Inhibition of $IL-1{\beta}$ and IL-6 in Osteoblast-Like Cell by Isoflavones Extracted from Sophorae fructus

  • Joo, Seong-Soo (Department of Immunology, College of Pharmacy, Chung-Ang University) ;
  • Kang, Hee-Cheol (Department of Immunology, College of Pharmacy, Chung-Ang University) ;
  • Choi, Min-Won (Department of Immunology, College of Pharmacy, Chung-Ang University) ;
  • Choi, Young-Wook (Department of Immunology, College of Pharmacy, Chung-Ang University) ;
  • Lee, Do-ik (Department of Immunology, College of Pharmacy, Chung-Ang University)
  • 발행 : 2003.11.01

초록

Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When estrogen is reduced in the body, local factors such as IL-1 $\beta$ and IL-6, which are known to be related with bone resorption, are increased and promote osteoclastogenesis, which is responsible for bone resorption. In the present study, we investigated whether glucosidic isoflavones (Isocal, PIII) extracted from Sophorae fructus affect the proliferation of osteoblasts and prevent osteoclastogenesis in vitro by attenuating upstream cytokines such as IL-1$\beta$ and IL-6 in a human osteoblastic cell line (MG-63) and in a primary osteoblastic culture from SD rat femurs. Interestingly, IL-1$\beta$ and IL-6 mRNA were significantly suppressed in osteoblast-like cells treated with 17$\beta$-estradiol (E2) and PIII when compared to positive control (SDB), and this suppression was more effective at $10^{-8}$% than at the highest concentration of $10^{-4}$%. In addition, these were confirmed in protein levels using ELISA assay. In the cell line, the cells showed that E2 was the most effective in osteoblastic proliferation over the whole range of concentration ($10^{-4}%-10^{-12}$%), even though PIII also showed the second greatest effectiveness at $10^{-8}$%. Nitric oxide (NO) was significantly (p<0.05) upregulated in PIII and E2 over the concentration range $10^{-6}% to 10^{-8}$% when compared to SDB, without showing any dose dependency. In bone marrow primary culture, we found by TRAP assay that PIII effectively suppressed osteoclastogenesis next to E2 in comparison with SDB and culture media (control). In conclusion, these results suggest that local bone-resorbing cytokines can be regulated by PIII at lower concentrations and that, therefore, PIII may preferentially induce anti-osteoporosis response by attenuating osteoclastic differentiation and by upregulating NO.

