Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Sep;92(3):1161–1167. doi: 10.1172/JCI116685

Insulin attenuates agonist-mediated calcium mobilization in cultured rat vascular smooth muscle cells.

F Saito 1, M T Hori 1, M Fittingoff 1, T Hino 1, M L Tuck 1
PMCID: PMC288253  PMID: 8397220

Abstract

Insulin has been shown to attenuate pressor-induced vascular contraction, but the mechanism for this vasodilatory action is unknown. This study examines the effect of insulin on angiotensin II (ANG II)-induced increments in cytosolic calcium in cultured rat vascular smooth muscle cells (VSMC). 20-min incubations with insulin (10 microU/ml to 100 mU/ml) did not alter basal intracellular calcium concentration ([Ca2+]i), but inhibited the response to 100 nM ANG II in a dose-dependent manner (ANG II alone, 721 +/- 54 vs. ANG II + 100 mU/ml insulin, 315 +/- 35 nM, P < 0.01). A similar effect of insulin on ANG II action was observed in calcium poor buffer. Moreover, insulin did not effect calcium influx. ANG II receptor density and affinity were not affected by 24-h incubation with insulin. To further clarify the mechanisms of these observations, we measured ANG II-induced production of inositol 1,4,5-triphosphate (IP3), and IP3-releasable 45Ca. Insulin treatment did not alter ANG II-stimulated IP3 production. However, IP3-stimulated release of 45Ca in digitonin permeabilized cells was significantly reduced after 5-min incubations with 100 mU/ml insulin. Thapsigargin induced release of calcium stores was also blocked by insulin. Thus, insulin attenuates ANG II-stimulated [Ca2+]i primarily by altering IP3-releasable calcium stores. Insulin effects on ANG II-induced [Ca2+]i were mimicked by preincubation of VSMC with either sodium nitroprusside or 8-bromo-cGMP. As elevations in cGMP in vascular tissue lower [Ca2+]i, it is possible that insulin affects IP3 release of calcium by a cGMP-dependent mechanism that would contribute to its vasodilatory effects.

