Phloretin inhibits Na+ and K+ uptake in alveolar epithelial type cells via a reduction of cellular atp content
Abstract
Bipolar organic compound phloretin has a marked effect on transport and celular metabolism of glucose, uptake of non-metabolized glucose analogs and epithelial cell growth. We have studied effect of phloretin on Na+ and Rb+ uptake in rat alveolar epithelial type cells. Type cells were isolated by elastase digestion, purified by a differential adherential technique and than cultured on plastic in Dulbeccos medium with 10% fetal bovin serum. 22Na+ and 86Rb+ were used as trasers for Na+ and К+ accumulation in the presence or absence of phloretin (0–250 µM). The ATP content was determined by the adapted fireify luciferase assay.
Incubation of type cells with various concentration of phloretin leads to inhibition of Na and Rb uptake and a reduction of cellular ATP content. The effect is present in the presence and absence of external glucose and maximum inhibition was detected at a concentration of 250 µM. Sodium uptake was decreased from 46.0±2.7 in control up to 5.5+0.8 nmol/mg protein/5 min in the presence of 250 mM of phloretin and rubidium/potassium uptake was decreased from 35.3+2.8 to 1.8 nmol/mg protein /15 min. At the same concentration of drug cellular ATP content was reduced from 0 .9 6+ 0 .1 6 to 0 .0 3± 0 .0 2 nmol/mg protein. The results indicate that phloretin induces a reduction in cellular ATP content which lead to decrease in Na+ and К+ uptake in type 11 epithelial cells.
About the Authors
G. A. Skrycibin
НИИ пульмонологии М3 РФ
Russian Federation
Russian Federation
S. N. Orlov
Исследовательский Центр Университета
Canada
Canada
G. G. Ryazhskcy
НИИ пульмонологии М3 РФ
Russian Federation
Russian Federation
N. I. Pokudin
НИИ пульмонологии М3 РФ
Russian Federation
Russian Federation
I. Bertjem
Исследовательский Центр Университета
Canada
Canada
A. G. Chuchalin
НИИ пульмонологии М3 РФ
Russian Federation
Russian Federation
References
1. Basset G., Crone С., Saumon G. Significance of active ion transport in transalveolar water absorption: a study on isolated rat lung // J. Physiol. Lond. — Vol. 384. — P. 311—324.
2. Basset G., Crone C., Saumon G. Fluid absorption by rat lung in situ: pathways for sodium entry in the luminal membrane of alveolar epithelium // Ibid. — P. 325—345.
3. Basset G., Saumon G., Bouchonnet F., Crone C. Apical sodium transport in pulmonary epithelium in situ // Biochim. Biophys. Acta. — 1988. — Vol. 942, № 1. — P. l1—18.
4. Bland R.D., Boyd C.A.R. Cation transport in lung epithelial cells derived from fetal, newborn and adult rabbits // J. Appl. Physiol. — 1986. — Vol. 61, № 2. — P. 507—515.
5. Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding // Anal. Biochem. — 1976. — Vol. 72. — P. 248—254.
6. Brown S.E.S., Kim K.J., Goodman B.E., Wells J.R., Crandall E.D. Sodium-amino acid cotransport by type II alveolar epithelial cells // J. Appl. Physiol. — 1985. — Vol. 59. — Р. 1616—1622.
7. Brown S.E.S., Heming T.A., Benedict C.R., Bidani A. ATP-sensitive Na -H antiport in type II alveolar epithelial cells // Am. J. Physiol. — 1991. — Vol. 261. — P. C954—C963.
8. Clerici C., Soler P., Saumon G. Sodium dependent phosphate and alanine transport but sodium-independent hexose transport in type II alveolar epithelial cells in primary culture // Biochim. Biophys. Acta. — 1991. — Vol. 1063. — P. 27—35.
9. De Jonge P.C., Wieringa Т., Van Put ten J.P., Krans H.M., Van Dam K. Phloretin - an uncoupler and an inhibitor of mitochondrial oxidative phosphorilation // Ibid. — 1983. — Vol. 772. — P. 219—225.
10. Delamere N., Coca-Prados М., Aggarwal S. Studies on regulation of the ascorbic acid transporter in a cell line derived from rabbit non-pigmented ciliary epithelium // Ibid. — 1993. — Vol. l149. — P. 102—108.
