KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지

Paraquat Induced Heme Oxygenase-1 in Dopaminergic Cells

도파민 세포에서 Paraquat에 의한 헴산화효소-1의 유도

  • Chun Hong Sung (Department of Biotechnology, College of Natural Sciences and Research Center for Proteineous Materials, Chosun University)
  • 전홍성 (조선대학교 자연과학대학 생명공학과, 단백질소재연구센터)
  • Published : 2005.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Paraquat, a widely used herbicide, has been suggested as a potential risk factor for Parkinson's disease. Heme oxygenase-1(HO-1), a marker for oxidative stress and endoplasmic reticulum(ER) stress, is known to catalyze heme to biliverdin, carbon monoxide and free iron in response to various stimuli. Here we show that paraquat activates HO-1 expression in a time-and dose-dependent manner in substantia nigra(SN) dopaminergic neuronal cells. Activation of Ho-1 by paraquat was regulated primarily at the level of gene transcription. Deletion analysis of the promoter and the 5' distal enhancers, E1 and E2, of the HO-1 gene revealed that the E2 enhancer is a potent inducer of the paraquat-dependent Ho-1 gene expression in dopamninergic neuronal cells. Mutational analysis of the E2 enhacer further demonstrated that the transcription factor activator protein-1(AP-1) plays an important role in mediating paraquat-induced HO-1 gene transcription. Moreover, using specific inhibitors of the mitogen-activated protein kinases(MAPKs), we investigated the role of paraquat and MAPKs for HO-1 gene regulation in dopaminergic cells. The c-Jun N-terminal kinase(JNK) inhibitor SP600125 significantly suppressed the expression of HO-1 by paraquat. All these results demonstrate that induction of HO-1 by paraquat requies the activation of the AP-1 and JNK pathway.

흔하게 사용되어온 제초제인 paraquat는 파킨슨병의 원인이 될 수 있는 유력한 위험 요소이다. 헴산화효소-1(HO-1)은 산화적 스트레스와 소포체 스트레스의 marker인데, 여러 가지 자극에 의해 heme을 분해하여 biliverdin, 일산화탄소, 철 성분으로 전환시킨다 본 연구에서는 뇌의 흑색질 유래의 도파민 세포주 SN4741에서 paraquat가 시간별, 농도별로 HO-1을 활성화시키는 기작을 조사하였다. HO-1이 Paraquat에 의해 활성화되는 것은 주로 유전자 전사 수준에서 조절되었다. HO-1 유전자의 promoter와 5' enhancer인 El, E2를 결실시킨 실험에서, E2 enhancer가 도파민 세포에서 paraquat에 의한 HO-1 유전자 발현을 유도하는 핵심 부위로 판명되었다 E2 enhancer 부위를 돌연변이 시킨 실험 결과는 전사인자 활성 단백질-1 (AP-1) 결합부위를 통해 HO-1 발현이 유도됨을 밝히게 되었다. 또한, 도파민 세포에서 HO-1 유전자 발현의 조절과 신호전달 과정의 관계를 조사하기 위해 MAP kinase들의 특이적 저해제를 처리하고 paraquat로 자극을 준 결과, JNK 저해제인 SP600125가 가장 현저하게 paraquat에 의한 HO-1 발현을 억제하였다. 결론적으로, 도파민 세포에서 paraquat가 HO-1을 유도하는 데는 E2 enhancer가 중요하게 작용하고, AP-1과 JNK 경로를 통해 HO-1 발현이 조절된다는 사실을 처음으로 밝히게 되었다.