Full text

PDF
1161

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alexander W. D., Oake R. J. The effect of insulin on vascular reactivity to norepinephrine. Diabetes. 1977 Jul;26(7):611–614. doi: 10.2337/diab.26.7.611. [DOI] [PubMed] [Google Scholar]
  2. Bar R. S., Boes M., Dake B. L., Booth B. A., Henley S. A., Sandra A. Insulin, insulin-like growth factors, and vascular endothelium. Am J Med. 1988 Nov 28;85(5A):59–70. doi: 10.1016/0002-9343(88)90398-1. [DOI] [PubMed] [Google Scholar]
  3. Bernhardt J., Tschudi M. R., Dohi Y., Gut I., Urwyler B., Bühler F. R., Lüscher T. F. Release of nitric oxide from human vascular smooth muscle cells. Biochem Biophys Res Commun. 1991 Oct 31;180(2):907–912. doi: 10.1016/s0006-291x(05)81151-9. [DOI] [PubMed] [Google Scholar]
  4. Blaustein M. P. Sodium/calcium exchange and the control of contractility in cardiac muscle and vascular smooth muscle. J Cardiovasc Pharmacol. 1988;12 (Suppl 5):S56–S68. [PubMed] [Google Scholar]
  5. Brock T. A., Lewis L. J., Smith J. B. Angiotensin increases Na+ entry and Na+/K+ pump activity in cultures of smooth muscle from rat aorta. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1438–1442. doi: 10.1073/pnas.79.5.1438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Calviello G., Chiesi M. Rapid kinetic analysis of the calcium-release channels of skeletal muscle sarcoplasmic reticulum: the effect of inhibitors. Biochemistry. 1989 Feb 7;28(3):1301–1306. doi: 10.1021/bi00429a053. [DOI] [PubMed] [Google Scholar]
  7. Casteels R., Raeymaekers L., Droogmans G., Wuytack F. Na+-K+ ATPase, Na-Ca exchange, and excitation-contraction coupling in smooth muscle. J Cardiovasc Pharmacol. 1985;7 (Suppl 3):S103–S110. [PubMed] [Google Scholar]
  8. Clausen T., Martin B. R. The effect of insulin on the washout of [45Ca]calcium from adipocytes and soleus muscle of the rat. Biochem J. 1977 Apr 15;164(1):251–255. doi: 10.1042/bj1640251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Degerman E., Smith C. J., Tornqvist H., Vasta V., Belfrage P., Manganiello V. C. Evidence that insulin and isoprenaline activate the cGMP-inhibited low-Km cAMP phosphodiesterase in rat fat cells by phosphorylation. Proc Natl Acad Sci U S A. 1990 Jan;87(2):533–537. doi: 10.1073/pnas.87.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Desnuelle C., Askanas V., Engel W. K. Insulin enhances development of functional voltage-dependent Ca2+ channels in aneurally cultured human muscle. J Neurochem. 1987 Oct;49(4):1133–1138. doi: 10.1111/j.1471-4159.1987.tb10003.x. [DOI] [PubMed] [Google Scholar]
  11. Downing S. E., Lee J. C., Matisoff D. N. Coronary blood flow in the diabetic lamb with metabolic acidosis. Am J Physiol. 1980 Feb;238(2):H263–H268. doi: 10.1152/ajpheart.1980.238.2.H263. [DOI] [PubMed] [Google Scholar]
  12. Draznin B., Kao M., Sussman K. E. Insulin and glyburide increase cytosolic free-Ca2+ concentration in isolated rat adipocytes. Diabetes. 1987 Feb;36(2):174–178. doi: 10.2337/diab.36.2.174. [DOI] [PubMed] [Google Scholar]
  13. Fittingoff M., Krall J. F. Changes in inositol polyphosphate-sensitive calcium exchange in aortic smooth muscle cells in vitro. J Cell Physiol. 1988 Feb;134(2):297–301. doi: 10.1002/jcp.1041340218. [DOI] [PubMed] [Google Scholar]
  14. Flatman J. A., Clausen T. Combined effects of adrenaline and insulin on active electrogenic Na+-K+ transport in rat soleus muscle. Nature. 1979 Oct 18;281(5732):580–581. doi: 10.1038/281580a0. [DOI] [PubMed] [Google Scholar]
  15. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  16. Gunther S., Alexander R. W., Atkinson W. J., Gimbrone M. A., Jr Functional angiotensin II receptors in cultured vascular smooth muscle cells. J Cell Biol. 1982 Feb;92(2):289–298. doi: 10.1083/jcb.92.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gupta M. P., Makino N., Khatter K., Dhalla N. S. Stimulation of Na+-Ca2+ exchange in heart sarcolemma by insulin. Life Sci. 1986 Sep 22;39(12):1077–1083. doi: 10.1016/0024-3205(86)90199-2. [DOI] [PubMed] [Google Scholar]