11. Dobbs L.G., Gonzalez R., Williams M.C. An improved method for isolating type II ceils in high yield and purity // Am. Rev. Respir. Dis. — 1986. — Vol. 134. — P. 1268—1275.
12. Edelson J.D., Shannon J.M., Mason R.J. Alkaline phosphatase: a marker of alveolar type II cells differentiation // Ibid. — 1988. — Vol. 138. — P. 1268—1275.
13. Feng Z.-P., Clark R.B., Berthiaume Y. Identification of nonselective cation channels in cultured adult rat alveolar type II cells // Am. J. Respir. Cell Mol. Biol. — 1993. — Vol. 9. — P. 248—254.
14. Fisher A.B., Furia L.t Berman H. Metabolism of rat granular pneumocytes isolated in primary culture // J. Appl. Physiol. — 1980. — Vol. 49. — P. 743—750.
15. Helbig H., Korbmocher C., Wohlfarth J., Berweck S., Kuhner D., Wiederholt M. Electrogenic Na+-ascorbate cotransport in cultured bovine pigmented ciliary epithelial cells // Am. J. Physiol. — 1989. — Vol. 256. — P. C44—C49.
16. Kemp P.J., Boyd C.A. Pathways for glucose transport in type II pneumocytes freshly isolated from adult guinea pig lung // Ibid. — 1992. — Vol. 263. — P. L612—L616.
17. Kemp P.J., Roberts G.C., Boyd C.A.R. Identification and properties of pathways for K+ transport in guinea-pig and rat alveolar epithelial type II cells // J. Physiol. Lond. — 1994. - Vol. 476. - P. 79—88.
18. Kerr J.S., Reicherter J., Fisher A.B. 2-Deoxy-D-glucose uptake by rat granular pneumocytes in primary culture // Am. J. Physiol. — 1982. — Vol. 243. — P. C14—C19.
19. Kimmich G.A., Randles J. A Na-independent, phloretin-sensitive monosacharide transport system in isolated intestinal epithelial cells // J. Membr. Biol. — 1975. — Vol. 23, № 1. — P. 57—76.
20. Kimmich G.A., Carter-Su C., Randles J. Energetic of Na-dependent, sugar transport by isolated intestinal epithelial cells: evidence for a major role for membrane potentials // Am. J. Physiol. — 1977. — Vol. 233, № 5. — P. E357—E362.
21. LaCagnin L.B., Bowman L., Ma J.Y.C., Miles P.R. Metabolic changes in alveolar type II cells after exposure to hydrogen peroxide // Ibid. — 1990. — Vol. 259. — P. L57—L65.
22. Lumban R.L., Crandall E.D. Na+-HC03 symport modulates intracellular pH in alveolar epithelial cells // Ibid. — 1991. — Vol. 260. — P. L555—L561.
23. Matalon S. Mechanism and requlation of ion transport in abuld and mammalian alveolar type II pneumocytes // Ibid. — 1991. — Vol. 261. - P. C727—C738.
24. Matalon S., Bauer M.L., Benos D.J., Kleyman T.R., Lin C., Cragoe E.J., O’Brodovich H. Fetal lung epithelial cells contain two populations of amiloride-sensitive Na channels // Ibid. — 1993. — Vol. 264. — P. L357—L364.
25. Matalon S., Bridges R., Benos D.J. Amiloride-inhabitable Na conductive pathways in alveolar type II pneumocytes // Ibid. — 1991. - Vol. 260. — P. L90—L96.
26. Matthay M.A., Berthiaume Y., Staub N.C. Long-term clearance of liquid and protein from the lung of unasthezed sheep // J. Appl. Physiol. — 1985. — Vol. 59. — P. 928—934.
27. Michell D.B., Santone K.S., Acosta D. Evaluation of cytoxicity in cultured cells by enzyme leakage // J. Tissue Culture Meth. — 1980. — Vol. 6. — P. 113—116.
28. Mullin J.M., Kofeldt L.M., Russo L.M., Hagee M.M., Dantzig A.H. Basolateral 3-O-methyl glucose transport by cultured kidney (LLC-PK1) epihtelial cells // Am. J. Physiol. — 1992. — Vol. 262, № 3. - Pt 2. — P. F480—F487.
