DOI QR코드

DOI QR Code

Involvement of Oxidative Stress in Formaldehyde-induced Apoptosis in Cultured Lung Macrophage Cells

폐 대식세포주에서 포름알데히드에 의한 세포 사멸 효과에 대한 산화성 스트레스 관련성

  • Park, Soo-Hyun (Bio-therapy Human Resources Center, Animal Medical Center, Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University)
  • 박수현 (전남대학교 수의과대학 바이오치료 산업인력 양성팀, 동물의학 연구소)
  • Published : 2009.09.30

Abstract

Formaldehyde (FA) is an important irritant compound in pesticide to induce asthma and allergy in respiratory system. Alveolar macrophage is also an pivotal cell in the immune response of respiratory system. However, the effect of FA in macrophage cell viability has not been elucidated. Thus, this study was conducted to investigate the effect of FA on apoptosis in Raw 264.7 cells, alveolar macrophage cell line. In this study, FA decreased cell viability of lung alveolar macrophage cells in a dose-dependent manner (>$100{\mu}M$). FA-induced decrease of cell viability was blocked by the treatment of antioxidants (vitamin C, NAC, and catalase). Indeed, FA induced lipid peroxide formation in Raw 264.7 cells. FA decreased Bcl-2 expression but increased Bax expression in lung alveloar macrophage cells. In addition, FA also increased the cleaved form of caspase-3. In conclusion, FA induced apoptosis via oxidative stress in cultured Raw 264.7 cells.

포름알데히드는 농약 노출 시에 나타나는 중요한 물질로 천식 및 알러지등의 호흡 질환을 일으키는 물질로 알려져 있으며, 폐에서 대식세포는 면역 반응에 있어서 방어 기능을 담당하는 세포로 알려져 있다. 그러나 대식세포에서 포름알데히드에 대한 효과는 알려져 있지 않고 있어서 대식 세포주인 Raw 264.7 세포를 이용하여 실험하였다. 실험 결과 포름알데히드는 세포 생존율을 감소 시켰으며, 이러한 반응은 항산화제인 vitamin C, NAC, 및 catalase 처리 시 차단되었다. 실제로 포름알데히드 처리시 산화성 스트레스 지표인 lipid peroxide 형성이 증가하였으며 이들 반응 역시 항산화제들에 의해 차단되었다. 한편 포름알데히드 처리시 세포 사멸 촉진 단백질인 Bax 발현은 증가하였으며 세포 사멸 억제 단백질인 Bcl-2의 발현은 억제되었으며 이러한 반응은 항산화제 처리시 차단되었다. 세포사멸 실행 단백질인 casapse-3의 활성형 역시 증가하였으며, 항산화제 처리시 차단되었다. 결론적으로 포름알데히드는 폐 대식세포에서 산화성 스트레스 증가를 통해 세포 사멸을 일으키는 것으로 나타났다.