  18. Head R. J., Longhurst P. A., Panek R. L., Stitzel R. E. A contrasting effect of the diabetic state upon the contractile responses of aortic preparations from the rat and rabbit. Br J Pharmacol. 1987 Jun;91(2):275–286. doi: 10.1111/j.1476-5381.1987.tb10282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heyworth C. M., Houslay M. D. Insulin exerts actions through a distinct species of guanine nucleotide regulatory protein: inhibition of adenylate cyclase. Biochem J. 1983 Aug 15;214(2):547–552. doi: 10.1042/bj2140547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kurtz T. W., Morris R. C., Pershadsingh H. A. The Zucker fatty rat as a genetic model of obesity and hypertension. Hypertension. 1989 Jun;13(6 Pt 2):896–901. doi: 10.1161/01.hyp.13.6.896. [DOI] [PubMed] [Google Scholar]
  21. Laakso M., Edelman S. V., Brechtel G., Baron A. D. Decreased effect of insulin to stimulate skeletal muscle blood flow in obese man. A novel mechanism for insulin resistance. J Clin Invest. 1990 Jun;85(6):1844–1852. doi: 10.1172/JCI114644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Levy J., Gavin J. R., 3rd, Hammerman M. R., Avioli L. V. Ca2+-Mg2+-ATPase activity in kidney basolateral membrane in non-insulin-dependent diabetic rats. Effect of insulin. Diabetes. 1986 Aug;35(8):899–905. doi: 10.2337/diab.35.8.899. [DOI] [PubMed] [Google Scholar]
  23. Levy J., Gavin J. R., 3rd, Morimoto S., Hammerman M. R., Avioli L. V. Hormonal regulation of (Ca2+ + Mg2+)ATPase activity in canine renal basolateral membrane. Endocrinology. 1986 Dec;119(6):2405–2411. doi: 10.1210/endo-119-6-2405. [DOI] [PubMed] [Google Scholar]
  24. Liang C., Doherty J. U., Faillace R., Maekawa K., Arnold S., Gavras H., Hood W. B., Jr Insulin infusion in conscious dogs. Effects on systemic and coronary hemodynamics, regional blood flows, and plasma catecholamines. J Clin Invest. 1982 Jun;69(6):1321–1336. doi: 10.1172/JCI110572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lytton J., Westlin M., Hanley M. R. Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. J Biol Chem. 1991 Sep 15;266(26):17067–17071. [PubMed] [Google Scholar]
  26. McDaniel N. L., Chen X. L., Singer H. A., Murphy R. A., Rembold C. M. Nitrovasodilators relax arterial smooth muscle by decreasing [Ca2+]i and uncoupling stress from myosin phosphorylation. Am J Physiol. 1992 Aug;263(2 Pt 1):C461–C467. doi: 10.1152/ajpcell.1992.263.2.C461. [DOI] [PubMed] [Google Scholar]
  27. Modan M., Halkin H., Almog S., Lusky A., Eshkol A., Shefi M., Shitrit A., Fuchs Z. Hyperinsulinemia. A link between hypertension obesity and glucose intolerance. J Clin Invest. 1985 Mar;75(3):809–817. doi: 10.1172/JCI111776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  29. Oba T., Iwama H., Aoki T. Ruthenium red and magnesium ion partially inhibit silver ion-induced release of calcium from sarcoplasmic reticulum of frog skeletal muscles. Jpn J Physiol. 1989;39(2):241–254. doi: 10.2170/jjphysiol.39.241. [DOI] [PubMed] [Google Scholar]
  30. Page M. M., Watkins P. J. Provocation of postural hypotension by insulin in diabetic autonomic neuropathy. Diabetes. 1976 Feb;25(2):90–95. doi: 10.2337/diab.25.2.90. [DOI] [PubMed] [Google Scholar]
  31. Pershadsingh H. A., McDonald J. M. Direct addition of insulin inhibits a high affinity Ca2+-ATPase in isolated adipocyte plasma membranes. Nature. 1979 Oct 11;281(5731):495–497. doi: 10.1038/281495a0. [DOI] [PubMed] [Google Scholar]
  32. Pfaffman M. A., Ball C. R., Darby A., Hilman R. Insulin reversal of diabetes-induced inhibition of vascular contractility in the rat. Am J Physiol. 1982 Apr;242(4):H490–H495. doi: 10.1152/ajpheart.1982.242.4.H490. [DOI] [PubMed] [Google Scholar]
  33. Pfeifle B., Ditschuneit H. Receptors for insulin and insulin-like growth factor in cultured arterial smooth muscle cells depend on their growth state. J Endocrinol. 1983 Feb;96(2):251–257. doi: 10.1677/joe.0.0960251. [DOI] [PubMed] [Google Scholar]
  34. Pfeifle B., Ditschuneit H. Two separate receptors for insulin and insulinlike growth factors on arterial smooth muscle cells. Exp Clin Endocrinol. 1983 May;81(3):280–286. doi: 10.1055/s-0029-1210237. [DOI] [PubMed] [Google Scholar]
  35. Pyne N. J., Houslay M. D. An insulin mediator preparation serves to stimulate the cyclic GMP activated cyclic AMP phosphodiesterase rather than other purified insulin-activated cyclic AMP phosphodiesterases. Biochem Biophys Res Commun. 1988 Oct 14;156(1):290–296. doi: 10.1016/s0006-291x(88)80838-6. [DOI] [PubMed] [Google Scholar]
  36. Reaven G. M., Ho H., Hoffmann B. B. Somatostatin inhibition of fructose-induced hypertension. Hypertension. 1989 Aug;14(2):117–120. doi: 10.1161/01.hyp.14.2.117. [DOI] [PubMed] [Google Scholar]
