Troglitazone increases the number of small adipocytes without the change of white adipose tissue mass in obese Zucker rats (original) (raw)

Abstract

Troglitazone (CS-045) is one of the thiazolidinediones that activate the peroxisome proliferator-activated receptor gamma (PPARgamma), which is expressed primarily in adipose tissues. To elucidate the mechanism by which troglitazone relieves insulin resistance in vivo, we studied its effects on the white adipose tissues of an obese animal model (obese Zucker rat). Administration of troglitazone for 15 d normalized mild hyperglycemia and marked hyperinsulinemia in these rats. Plasma triglyceride level was decreased by troglitazone in both obese and lean rats. Troglitazone did not change the total weight of white adipose tissues but increased the number of small adipocytes (< 2,500 micron2) approximately fourfold in both retroperitoneal and subcutaneous adipose tissues of obese rats. It also decreased the number of large adipocytes (> 5,000 micron2) by approximately 50%. In fact, the percentage of apoptotic nuclei was approximately 2.5-fold higher in the troglitazone-treated retroperitoneal white adipose tissue than control. Concomitantly, troglitazone normalized the expression levels of TNF-alpha which were elevated by 2- and 1.4-fold in the retroperitoneal and mesenteric white adipose tissues of the obese rats, respectively. Troglitazone also caused a dramatic decrease in the expression levels of leptin, which were increased by 4-10-fold in the white adipose tissues of obese rats. These results suggest that the primary action of troglitazone may be to increase the number of small adipocytes in white adipose tissues, presumably via PPARgamma. The increased number of small adipocytes and the decreased number of large adipocytes in white adipose tissues of troglitazone-treated obese rats appear to be an important mechanism by which increased expression levels of TNF-alpha and higher levels of plasma lipids are normalized, leading to alleviation of insulin resistance.

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  1. Abbott W. G., Foley J. E. Comparison of body composition, adipocyte size, and glucose and insulin concentrations in Pima Indian and Caucasian children. Metabolism. 1987 Jun;36(6):576–579. doi: 10.1016/0026-0495(87)90170-3. [DOI] [PubMed] [Google Scholar]
  2. Apweiler R., Freund P. Development of glucose intolerance in obese (fa/fa) Zucker rats. Horm Metab Res. 1993 Oct;25(10):521–524. doi: 10.1055/s-2007-1002165. [DOI] [PubMed] [Google Scholar]
  3. Berger J., Bailey P., Biswas C., Cullinan C. A., Doebber T. W., Hayes N. S., Saperstein R., Smith R. G., Leibowitz M. D. Thiazolidinediones produce a conformational change in peroxisomal proliferator-activated receptor-gamma: binding and activation correlate with antidiabetic actions in db/db mice. Endocrinology. 1996 Oct;137(10):4189–4195. doi: 10.1210/endo.137.10.8828476. [DOI] [PubMed] [Google Scholar]
  4. Boden G. Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes. 1997 Jan;46(1):3–10. [PubMed] [Google Scholar]
  5. Braissant O., Foufelle F., Scotto C., Dauça M., Wahli W. Differential expression of peroxisome proliferator-activated receptors (PPARs): tissue distribution of PPAR-alpha, -beta, and -gamma in the adult rat. Endocrinology. 1996 Jan;137(1):354–366. doi: 10.1210/endo.137.1.8536636. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Cleary M. P., Brasel J. A., Greenwood M. R. Developmental changes in thymidine kinase, DNA, and fat cellularity in Zucker rats. Am J Physiol. 1979 May;236(5):E508–E513. doi: 10.1152/ajpendo.1979.236.5.E508. [DOI] [PubMed] [Google Scholar]
  8. Cohen B., Novick D., Rubinstein M. Modulation of insulin activities by leptin. Science. 1996 Nov 15;274(5290):1185–1188. doi: 10.1126/science.274.5290.1185. [DOI] [PubMed] [Google Scholar]
  9. Forman B. M., Tontonoz P., Chen J., Brun R. P., Spiegelman B. M., Evans R. M. 15-Deoxy-delta 12, 14-prostaglandin J2 is a ligand for the adipocyte determination factor PPAR gamma. Cell. 1995 Dec 1;83(5):803–812. doi: 10.1016/0092-8674(95)90193-0. [DOI] [PubMed] [Google Scholar]
  10. Fujiwara T., Okuno A., Yoshioka S., Horikoshi H. Suppression of hepatic gluconeogenesis in long-term Troglitazone treated diabetic KK and C57BL/KsJ-db/db mice. Metabolism. 1995 Apr;44(4):486–490. doi: 10.1016/0026-0495(95)90056-x. [DOI] [PubMed] [Google Scholar]
  11. Fujiwara T., Yoshioka S., Yoshioka T., Ushiyama I., Horikoshi H. Characterization of new oral antidiabetic agent CS-045. Studies in KK and ob/ob mice and Zucker fatty rats. Diabetes. 1988 Nov;37(11):1549–1558. doi: 10.2337/diab.37.11.1549. [DOI] [PubMed] [Google Scholar]
  12. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hofmann C., Lorenz K., Braithwaite S. S., Colca J. R., Palazuk B. J., Hotamisligil G. S., Spiegelman B. M. Altered gene expression for tumor necrosis factor-alpha and its receptors during drug and dietary modulation of insulin resistance. Endocrinology. 1994 Jan;134(1):264–270. doi: 10.1210/endo.134.1.8275942. [DOI] [PubMed] [Google Scholar]
  14. Hotamisligil G. S., Budavari A., Murray D., Spiegelman B. M. Reduced tyrosine kinase activity of the insulin receptor in obesity-diabetes. Central role of tumor necrosis factor-alpha. J Clin Invest. 1994 Oct;94(4):1543–1549. doi: 10.1172/JCI117495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hotamisligil G. S., Johnson R. S., Distel R. J., Ellis R., Papaioannou V. E., Spiegelman B. M. Uncoupling of obesity from insulin resistance through a targeted mutation in aP2, the adipocyte fatty acid binding protein. Science. 1996 Nov 22;274(5291):1377–1379. doi: 10.1126/science.274.5291.1377. [DOI] [PubMed] [Google Scholar]
  16. Hotamisligil G. S., Murray D. L., Choy L. N., Spiegelman B. M. Tumor necrosis factor alpha inhibits signaling from the insulin receptor. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4854–4858. doi: 10.1073/pnas.91.11.4854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hotamisligil G. S., Peraldi P., Budavari A., Ellis R., White M. F., Spiegelman B. M. IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in TNF-alpha- and obesity-induced insulin resistance. Science. 1996 Feb 2;271(5249):665–668. doi: 10.1126/science.271.5249.665. [DOI] [PubMed] [Google Scholar]
  18. Hotamisligil G. S., Shargill N. S., Spiegelman B. M. Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science. 1993 Jan 1;259(5091):87–91. doi: 10.1126/science.7678183. [DOI] [PubMed] [Google Scholar]
  19. Iwamoto Y., Kuzuya T., Matsuda A., Awata T., Kumakura S., Inooka G., Shiraishi I. Effect of new oral antidiabetic agent CS-045 on glucose tolerance and insulin secretion in patients with NIDDM. Diabetes Care. 1991 Nov;14(11):1083–1086. doi: 10.2337/diacare.14.11.1083. [DOI] [PubMed] [Google Scholar]
  20. Kallen C. B., Lazar M. A. Antidiabetic thiazolidinediones inhibit leptin (ob) gene expression in 3T3-L1 adipocytes. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5793–5796. doi: 10.1073/pnas.93.12.5793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kellerer M., Kroder G., Tippmer S., Berti L., Kiehn R., Mosthaf L., Häring H. Troglitazone prevents glucose-induced insulin resistance of insulin receptor in rat-1 fibroblasts. Diabetes. 1994 Mar;43(3):447–453. doi: 10.2337/diab.43.3.447. [DOI] [PubMed] [Google Scholar]
  22. Kliewer S. A., Lenhard J. M., Willson T. M., Patel I., Morris D. C., Lehmann J. M. A prostaglandin J2 metabolite binds peroxisome proliferator-activated receptor gamma and promotes adipocyte differentiation. Cell. 1995 Dec 1;83(5):813–819. doi: 10.1016/0092-8674(95)90194-9. [DOI] [PubMed] [Google Scholar]