키워드

참고문헌

  1. Adlercreutz, H., Phyto-oestrogens and cancer. Lancet Oncology, 3, 364-373 (2002) https://doi.org/10.1016/S1470-2045(02)00777-5
  2. Amonkar, M. M. and Mody, R., Developing profiles of post-menopausal women being prescribed oestrogen therapy to prevent osteoporosis. J. Commun. Health, 27, 335-350 (2002) https://doi.org/10.1023/A:1019836610275
  3. Armour, K. E. and Ralston, S. H., Estrogen upregulates endothelial constitutive nitric oxide synthase expression in human osteoblast-like cells. Endocrinology, 139, 799-802 (1998) https://doi.org/10.1210/en.139.2.799
  4. Duncan, A. M., Phipps, W. R., and Kurzer, M. S., Phytooestrogen. Best Practice & Research Clinical Endocrinology and Metabolism, 17, 253-271 (2003) https://doi.org/10.1016/S1521-690X(02)00103-3
  5. Eastel, R., Treatment of postmenopausal osteoporosis. N. Engl. J. Med., 338, 736-746 (1998) https://doi.org/10.1056/NEJM199803123381107
  6. Felix, R., Cecchini, M. G., Hofstetter, W., Elford, P. R., Stutzer, A., and Fleishi, H., Impatient of macrophage colony-stimulating factor production and lack of resident bone marrow macrophage in the osteoporotic op/op mouse. J. Bone Miner. Res., 5, 781-789 (1990) https://doi.org/10.1002/jbmr.5650050716
  7. Fujikawa, Y., Sabokbar, A., Neale, D., Itonaga, I., Torisu, T., and Athanasou, A., The effect of macrophage-colony stimulating factor and other humoral factors (interleukin-1, -3, -6, and -11, tumor necrosis factor-$\alpha$, and granulocyte macrophage-colony stimulating factor) on human osteoclast formation from circulating cells. Bone, 28, 261-267 (2001) https://doi.org/10.1016/S8756-3282(00)00453-1
  8. Greenfield, E. M., Bi, Y., and Miyauchi, A., Regulation of osteoclast activity. Life Sicence, 65, 1087-1102 (1999) https://doi.org/10.1016/S0024-3205(99)00156-3
  9. Joo, S. S., Chang, J. K., Park, J. H., Kang, H. C., and Lee, D. I., Immuno activation of lectin-conjugated praecoxin A on IL-6, IL-12 expression. Arch. Pharm. Res., 25, 954-963 (2002) https://doi.org/10.1007/BF02977019
  10. Klein-Nulend, J., Helfrich, M. H., Sterck, J. G., MacPherson, H., Joldersma, M., Ralson, S. H., Semeins, C. M., and Burger, E. H, Nitric oxide response to shear stress by human bone cell culture is endothelial nitric oxide synthase dependant. Biochem. Biophys. Res. Commun., 250, 108-114 (1998) https://doi.org/10.1006/bbrc.1998.9270
  11. Lader, C. S. and Flanagan, A. M., Prostaglandin $E_2$, interleukin $\1{alpha}$, and tumor necrosis factor-$\alpha$ increase human osteoclast formation and bone resorption in vitro. Endocrinology, 139, 3157-3164 (1998) https://doi.org/10.1210/en.139.7.3157
  12. MacIntyre, I., Zaidi, M., Towhidul Alam, A. S. M., Datta, H. K., Moonga, B. S., Lidbury, P. S., Hecker, M., and Vane, J. R., Osteoclastic inhibition: An action of nitric oxide not mediated by cyclic GMP. Proc. Natl. Acad. Sci. USA, 88, 2936-2940 (1991) https://doi.org/10.1073/pnas.88.7.2936
  13. Messina, M. and Messina, V., Soyfood, soybean isoflavone, and bone health: a brief overview. J. Ren. Nutri., 10, 63-68 (2000) https://doi.org/10.1016/S1051-2276(00)90001-3
  14. Nakagawa, N., Kinosaki, M., Yamaguchi, K., Shima, N., Yasuda, H., Yano, K., Morinaga, T., and Higashio, K., RANK is the essential signaling receptor for osteoclast differentiation factor in osteoclastogenesis. Biochem. Biophys. Res. Commun., 253, 395-400 (1998) https://doi.org/10.1006/bbrc.1998.9788
  15. Pacifici, R., Cytokines, estrogen and postmenopausal osteoporosis- the second decade. Endocrinology, 139, 2659-2661 (1998) https://doi.org/10.1210/en.139.6.2659
  16. Ragab, A. A., Nalepka, J. L., Bi, Y., and Greenfield, E. M., Cytokines synergistically induce osteoclast differentiation: support by immortalized or normal calvarial cells. Am. J. Physiol. Cell Physiol., 283, C679-C687 (2002) https://doi.org/10.1152/ajpcell.00421.2001
  17. Raisz, L. G., The osteoporosis revolution. Ann. Intern. Med., 126, 458-462 (1997) https://doi.org/10.7326/0003-4819-126-6-199703150-00007
  18. Riggs, B. L., Khosia, S., and Melton, L. J., A unitary model for involutional osteoporosis: estrogen deficiency causes both type I and type II osteoporosis in postmenopausal women and contributes to bone loss in aging men. J. Bone Miner. Res., 13, 763-773 (1998) https://doi.org/10.1359/jbmr.1998.13.5.763
  19. Setchell, K. D., Abstract presented at: Second international symposium on the role of soy in preventing and treating chronic disease. Brussel Belgium , (1996)
  20. Stanley, E. R. and Heard, P. M., Factors regulating macrophage production and growth: Purification and some properties of the colony-stimulating factor from medium conditioned by mouse L cells. J. Biol. Chem., 252, 4305-4321 (1977)
  21. Swolin-Eide, D. and Ohlsson, C., Effects of cortisol on the expression of interleukin-6 and interleukin-1 in human osteoblast like cells. J. Endocrinology, 156, 107-114 (1998) https://doi.org/10.1677/joe.0.1560107
  22. Tai, H., Miyaura, C., Pilbeam, C. C., Tamura, T., Ohsugi, Y., Koishihara, Y., Kubodera, N., Kawaguchi, H., Raisz, L. G., and Suda, T., Transcriptional induction of cyclooxygenase-2 in osteoblast is involved in interleukin-6-induced osteocalst formation. Endocrinology, 138, 2372-2397 (1997) https://doi.org/10.1210/en.138.6.2372
  23. Tamura, T., Udagawa, N., Takahashi, N., Miyaura, C., Tanaka, S., Yamada, Y., Akatsu, T., Koishihara, Y., Ohsugi, Y., Kumaki, K., Taga, T., Kishmoto, T., and Suda, T., Soluble interleukin-6 receptor triggers osteoclast formation by interleukin-6. Proc. Natl. Acad. Sci. USA, 90, 11924-11928 (1993) https://doi.org/10.1073/pnas.90.24.11924
  24. Udagawa, N., Takahashi, N., Matsuzaki, K., Jimi, E., Tsurukai, T., Itoh, K., Nakagawa, H., Yasuda, H., Goto, M., Tsuda, E., Higashio, K., Martin, T. J., and Suda, T., Osteoblasts/stromal cells stimulate osteoclast activation through expression of osteoclast differentiation factor/RANKL but not macrophage colony-stimulating factor: receptor activator of NF-kappa B ligand. Bone, 5, 517-523 (1999)
  25. vant Hof, R. J. and Ralston, S. H., Nitric oxide and bone. Immunology, 103, 255-261 (2001) https://doi.org/10.1046/j.1365-2567.2001.01261.x
  26. Wimalawansa, S. J., De Marco, G., Gangula, P., and Yallampalli, Nitric oxide donor alleviates ovariectomy-induced bone loss. Bone, 18, 301-304 (1996) https://doi.org/10.1016/8756-3282(96)00005-1
  27. Yamaguchi, M. and Gao, Y. H., Inhibitory effect of genistein on bone resorption in tissue culture. Biochem. Phamacol., 55, 71-76 (1998) https://doi.org/10.1016/S0006-2952(97)00402-4