Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1992 Oct;90(4):1263–1268. doi: 10.1172/JCI115989

Prolonged exposure of human pancreatic islets to high glucose concentrations in vitro impairs the beta-cell function.

D L Eizirik 1, G S Korbutt 1, C Hellerström 1
PMCID: PMC443168  PMID: 1401063

Abstract

The aim of the present study was to clarify whether prolonged in vitro exposure of human pancreatic islets to high glucose concentrations impairs the function of these cells. For this purpose, islets isolated from adult cadaveric organ donors were cultured for seven days in RPMI 1640 medium supplemented with 10% fetal calf serum and containing either 5.6, 11, or 28 mM glucose. There was no glucose-induced decrease in islet DNA content or signs of morphological damage. However, islets cultured at 11 or 28 mM glucose showed a 45 or 60% decrease in insulin content, as compared to islets cultured at 5.6 mM glucose. Moreover, when such islets were submitted to a 60-min stimulation with a low (1.7 mM) followed by a high (16.7 mM) concentration of glucose, the islets cultured at 5.6 mM glucose showed a higher insulin response to glucose than those of the two other groups. Islets cultured at the two higher glucose concentrations showed increased rates of insulin release in the presence of low glucose, and a failure to enhance further the release in response to an elevated glucose level. Islets cultured at 28 mM glucose showed an absolute decrease in insulin release after stimulation with 16.7 mM glucose, as compared to islets cultured at 5.6 mM glucose. The rates of glucose oxidation, proinsulin biosynthesis, and total protein biosynthesis were similar in islets cultured at 5.6 or 11 mM glucose, but they were decreased in islets cultured at 28 mM glucose. These combined results suggest that lasting exposure to high glucose concentrations impairs the function of human pancreatic islets.

Full text

PDF
1263

Selected References

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

  1. Andersson A. Isolated mouse pancreatic islets in culture: effects of serum and different culture media on the insulin production of the islets. Diabetologia. 1978 Jun;14(6):397–404. doi: 10.1007/BF01228134. [DOI] [PubMed] [Google Scholar]
  2. Andersson A., Sandler S. Viability tests of cryopreserved endocrine pancreatic cells. Cryobiology. 1983 Apr;20(2):161–168. doi: 10.1016/0011-2240(83)90005-6. [DOI] [PubMed] [Google Scholar]
  3. Andersson A., Westman J., Hellerström C. Effects of glucose on the ultrastructure and insulin biosynthesis of isolated mouse pancreatic islets maintained in tissue culture. Diabetologia. 1974 Dec;10(6):743–753. doi: 10.1007/BF01219536. [DOI] [PubMed] [Google Scholar]
  4. Bolaffi J. L., Bruno L., Heldt A., Grodsky G. M. Characteristics of desensitization of insulin secretion in fully in vitro systems. Endocrinology. 1988 May;122(5):1801–1809. doi: 10.1210/endo-122-5-1801. [DOI] [PubMed] [Google Scholar]
  5. Bolaffi J. L., Heldt A., Lewis L. D., Grodsky G. M. The third phase of in vitro insulin secretion. Evidence for glucose insensitivity. Diabetes. 1986 Mar;35(3):370–373. doi: 10.2337/diab.35.3.370. [DOI] [PubMed] [Google Scholar]
  6. Bone A. J., Swenne I., Hellerström C. Effects of high glucose concentrations on the insulin biosynthesis of rat pancreatic islets maintained for extended periods in tissue culture. Diabete Metab. 1981 Dec;7(4):259–263. [PubMed] [Google Scholar]
  7. Buschard K. The functional state of the beta cells in the pathogenesis of insulin-dependent diabetes mellitus. Autoimmunity. 1991;10(1):65–69. doi: 10.3109/08916939108997149. [DOI] [PubMed] [Google Scholar]
  8. Davalli A. M., Ricordi C., Socci C., Braghi S., Bertuzzi F., Fattor B., Di Carlo V., Pontiroli A. E., Pozza G. Abnormal sensitivity to glucose of human islets cultured in a high glucose medium: partial reversibility after an additional culture in a normal glucose medium. J Clin Endocrinol Metab. 1991 Jan;72(1):202–208. doi: 10.1210/jcem-72-1-202. [DOI] [PubMed] [Google Scholar]
  9. Dohan F. C., Lukens F. D. Lesions of the Pancreatic Islets Produced in Cats by Administration of Glucose. Science. 1947 Feb 14;105(2720):183–183. doi: 10.1126/science.105.2720.183. [DOI] [PubMed] [Google Scholar]
  10. Eizirik D. L., Sandler S. Sustained exposure of toxically damaged mouse pancreatic islets to high glucose does not increase beta-cell dysfunction. J Endocrinol. 1989 Oct;123(1):47–51. doi: 10.1677/joe.0.1230047. [DOI] [PubMed] [Google Scholar]
  11. Eizirik D. L., Strandell E., Sandler S. Culture of mouse pancreatic islets in different glucose concentrations modifies B cell sensitivity to streptozotocin. Diabetologia. 1988 Mar;31(3):168–174. doi: 10.1007/BF00276851. [DOI] [PubMed] [Google Scholar]
  12. Eizirik D. L., Strandell E., Sandler S. Prolonged exposure of pancreatic islets isolated from "pre-diabetic" non-obese diabetic mice to a high glucose concentration does not impair beta-cell function. Diabetologia. 1991 Jan;34(1):6–11. doi: 10.1007/BF00404017. [DOI] [PubMed] [Google Scholar]
  13. Giroix M. H., Serradas P., Portha B. The desensitization of normal B-cells to glucose in vitro is transient and not related to high glucose levels. Endocrinology. 1989 Oct;125(4):1999–2007. doi: 10.1210/endo-125-4-1999. [DOI] [PubMed] [Google Scholar]
  14. Halban P. A., Wollheim C. B., Blondel B., Renold A. E. Long-term exposure of isolated pancreatic islets to mannoheptulose: evidence for insulin degradation in the beta cell. Biochem Pharmacol. 1980 Oct 1;29(19):2625–2633. doi: 10.1016/0006-2952(80)90077-5. [DOI] [PubMed] [Google Scholar]
  15. Heding L. G. Determination of total serum insulin (IRI) in insulin-treated diabetic patients. Diabetologia. 1972 Aug;8(4):260–266. doi: 10.1007/BF01225569. [DOI] [PubMed] [Google Scholar]
  16. Hinegardner R. T. An improved fluorometric assay for DNA. Anal Biochem. 1971 Jan;39(1):197–201. doi: 10.1016/0003-2697(71)90476-3. [DOI] [PubMed] [Google Scholar]
  17. Hoenig M., MacGregor L. C., Matschinsky F. M. In vitro exhaustion of pancreatic beta-cells. Am J Physiol. 1986 May;250(5 Pt 1):E502–E511. doi: 10.1152/ajpendo.1986.250.5.E502. [DOI] [PubMed] [Google Scholar]
  18. Imamura T., Koffler M., Helderman J. H., Prince D., Thirlby R., Inman L., Unger R. H. Severe diabetes induced in subtotally depancreatized dogs by sustained hyperglycemia. Diabetes. 1988 May;37(5):600–609. doi: 10.2337/diab.37.5.600. [DOI] [PubMed] [Google Scholar]
  19. Kaiser N., Corcos A. P., Sarel I., Cerasi E. Monolayer culture of adult rat pancreatic islets on extracellular matrix: modulation of B-cell function by chronic exposure to high glucose. Endocrinology. 1991 Oct;129(4):2067–2076. doi: 10.1210/endo-129-4-2067. [DOI] [PubMed] [Google Scholar]
  20. Komiya I., Baetens D., Kuwajima M., Orci L., Unger R. H. Compensatory capabilities of islets of BB/Wor rats exposed to sustained hyperglycemia. Metabolism. 1990 Jun;39(6):614–618. doi: 10.1016/0026-0495(90)90028-b. [DOI] [PubMed] [Google Scholar]