29. Nord E.P., Brown S.E.S., Crandall E.D. Characterization of Na+- H+ antiport in type II alveolar epithelial cells // Ibid. — 1987. — Vol. 252. — P. C490—C498.
30. O’Brodouich H.,Rafii B., Post M. Bioelectric properties of fetal alveolar epithelial monolayers // Ibid. — 1990. — Vol. 258. — P. L201—L206.
31. O’Brodovich H., Hannam V., Seear M., Mullen J.B.M. Amiloride impairs lung water clearance in newborn guinea pigs // J. Appl. Physiol. — 1990. — Vol. 68. — P. 1758—1762.
32. Oelberg D.G., Xuy F., Shabarek F. Sodium-coupled transport of glucose by plasma membranes of type II pneumocytes // Biochim. Biophys. Acta. — 1994. — Vol. 1194. — P. 92—98.
33. Russo R.M., Lubman R.L., Crandall E.D. Evidence for amiloridesensitive sodium channels in alveolar epithelial cells // Am. J. Physiol. — 1992. — Vol. 262. — P. L405—L411.
34. Lubman R.L., Crandall E.D. Polarized distribution of Na+-H+ antiport activity in rat alveolar epithelial cells // Ibid. — 1994. — Vol. 266. — P. L138—L147.
35. Saumon G., Basset G. Electrolyte and fluid transport across the mature alveolar epithelium // J. Appl. Physiol. — 1993. — Vol. 74, № 1. — P. l—15.
36. Grinstein S. et al. Mechanisms of regulation of Na-H echanger // J. Membr. Biol. — 1986. — Vol. 90. — P. l—12.
37. Smedira N., Gates L., Hastings R.t Jayr C., SakumaT., Pittet J.F., Matthay M.A. Alveolar and lung liquid clearance in anestathized rabbits // J. Appl. Physiol. — 1991. — Vol. 70. — P. 1827—1835.
38. Verkman A.S., Solomon A.K. A stepwise merchanism for the permeation of phloretin through a lipid bilayer // J. Gen. Physiol. — 1982. — Vol. 80, № 4. — P. 557—581.
39. Wangensteen D., Barlett M. D-and L-glucose transport across the pulmonary epithelium // J. Appl. Physiol. — 1984. — Vol. 57, № 6. — P. 1722—1730.
40. Waters B., Thakar J., Lapierre Y. Erythrocyte lithium transport variables as a marker for manic-depressive // Neuropsychobiology. — 1983. — Vol. 9, № 2. — P. 94—98.
41. Yue G., Shoemaker R.L., Matalon S. Regulation of low-amilorideaffinity sodium channels in alveolar type II cells // Am. J. Physiol. — 1994. — Vol. 267. — P. L94—L100.
42. Aiton J.F., Brown C.D., Ogden P., Simmons N.L. K+ transport in “tight”epithelial monolayers of MDCK cells // J. Membr. Biol. — 1982. — Vol. 65. — № 1—2. — P. 99—109.
43. Aiton J.F., Chipperfield A.R., Lamp J.F., Ogden P., Simmons N.L. Occurrence of passive furosemide-sensitive transmembrane potessium transport in cultured cells // Biochim. Biophys. Acta. — 1981. — Vol. 646, № 3. — P. 389—398.
44. O’Grady S.M., Palfrey H.C., Field M. Characteristics and functions of Na-K-2C1 cotransport in epithelial tissued // Am. J. Physiol. — 1987. — Vol. 253. — P. C177—C192.
45. Jorgensen P.L. Mechanism of the Na+-K+-pump. Protein structure and conformations of pure (Na+-K+)-ATPase // Biochim. Biophys. Acta. — 1982. — Vol. 694, № 1. — P. 27—68.
46. Chipperfield A.R. The Na+-K+-CI' cotranspost system // Clin. Sci. — 1986. — Vol. 71, № 5. — P. 465—476.
Review
For citations:
Skrycibin G.A., Orlov S.N., Ryazhskcy G.G., Pokudin N.I., Bertjem I., Chuchalin A.G. Phloretin inhibits Na+ and K+ uptake in alveolar epithelial type cells via a reduction of cellular atp content. PULMONOLOGIYA. 1996;(3):58-64. (In Russ.)
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