Am J Physiol. 1984;  247 E726-31
• 17 Smallridge R C, Kiang J G, Gist I D, Fein H G, Gallowat R J. U-73 122, an aminosteroid phospholipase C antagonist, noncompetitively inhibits thyrotropin-releasing hormone effects in GH₃ rat pituitary cell.  Endocrinology. 1992;  131 1883-8
• 18 Toullec D, Pianetti P, Coste H, Bellevergue P, Grand-Perret T, Ajakane M. et al . The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C.  J Biol Chem. 1991;  266 15 771-81
• 19 Yang G, Zhang L, Fan L. Anti-angina effect of puerarin and its effect on plasma thromboxane A2 and prostacyclin.  Chinese J Integ Trad West Med. 1990;  10 82-4
• 20 Song X P, Chen P P, Chai X S. Effects of puerarin on blood pressure and plasma renin activity in spontaneously hypertensive rats.  Acta Pharm Sin. 1988;  9 55-8