  23. Kliewer S. A., Umesono K., Noonan D. J., Heyman R. A., Evans R. M. Convergence of 9-cis retinoic acid and peroxisome proliferator signalling pathways through heterodimer formation of their receptors. Nature. 1992 Aug 27;358(6389):771–774. doi: 10.1038/358771a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kumar S., Boulton A. J., Beck-Nielsen H., Berthezene F., Muggeo M., Persson B., Spinas G. A., Donoghue S., Lettis S., Stewart-Long P. Troglitazone, an insulin action enhancer, improves metabolic control in NIDDM patients. Troglitazone Study Group. Diabetologia. 1996 Jun;39(6):701–709. doi: 10.1007/BF00418542. [DOI] [PubMed] [Google Scholar]
  25. Lehmann J. M., Moore L. B., Smith-Oliver T. A., Wilkison W. O., Willson T. M., Kliewer S. A. An antidiabetic thiazolidinedione is a high affinity ligand for peroxisome proliferator-activated receptor gamma (PPAR gamma). J Biol Chem. 1995 Jun 2;270(22):12953–12956. doi: 10.1074/jbc.270.22.12953. [DOI] [PubMed] [Google Scholar]
  26. Maffei M., Fei H., Lee G. H., Dani C., Leroy P., Zhang Y., Proenca R., Negrel R., Ailhaud G., Friedman J. M. Increased expression in adipocytes of ob RNA in mice with lesions of the hypothalamus and with mutations at the db locus. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6957–6960. doi: 10.1073/pnas.92.15.6957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Montague C. T., Prins J. B., Sanders L., Digby J. E., O'Rahilly S. Depot- and sex-specific differences in human leptin mRNA expression: implications for the control of regional fat distribution. Diabetes. 1997 Mar;46(3):342–347. doi: 10.2337/diab.46.3.342. [DOI] [PubMed] [Google Scholar]
  28. Müller G., Ertl J., Gerl M., Preibisch G. Leptin impairs metabolic actions of insulin in isolated rat adipocytes. J Biol Chem. 1997 Apr 18;272(16):10585–10593. doi: 10.1074/jbc.272.16.10585. [DOI] [PubMed] [Google Scholar]
  29. Ohsumi J., Sakakibara S., Yamaguchi J., Miyadai K., Yoshioka S., Fujiwara T., Horikoshi H., Serizawa N. Troglitazone prevents the inhibitory effects of inflammatory cytokines on insulin-induced adipocyte differentiation in 3T3-L1 cells. Endocrinology. 1994 Nov;135(5):2279–2282. doi: 10.1210/endo.135.5.7956951. [DOI] [PubMed] [Google Scholar]
  30. Olefsky J. M. The effects of spontaneous obesity on insulin binding, glucose transport, and glucose oxidation of isolated rat adipocytes. J Clin Invest. 1976 Apr;57(4):842–851. doi: 10.1172/JCI108360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pedersen O., Kahn C. R., Kahn B. B. Divergent regulation of the Glut 1 and Glut 4 glucose transporters in isolated adipocytes from Zucker rats. J Clin Invest. 1992 Jun;89(6):1964–1973. doi: 10.1172/JCI115804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pennica D., Hayflick J. S., Bringman T. S., Palladino M. A., Goeddel D. V. Cloning and expression in Escherichia coli of the cDNA for murine tumor necrosis factor. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6060–6064. doi: 10.1073/pnas.82.18.6060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Prins J. B., Walker N. I., Winterford C. M., Cameron D. P. Apoptosis of human adipocytes in vitro. Biochem Biophys Res Commun. 1994 Jun 15;201(2):500–507. doi: 10.1006/bbrc.1994.1730. [DOI] [PubMed] [Google Scholar]
  34. RANDLE P. J., GARLAND P. B., HALES C. N., NEWSHOLME E. A. The glucose fatty-acid cycle. Its role in insulin sensitivity and the metabolic disturbances of diabetes mellitus. Lancet. 1963 Apr 13;1(7285):785–789. doi: 10.1016/s0140-6736(63)91500-9. [DOI] [PubMed] [Google Scholar]
  35. Saltiel A. R., Olefsky J. M. Thiazolidinediones in the treatment of insulin resistance and type II diabetes. Diabetes. 1996 Dec;45(12):1661–1669. doi: 10.2337/diab.45.12.1661. [DOI] [PubMed] [Google Scholar]
  36. Schoonjans K., Staels B., Auwerx J. The peroxisome proliferator activated receptors (PPARS) and their effects on lipid metabolism and adipocyte differentiation. Biochim Biophys Acta. 1996 Jul 26;1302(2):93–109. doi: 10.1016/0005-2760(96)00066-5. [DOI] [PubMed] [Google Scholar]