  37. Resink T. J., Tkachuk V. A., Erne P., Bühler F. R. Platelet membrane calmodulin-stimulated calcium-adenosine triphosphatase. Altered activity in essential hypertension. Hypertension. 1986 Feb;8(2):159–166. doi: 10.1161/01.hyp.8.2.159. [DOI] [PubMed] [Google Scholar]
  38. Rocchini A. P., Key J., Bondie D., Chico R., Moorehead C., Katch V., Martin M. The effect of weight loss on the sensitivity of blood pressure to sodium in obese adolescents. N Engl J Med. 1989 Aug 31;321(9):580–585. doi: 10.1056/NEJM198908313210905. [DOI] [PubMed] [Google Scholar]
  39. Rose H. G., Yalow R. S., Schweitzer P., Schwartz E. Insulin as a potential factor influencing blood pressure in amputees. Hypertension. 1986 Sep;8(9):793–800. doi: 10.1161/01.hyp.8.9.793. [DOI] [PubMed] [Google Scholar]
  40. Rowe J. W., Young J. B., Minaker K. L., Stevens A. L., Pallotta J., Landsberg L. Effect of insulin and glucose infusions on sympathetic nervous system activity in normal man. Diabetes. 1981 Mar;30(3):219–225. doi: 10.2337/diab.30.3.219. [DOI] [PubMed] [Google Scholar]
  41. Sato K., Ozaki H., Karaki H. Changes in cytosolic calcium level in vascular smooth muscle strip measured simultaneously with contraction using fluorescent calcium indicator fura 2. J Pharmacol Exp Ther. 1988 Jul;246(1):294–300. [PubMed] [Google Scholar]
  42. Simpson A. W., Stampfl A., Ashley C. C. Evidence for receptor-mediated bivalent-cation entry in A10 vascular smooth-muscle cells. Biochem J. 1990 Apr 1;267(1):277–280. doi: 10.1042/bj2670277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Somlyo A. V., Bond M., Somlyo A. P., Scarpa A. Inositol trisphosphate-induced calcium release and contraction in vascular smooth muscle. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5231–5235. doi: 10.1073/pnas.82.15.5231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sowers J. R., Whitfield L. A., Catania R. A., Stern N., Tuck M. L., Dornfeld L., Maxwell M. Role of the sympathetic nervous system in blood pressure maintenance in obesity. J Clin Endocrinol Metab. 1982 Jun;54(6):1181–1186. doi: 10.1210/jcem-54-6-1181. [DOI] [PubMed] [Google Scholar]
  45. Standley P. R., Zhang F., Ram J. L., Zemel M. B., Sowers J. R. Insulin attenuates vasopressin-induced calcium transients and a voltage-dependent calcium response in rat vascular smooth muscle cells. J Clin Invest. 1991 Oct;88(4):1230–1236. doi: 10.1172/JCI115426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Swislocki A. L., Hoffman B. B., Reaven G. M. Insulin resistance, glucose intolerance and hyperinsulinemia in patients with hypertension. Am J Hypertens. 1989 Jun;2(6 Pt 1):419–423. doi: 10.1093/ajh/2.6.419. [DOI] [PubMed] [Google Scholar]
  47. Takata S., Yamamoto M., Yagi S., Noto Y., Ikeda T., Hattori N. Peripheral circulatory effects of insulin in diabetes. Angiology. 1985 Feb;36(2):110–115. doi: 10.1177/000331978503600207. [DOI] [PubMed] [Google Scholar]
  48. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tuck M., Corry D., Trujillo A. Salt-sensitive blood pressure and exaggerated vascular reactivity in the hypertension of diabetes mellitus. Am J Med. 1990 Mar;88(3):210–216. doi: 10.1016/0002-9343(90)90144-3. [DOI] [PubMed] [Google Scholar]
  50. Vincenzi F. F., Morris C. D., Kinsel L. B., Kenny M., McCarron D. A. Decreased calcium pump adenosine triphosphatase in red blood cells of hypertensive subjects. Hypertension. 1986 Nov;8(11):1058–1066. doi: 10.1161/01.hyp.8.11.1058. [DOI] [PubMed] [Google Scholar]
  51. Wu F. S., Zierler K. Calcium currents in rat myoballs and their inhibition by insulin. Endocrinology. 1989 Nov;125(5):2563–2572. doi: 10.1210/endo-125-5-2563. [DOI] [PubMed] [Google Scholar]
  52. Yagi S., Takata S., Kiyokawa H., Yamamoto M., Noto Y., Ikeda T., Hattori N. Effects of insulin on vasoconstrictive responses to norepinephrine and angiotensin II in rabbit femoral artery and vein. Diabetes. 1988 Aug;37(8):1064–1067. doi: 10.2337/diab.37.8.1064. [DOI] [PubMed] [Google Scholar]
  53. Yanagisawa-Miwa A., Ito H., Sugimoto T. Effects of insulin on vasoconstriction induced by thromboxane A2 in porcine coronary artery. Circulation. 1990 May;81(5):1654–1659. doi: 10.1161/01.cir.81.5.1654. [DOI] [PubMed] [Google Scholar]
  54. Yokota M., Shimoda K., Kobayashi Y., Kajiwara N. [Insulin and hypertension--the mechanisms of high blood pressure during chronic insulin infusion]. Nihon Jinzo Gakkai Shi. 1989 Aug;31(8):875–881. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES