Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Effect of protein kinase C modulators on the leucocyte Na+/H+ antiport in Type 1 (insulin-dependent) diabetic subjects with albuminuria


It is uncertain why only one third of Type 1 (insulin-dependent) diabetic patients develop nephropathy. One suggestion is the inheritance of a predisposition to essential hypertension. We have previously found elevated Na+/H+ antiport activity and a raised intracellular pH in leucocytes from hypertensive and Type 1 diabetic subjects with albuminuria using a novel double ionophore fluorimetric technique. These changes are not found in Type 1 diabetic subjects without albuminuria. We wished to test the effect of a protein kinase C inhibitor staurosporine (100 nmol/l) on the elevated antiport activity, and the degree of stimulation achieved by exogenous diacyl glycerol. Raised leucocyte Na+/H+ antiport activity of Type 1 diabetic subjects with albuminuria (73.8±17.2 mmol·l−1·min−1) was restored to normal levels with staurosporine (54.9±17.9 mmol·l−1·min−1, p<0.001). The leucocyte Na+/H+ antiport activity of diabetic subjects without albuminuria fell significantly also with staurosporine but to a lesser extent (57.3±11.6 to 50.0±12.8 mmol/l, p<0.003). In contrast, leucocytes from normal control subjects showed no change in antiport activity with staurosporine (54.3±8.5 to 52.6±10.4 mmol ·1−1·min−1). Dioctanoyl glycerol stimulated the leucocyte Na+/H+ antiport in normal subjects and diabetic patients without albuminuria, with significantly less stimulation in diabetic patients with albuminuria. We conclude that reversal by staurosporine of the elevated Na+/H+ antiport activity in Type 1 diabetic subjects with albuminuria could indicate a role for protein kinase C in activating the antiport. This hypothesis is supported by the reduced stimulation of the antiport by dioctanoyl glycerol in this group of patients.


  1. 1.

    Krolewski AS, Warram JH, Christlieb AR, Busick EJ, Kahn CR (1985) The changing natural history of nephropathy in type 1 diabetes. Am J Med 78: 785–794

  2. 2.

    Feldt-Rasmussen B, Mathiesen ER, Deckert T (1986) Effect of two years of strict metabolic control on progression of incipient nephropathy in insulin-dependent diabetes. Lancet II: 1300–1304

  3. 3.

    Nørgaard K, Strom B, Graae M, Feldt-Rasmussen B (1988) Microalbuminuria and retinal changes in children with Type 1 (insulin-dependent) diabetes mellitus are related to long-term poor metabolic control. Diabetologia 31: 527 Abstract

  4. 4.

    Seaquist ER, Goetz FC, Rich S, Barbosa J (1987) Concordance for nephropathy in diabetic siblings: evidence of genetic susceptibility to diabetic kidney disease. Diabetes 36 [Suppl]: 105A

  5. 5.

    Krolewski AS, Canessa M, Warram JH, Laffel LMB, Christlieb AR, Knowler WC, Rand LI (1988) Predisposition to hypertension and susceptibility to renal disease in insulin-dependent diabetes mellitus. N Engl J Med 318: 140–145

  6. 6.

    Viberti GC, Keen H, Wiseman MJ (1987) Raised arterial pressure in parents of proteinuric insulin dependent diabetics. Br Med J 295: 515–517

  7. 7.

    Hilton PJ (1986) Cellular sodium transport in essential hypertension. N Engl J Med 314: 222–229

  8. 8.

    Canessa M, Adragna N, Solomon HS, Connolly TM, Tosteson DC (1980) Increased sodium-lithium countertransport in red cells of patients with essential hypertension. N Engl J Med 302: 772–776

  9. 9.

    Hasstedt SJ, Wu LL, Ash KO, Kuida H, Williams RR (1988) Hypertension and sodium-lithium countertransport in Utah pedigrees: evidence for major-locus inheritance. Am J Hum Genet 43: 14–22

  10. 10.

    Mangili R, Bending JJ, Scott G, Li LK, Gupta A, Viberti GC (1988) Increased sodium-lithium countertransport activity in red cells of patients with insulin-dependent diabetes and nephropathy. N Engl J Med 318: 146–150

  11. 11.

    Mahnensmith RL, Aronson PS (1985) The plasma membrane sodium-hydrogen exchanger and its role in physiological and pathophysiological processes. Circ Res 56: 773–788

  12. 12.

    Livne A, Balfe JW, Veitch R, Marquez-Julio A, Grinstein S, Rothstein A (1987) Increased platelet Na+-H+ exchange rate in essential hypertension: application of a novel test. Lancet II: 533–536

  13. 13.

    Ng LL, Harker M, Abel ED (1988) Mechanisms of leucocyte sodium influx in essential hypertension. Clin Sci 75: 521–526

  14. 14.

    Ng LL, Dudley C, Bomford J, Hawley D (1989) Leucocyte intracellular pH and Na+/H+ antiport activity in human hypertension. J Hypertens 7: 471–475

  15. 15.

    Postnov YV, Kravtsov GM, Orlov SN, Pokudin NI, Postnov IY, Kotelevtsev YV (1988) Effect of protein kinase C activation on cytoskeleton and cation transport in human erythrocytes. Reproduction of some membrane abnormalities revealed in essential hypertension. Hypertension 12: 267–273

  16. 16.

    Feig PU, D'Occhio MA, Boylan JW (1987) Lymphocyte membrane sodium-proton exchange in spontaneously hypertensive rats. Hypertension 9: 282–288

  17. 17.

    Berk BC, Vallega G, Muslin AJ, Gordon HM, Canessa M, Alexander RW (1989) Spontaneously hypertensive rat vascular smooth muscle cells in culture exhibit increased growth and Na+/H+ exchange. J Clin Invest 83: 822–829

  18. 18.

    Uehara Y, Ishii M, Ishimitsu T, Sugimoto T (1988) Enhanced phospholipase C activity in the vascular wall of spontaneously hypertensive rats. Hypertension 11: 28–33

  19. 19.

    Ng LL, Simmons D, Frighi V, Garrido MC, Bomford J, Hockaday TDR (1990) Leucocyte Na/H antiport activity in Type 1 (insulin-dependent) diabetic patients with nephropathy. Diabetologia 33: (in press)

  20. 20.

    Tamaoki T, Nomoto H, Takahashi I, Kato Y, Morimoto M, Tomita F (1986) Staurosporine, a potent inhibitor of phospholipid/Ca+ + dependent protein kinase. Biochem Biophys Res Commun 135: 397–402

  21. 21.

    Flavell HJ, Capps NE, Bottomley S (1988) A manual immunoturbidimetric method for low concentrations of albumin adapted to the IL “Multistat 3 Plus”. Clin Chem 34: 1506

  22. 22.

    Gatling WG, Knight C, Mullee MA, Hill RD (1988) Microalbuminuria in diabetes; A population study of the prevalence and an assessment of three screening tests. Diabetic Med 5: 343–347

  23. 23.

    Hanning I, Home PD, Alberti KGMM (1985) Measurement of free insulin concentrations: the influence of the timing of extraction of insulin antibodies. Diabetologia 28: 831–835

  24. 24.

    Baron DN, Ahmed SA (1969) Intracellular concentration of water and of the principal electrolytes determined by analysis of isolated human leucocytes. Clin Sci 37: 205–219

  25. 25.

    Grinstein S, Cohen S, Rothstein A (1984) Cytoplasmic pH regulation in thymic lymphocytes by an amiloride-sensitive Na/H antiport. J Gen Physiol 83: 341–370

  26. 26.

    Ng LL, Dudley C (1989) Intracellular pH clamping of human leucocytes: a technique for determination of cellular buffering power and Na+/H+ antiport characteristics. Clin Sci 77: 411–423

  27. 27.

    Ng LL, Bomford J (1989) Altered stoichiometry of the human leucocyte Na+/H+ antiport with decreasing pH. Biochem J 259: 311–314

  28. 28.

    Roos A, Boron WF (1981) Intracellular pH. Physiol Rev 61: 296–434

  29. 29.

    Boyarsky G, Ganz MB, Sterzel RB, Boron WF (1988) pH regulation in single glomerular mesangial cells I. Acid extrusion in the absence and presence of HCO3 . Am J Physiol 255: C844–856

  30. 30.

    Colquhoun D (1971) Fitting curves: the relationship between two variables. In: Lectures on biostatistics. Clarendon Press, Oxford, pp 262–265

  31. 31.

    Lee T-S, MacGregor LC, Fluharty SJ, King GL (1989) Differential regulation of protein kinase C and (Na,K)-Adenosine Triphosphatase activities by elevated glucose levels in retinal capillary endothelial cells. J Clin Invest 83: 90–94

  32. 32.

    Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233: 305–312

  33. 33.

    Nishizuka Y (1988) The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature 334: 661–665

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ng, L.L., Simmons, D., Frighi, V. et al. Effect of protein kinase C modulators on the leucocyte Na+/H+ antiport in Type 1 (insulin-dependent) diabetic subjects with albuminuria. Diabetologia 33, 278–284 (1990).

Download citation

Key words

  • Sodium
  • leucocytes
  • amiloride
  • protein kinase
  • Type 1 (insulin-dependent) diabetes