  21. Korsgren O., Jansson L., Sandler S., Andersson A. Hyperglycemia-induced B cell toxicity. The fate of pancreatic islets transplanted into diabetic mice is dependent on their genetic background. J Clin Invest. 1990 Dec;86(6):2161–2168. doi: 10.1172/JCI114955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kämpe O., Andersson A., Björk E., Hallberg A., Karlsson F. A. High-glucose stimulation of 64,000-Mr islet cell autoantigen expression. Diabetes. 1989 Oct;38(10):1326–1328. doi: 10.2337/diab.38.10.1326. [DOI] [PubMed] [Google Scholar]
  23. Madsbad S., Krarup T., Faber O. K., Binder C., Regeur L. The transient effect of strict glycaemic control on B cell function in newly diagnosed type 1 (insulin-dependent) diabetic patients. Diabetologia. 1982 Jan;22(1):16–20. doi: 10.1007/BF00253863. [DOI] [PubMed] [Google Scholar]
  24. Malaisse-Lagae F., Sener A., Malaisse W. J. Can desensitization of the B-cell to D-glucose be simulated in cultured pancreatic islets? Acta Diabetol Lat. 1987 Jan-Mar;24(1):17–25. doi: 10.1007/BF02732049. [DOI] [PubMed] [Google Scholar]
  25. Marynissen G., Leclercq-Meyer V., Sener A., Malaisse W. J. Perturbation of pancreatic islet function in glucose-infused rats. Metabolism. 1990 Jan;39(1):87–95. doi: 10.1016/0026-0495(90)90153-4. [DOI] [PubMed] [Google Scholar]
  26. Mirouze J., Selam J. L., Pham T. C., Mendoza E., Orsetti A. Sustained insulin-induced remissions of juvenile diabetes by means of an external artificial pancreas. Diabetologia. 1978 Apr;14(4):223–227. doi: 10.1007/BF01219420. [DOI] [PubMed] [Google Scholar]
  27. Portha B., Blondel O., Serradas P., McEvoy R., Giroix M. H., Kergoat M., Bailbe D. The rat models of non-insulin dependent diabetes induced by neonatal streptozotocin. Diabete Metab. 1989 Mar-Apr;15(2):61–75. [PubMed] [Google Scholar]
  28. Purrello F., Vetri M., Gatta C., Gullo D., Vigneri R. Effects of high glucose on insulin secretion by isolated rat islets and purified beta-cells and possible role of glycosylation. Diabetes. 1989 Nov;38(11):1417–1422. doi: 10.2337/diab.38.11.1417. [DOI] [PubMed] [Google Scholar]
  29. Rossetti L., Giaccari A., DeFronzo R. A. Glucose toxicity. Diabetes Care. 1990 Jun;13(6):610–630. doi: 10.2337/diacare.13.6.610. [DOI] [PubMed] [Google Scholar]
  30. Sako Y., Eizirik D., Grill V. Impact of uncoupling glucose stimulus from secretion on B-cell release and biosynthesis. Am J Physiol. 1992 Feb;262(2 Pt 1):E150–E154. doi: 10.1152/ajpendo.1992.262.2.E150. [DOI] [PubMed] [Google Scholar]
  31. Sako Y., Grill V. E. Coupling of beta-cell desensitization by hyperglycemia to excessive stimulation and circulating insulin in glucose-infused rats. Diabetes. 1990 Dec;39(12):1580–1583. doi: 10.2337/diab.39.12.1580. [DOI] [PubMed] [Google Scholar]
  32. Scharp D. W., Lacy P. E., Santiago J. V., McCullough C. S., Weide L. G., Falqui L., Marchetti P., Gingerich R. L., Jaffe A. S., Cryer P. E. Insulin independence after islet transplantation into type I diabetic patient. Diabetes. 1990 Apr;39(4):515–518. doi: 10.2337/diab.39.4.515. [DOI] [PubMed] [Google Scholar]
  33. Shah S. C., Malone J. I., Simpson N. E. A randomized trial of intensive insulin therapy in newly diagnosed insulin-dependent diabetes mellitus. N Engl J Med. 1989 Mar 2;320(9):550–554. doi: 10.1056/NEJM198903023200902. [DOI] [PubMed] [Google Scholar]
  34. Svensson C., Hellerström C. Long-term effects of a high glucose concentration in vitro on the oxidative metabolism and insulin production of isolated rat pancreatic islets. Metabolism. 1991 May;40(5):513–518. doi: 10.1016/0026-0495(91)90233-m. [DOI] [PubMed] [Google Scholar]
  35. Warnock G. L., Ellis D. K., Cattral M., Untch D., Kneteman N. M., Rajotte R. V. Viable purified islets of Langerhans from collagenase-perfused human pancreas. Diabetes. 1989 Jan;38 (Suppl 1):136–139. doi: 10.2337/diab.38.1.s136. [DOI] [PubMed] [Google Scholar]
  36. Warnock G. L., Kneteman N. M., Ryan E., Seelis R. E., Rabinovitch A., Rajotte R. V. Normoglycaemia after transplantation of freshly isolated and cryopreserved pancreatic islets in type 1 (insulin-dependent) diabetes mellitus. Diabetologia. 1991 Jan;34(1):55–58. doi: 10.1007/BF00404026. [DOI] [PubMed] [Google Scholar]
  37. Weir G. C., Leahy J. L., Bonner-Weir S. Experimental reduction of B-cell mass: implications for the pathogenesis of diabetes. Diabetes Metab Rev. 1986;2(1-2):125–161. doi: 10.1002/dmr.5610020108. [DOI] [PubMed] [Google Scholar]
  38. Welsh N., Hellerström C. In vitro restoration of insulin production in islets from adult rats treated neonatally with streptozotocin. Endocrinology. 1990 Apr;126(4):1842–1848. doi: 10.1210/endo-126-4-1842. [DOI] [PubMed] [Google Scholar]
  39. Zawalich W. S., Zawalich K. C., Shulman G. I., Rossetti L. Chronic in vivo hyperglycemia impairs phosphoinositide hydrolysis and insulin release in isolated perifused rat islets. Endocrinology. 1990 Jan;126(1):253–260. doi: 10.1210/endo-126-1-253. [DOI] [PubMed] [Google Scholar]

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

RESOURCES