  37. Sears I. B., MacGinnitie M. A., Kovacs L. G., Graves R. A. Differentiation-dependent expression of the brown adipocyte uncoupling protein gene: regulation by peroxisome proliferator-activated receptor gamma. Mol Cell Biol. 1996 Jul;16(7):3410–3419. doi: 10.1128/mcb.16.7.3410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Segal K. R., Landt M., Klein S. Relationship between insulin sensitivity and plasma leptin concentration in lean and obese men. Diabetes. 1996 Jul;45(7):988–991. doi: 10.2337/diab.45.7.988. [DOI] [PubMed] [Google Scholar]
  39. Sreenan S., Sturis J., Pugh W., Burant C. F., Polonsky K. S. Prevention of hyperglycemia in the Zucker diabetic fatty rat by treatment with metformin or troglitazone. Am J Physiol. 1996 Oct;271(4 Pt 1):E742–E747. doi: 10.1152/ajpendo.1996.271.4.E742. [DOI] [PubMed] [Google Scholar]
  40. Stephens J. M., Pekala P. H. Transcriptional repression of the GLUT4 and C/EBP genes in 3T3-L1 adipocytes by tumor necrosis factor-alpha. J Biol Chem. 1991 Nov 15;266(32):21839–21845. [PubMed] [Google Scholar]
  41. Stevenson R. W., Hutson N. J., Krupp M. N., Volkmann R. A., Holland G. F., Eggler J. F., Clark D. A., McPherson R. K., Hall K. L., Danbury B. H. Actions of novel antidiabetic agent englitazone in hyperglycemic hyperinsulinemic ob/ob mice. Diabetes. 1990 Oct;39(10):1218–1227. doi: 10.2337/diab.39.10.1218. [DOI] [PubMed] [Google Scholar]
  42. Suter S. L., Nolan J. J., Wallace P., Gumbiner B., Olefsky J. M. Metabolic effects of new oral hypoglycemic agent CS-045 in NIDDM subjects. Diabetes Care. 1992 Feb;15(2):193–203. doi: 10.2337/diacare.15.2.193. [DOI] [PubMed] [Google Scholar]
  43. Szalkowski D., White-Carrington S., Berger J., Zhang B. Antidiabetic thiazolidinediones block the inhibitory effect of tumor necrosis factor-alpha on differentiation, insulin-stimulated glucose uptake, and gene expression in 3T3-L1 cells. Endocrinology. 1995 Apr;136(4):1474–1481. doi: 10.1210/endo.136.4.7895657. [DOI] [PubMed] [Google Scholar]
  44. Taylor S. I., Barr V., Reitman M. Does leptin contribute to diabetes caused by obesity? Science. 1996 Nov 15;274(5290):1151–1152. doi: 10.1126/science.274.5290.1151. [DOI] [PubMed] [Google Scholar]
  45. Tontonoz P., Hu E., Graves R. A., Budavari A. I., Spiegelman B. M. mPPAR gamma 2: tissue-specific regulator of an adipocyte enhancer. Genes Dev. 1994 May 15;8(10):1224–1234. doi: 10.1101/gad.8.10.1224. [DOI] [PubMed] [Google Scholar]
  46. Tontonoz P., Hu E., Spiegelman B. M. Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell. 1994 Dec 30;79(7):1147–1156. doi: 10.1016/0092-8674(94)90006-x. [DOI] [PubMed] [Google Scholar]
  47. Wang Y., Kuropatwinski K. K., White D. W., Hawley T. S., Hawley R. G., Tartaglia L. A., Baumann H. Leptin receptor action in hepatic cells. J Biol Chem. 1997 Jun 27;272(26):16216–16223. doi: 10.1074/jbc.272.26.16216. [DOI] [PubMed] [Google Scholar]
  48. Willson T. M., Cobb J. E., Cowan D. J., Wiethe R. W., Correa I. D., Prakash S. R., Beck K. D., Moore L. B., Kliewer S. A., Lehmann J. M. The structure-activity relationship between peroxisome proliferator-activated receptor gamma agonism and the antihyperglycemic activity of thiazolidinediones. J Med Chem. 1996 Feb 2;39(3):665–668. doi: 10.1021/jm950395a. [DOI] [PubMed] [Google Scholar]
  49. Zhang B., Graziano M. P., Doebber T. W., Leibowitz M. D., White-Carrington S., Szalkowski D. M., Hey P. J., Wu M., Cullinan C. A., Bailey P. Down-regulation of the expression of the obese gene by an antidiabetic thiazolidinedione in Zucker diabetic fatty rats and db/db mice. J Biol Chem. 1996 Apr 19;271(16):9455–9459. doi: 10.1074/jbc.271.16.9455. [DOI] [PubMed] [Google Scholar]
  50. Zhang Y., Proenca R., Maffei M., Barone M., Leopold L., Friedman J. M. Positional cloning of the mouse obese gene and its human homologue. Nature. 1994 Dec 1;372(6505):425–432. doi: 10.1038/372425a0. [DOI] [PubMed] [Google Scholar]