Keywords

References

  1. Kampa, M. and Castanas, E. (2008) Human health effects of air pollution. Environ Pollut. 151(2), 362-367 https://doi.org/10.1016/j.envpol.2007.06.012
  2. Kita, T., Fujimura, M., Myou, S., Ishiura, Y., Abo, M., Katayama, N., Nishitsuji, M., Yoshimi, Y., Nomura, S., Oribe, Y., and Nakao, S. (2003) Potentiation Potentiation of allergic bronchoconstriction by repeated exposure to formaldehyde in guinea-pigs in vivo. Clin Exp Allergy 33, 1747-1753 https://doi.org/10.1111/j.1365-2222.2003.01826.x
  3. Nakazawa, H., Ikeda, H., Yamashita, T., Hara, I., Kumai, Y., Endo, G. and Endo, Y. (2005) A case of sick building syndrome in a Japanese office worker. Ind Health. 43, 341-345 https://doi.org/10.2486/indhealth.43.341
  4. Bagchi, D., Bagchi, M., Hassoun, E.A., and Stohs, S. J. (1993) Detection of paraquat-induced in vivo lipid peroxidation by gas chromatography/mass spectrometry and high-pressure liquid chromatography. J. Anal. Toxicol. 17(7), 411-414 https://doi.org/10.1093/jat/17.7.411
  5. Callas, P.W., Pastides, H., and Hosmer, D. W. Jr. (1996) Lung cancer mortality among workers in formaldehyde industries. J. Occup. Environ. Med. 38(8), 747-748 https://doi.org/10.1097/00043764-199608000-00006
  6. Collins, J.J., Acquavella, J.F., and Esmen, N.A. (1997) An updated meta-analysis of formaldehyde exposure and upper respiratory tract cancers. J. Occup. Environ. Med. 39(7), 639-651 https://doi.org/10.1097/00043764-199707000-00009
  7. Marsh, G.M., Stone, R.A., Esmen, N.A., Henderson, V.L., and Lee, K.Y. (1996) Mortality among chemical workers in a factory where formaldehyde was used. Occup. Environ. Med. 53(9), 613-627 https://doi.org/10.1136/oem.53.9.613
  8. Kamdar, O., Le, W., Zhang, J., Ghio, A.J., Rosen, G. D, and Upadhyay, D. (2008) Air pollution induces enhanced mitochondrial oxidative stress in cystic fibrosis airway epithelium. FEBS Lett. 582, 3601-3606 https://doi.org/10.1016/j.febslet.2008.09.030
  9. Schneider, J. C., Card, G. L., Pfau, J. C., and Holian, A. (2005) Air pollution particulate SRM 1648 causes oxidative stress in RAW 264.7 macrophages leading to production of prostaglandin E2, a potential Th2 mediator. Inhal. Toxicol. 17, 871-877 https://doi.org/10.1080/08958370500244498
  10. Higychi, M., Hisgahi, N., Taki, H., and Osawa, T. (1990) Cytolytic mechanisms of activ ated macrophages. Tumor necrosis facto and L-arginine-dependent mechani는 act synergistically as the major cytolytic mechani는 of activated macrophages. J. Immunology. 144, 1425-1431
  11. Khadaroo, R. G., Kapus, A., Powers, K.A., Cybulsky, M. I., Marshall, J. C., and Rotstein, O.D. (2003) Oxidative stress reprograms lipopolysaccharide signaling via Src kinase-dependent pathway in RAW 264.7 macrophage cell line. J. Biol. Chem. 278, 47834-47841 https://doi.org/10.1074/jbc.M302660200
  12. Imrich, A., Ning, Y., Lawrence, J., Coull, B., Gitin, E., Knutson, M., and Kobzik L. (2007) Alveolar macrophage cytokine response to air pollution particles: oxidant mechanisms. Toxicol. Appl. Pharmacol. 218, 256-264 https://doi.org/10.1016/j.taap.2006.11.033
  13. Zinkel, S., Gross, A. and Yang, E. (2006) BCL2 family in DNA damage and cell cycle control. Cell Death Differ. 13(8), 1351-1359 https://doi.org/10.1038/sj.cdd.4401987
  14. Kutuk, O. and Basaga, H. (2007) Apoptosis signalling by 4-hydroxynonenal: a role for JNK-c-Jun/AP-1 pathway. Redox Rep. 12(1), 30-34 https://doi.org/10.1179/135100007X162329
  15. Zakeri, Z. and Lockshin, R. A. (2008) Cell death: history and future. Adv Exp Med Biol. 615, 1-11 https://doi.org/10.1007/978-1-4020-6554-5_1
  16. Fujimaki, H., Kurokawa, Y., Kunugita, N., Kikuchi, M., Sato, F. and Arashidani, K. (2004) Differential immunogenic and neurogenic inflammatory responses in an allergic mouse model exposed to low levels of formaldehyde. Toxicology. 197, 1-13 https://doi.org/10.1016/j.tox.2003.11.015
  17. Nakao, H., Umebayashi, C., Nakata, M., Nishizaki, Y., Noda, K., Okano, Y., and Oyama, Y. (2003) Formaldehyde-induced shrinkage of rat thymocytes. J. Pharmacol. Sci. 91(1), 83-86 https://doi.org/10.1254/jphs.91.83
  18. Grafstrom, R. C., Fornace, A. J. Jr, Autrup, H., Lechner, J. F., and Harris, C. C. (1983) Formaldehyde damage to DNA and inhibition of DNA repair in human bronchial cells. Science. 220, 216-218 https://doi.org/10.1126/science.6828890
  19. Shang, Y., Li, X., Prasad, P. V., Xu, S., Yao, S., Liu, D., Yuan, S., and Feng, D. (2009) Erythropoietin attenuates lung injury in lipopolysaccharide treated rats. J. Surg. Res. 155, 104-110 https://doi.org/10.1016/j.jss.2008.10.003
  20. Wu, Y., Xing, D., Chen, W.R. and Wang, X. (2007) Bid is not required for Bax translocation during UV-induced apoptosis. Cell Signal. 19(12), 2468-2478 https://doi.org/10.1016/j.cellsig.2007.07.024
  21. Cao, X., Bennett, R. L. and May, W. S. (2008) c-Myc and caspase-2 are involved in activating Bax during cytotoxic drug-induced apoptosis. J. Biol. Chem. 283(21), 14490-14496 https://doi.org/10.1074/jbc.M801107200
  22. Tong, Q. S., Zheng, L. D., Tang, S. T., Jiang, G. S., Ruan, Q. L., Zeng, F. Q., and Dong, J. H. (2007) Nitrofen suppresses cell proliferation and promotes mitochondria-mediated apoptosis in type II pneumocytes. Acta Pharmacol Sin. 28(5), 672-684 https://doi.org/10.1111/j.1745-7254.2007.00552.x
  23. An, S., Hishikawa, Y., Liu, J., and Koji, T. (2007) Lung injury after ischemia-reperfusion of small intestine in rats involves apoptosis of type II alveolar epithelial cells mediated by TNF-alpha and activation of Bid pathway. Apoptosis. 12(11), 1989-2001 https://doi.org/10.1007/s10495-007-0125-1
  24. Tsukahara, S., Yamamoto, S., Tin-Tin-Win-Shwe, Ahmed, S., Kunugita, N., Arashidani, K., and Fujimaki, H. (2006) Inhalation of low-level formaldehyde increases the Bcl-2/Bax expression ratio in the hippocampus of immunologically sensitized mice. Neuroimmunomodulation. 13(2), 63-68 https://doi.org/10.1159/000094829
  25. Susnow, N., Zeng, L., Margineantu, D., and Hockenbery DM. (2009) Bcl-2 family proteins as regulators of oxidative stress. Semin. Cancer Biol. 19(1), 42-49 https://doi.org/10.1016/j.semcancer.2008.12.002