Keywords

References

  1. Tanner, C. M. and Y. Ben-Shlomo (1999), Epidemiology of Parkinson's disease, Adv. Neurol. 80, 153-159
  2. McCormack, A L., M. Thiruchelvam, A. B. Manning-Bog, C. Thiffault, J. W. Langston, D. A. Cory-Slechta, and D. A. Di Monte (2002), Environmental risk factors and Parkinson's disease: selective degeneration of nigral dopaminergic neurons caused by the herbicide paraquat, Neurobiol. Dis. 10, 119-127 https://doi.org/10.1006/nbdi.2002.0507
  3. Liou, H. H., M. C. Tsai, C. J. Chen, J. S. Jeng, Y. C. Chang, S. Y. Chen, and R. C. Chen (1997), Environmental risk factors and Parkinson's disease: a case-control study in Taiwan, Neurology. 48, 1583-1588 https://doi.org/10.1212/WNL.48.6.1583
  4. Mollace, V., M. Iannone, C. Muscoli, E. Palma, T. Granato, V. Rispoli, R. Nistieo, D. Rotiroti, and D. Salvemini (2003), The role of oxidative stress in paraquat-induced neurotoxicity in rats: protection by non-peptidyl superoxide dismutase mimetic, Neurosci. Lett. 335, 163-166 https://doi.org/10.1016/S0304-3940(02)01168-0
  5. Shimizu, K., K. Matsubara, K. Ohtaki, S. Fujumaru, O. Saito, and H. Shiono (2003), Paraquat induces long-lasting dopamine overflow through the excitotoxic pathway in the striatum of freely moving rats, Brain Res. 976, 243-252 https://doi.org/10.1016/S0006-8993(03)02750-1
  6. Manning-Bog, A. B., A. L. McConnack, J. Li, V. N. Uversky, A. L. Fink, and D. A. Di Monte (2002), The herbicide paraquat causes up-regulation and aggregation of $\alpha$-synuclein in mice: paraquat and $\alpha$-synuclein, J. BioI. Chem. 277, 1641-1644 https://doi.org/10.1074/jbc.C100560200
  7. Schipper, H. M., A. Liberman, and E. G. Stopa (1998), Neural heme oxygenase-l expression in idiopathic Parkinson's disease, Exp. Neurol. 150, 60-68 https://doi.org/10.1006/exnr.1997.6752
  8. Linden, T., J. Doutheil, and W. Paschen (1998), Role of calcium in the activation of erp72 and heme oxygenase-l expression on depletion of endoplasmic reticulum calcium stores in rat neuronal cell culture, Neurosci. Lett. 247, 103-106 https://doi.org/10.1016/S0304-3940(98)00278-X
  9. Hartsfield, C. L., J. Alam, and A. M. Choi (1998), Transcriptional regulation of the heme oxygenase 1 gene by pyrrolidine dithiocarbamate, FASEB J. 12, 1675-1682 https://doi.org/10.1096/fasebj.12.15.1675
  10. Tenhunen R., H. S. Marver, and R. Schmid (1969), Microsomal heme oxygenase: characterization of theenzyme, J. Biol. Chem. 244, 6388-6394
  11. Ewing, J. F. and M. D. Maines(1991), Rapidinductionof heme oxygenase 1 mRNAand proteinby hyperthermia in rat brain:hemeoxygenase 2 is not a heat shock protein, Proc. Natl. Acad Sci. USA. 88, 5364-5368
  12. Alam, J., S. Camhi, and A. M. Choi (1995), Identification of a second region upstreamof the mouse heme oxygenase-l gene that functions as a basal level and inducer-dependent transcription enhancer, J. Biol. Chem. 270, 11977-11984 https://doi.org/10.1074/jbc.270.20.11977
  13. Camhi, S. L., J. Alam, G. W. Wiegand, B. Y. Chin, and A. M. Choi (1998), Transcriptional activation of the HO-1 gene by lipopolysaccharide is mediatedby 5' distal enhancers: role of reactiveoxygenintermediates and AP-1, Am. J. Respir. Cell Mol. BioI. 18,226-234 https://doi.org/10.1165/ajrcmb.18.2.2910
  14. Chun, H. S., G. E. Gibson, L. A. DeGiorgio, H. Zhang,V. J. kidd,and J. H. Son (2001), Dopaminergic cell death induced by MPP+, oxidant and specific neurotoxicants shares the common molecular mechanism, J. Neurochem. 76,1010-1021 https://doi.org/10.1046/j.1471-4159.2001.00096.x
  15. Motterlini, R., R. Foresti, M. Intaglietta, and R. M. Winslow (1996), NO-mediated activation of heme oxygenase: endogenous cytoprotection against oxidative stress to endothelium, Am. J. Physiol. Heart Circ. Physiol. 270, 107-114 https://doi.org/10.1152/ajpcell.1996.270.1.C107
  16. Alam, J., C. Wicks, D. Stewart, P. Gong, C. Touchard, S. Otterbein, A. M. Choi, M. E. Burow,and J. Tou (2000), Mechanism of heme oxygenase-I gene activation by cadmiumin MCF-7mammary epithelial cells. Role of p38 kinaseand Nrf2 transcription factor, J. Biol. Chem. 275,27694-27702
  17. Maines, M. D. (1997), The heme oxygenase system: a regulatorof second messenger gases, Annu. Rev. Pharmacol. Toxicol. 37,517-554 https://doi.org/10.1146/annurev.pharmtox.37.1.517
  18. Choi, A. M. and J. Alam (1996), Heme oxygenase-I: function, regulation, and implication of a novelstress-inducible proteinin oxidant-induced lung injury, Am. J. Respir. Cell Mol. BioI. 15,9-19 https://doi.org/10.1165/ajrcmb.15.1.8679227
  19. Kiemer, A. K., N. Bildner, N. C. Weber, and A. M. Vollmar (2003), Characterization of hemeoxygenase1 (heat shock protein 32) induction by atrial natriuretic peptide in human endothelial cells, Endocrinology. 144, 802-812 https://doi.org/10.1210/en.2002-220610
  20. Hunter, T. and M. Karin (1992), The regulation of transcription by phosphorylation, Cell. 70, 375-387 https://doi.org/10.1016/0092-8674(92)90162-6
  21. Kietzmann, T., A. Samoylenko, and S. Immenschuh (2003), Transcriptional regulation of heme oxygenase-l gene expression by MAP kinases of the JNK and p38 pathways in primary cultures of rat hepatocytes, J. BioI. Chem. 278, 17927-17936 https://doi.org/10.1074/jbc.M203929200
  22. Brooks, A. I., C. A. Chadwick, H. A. Gelbard, D. A. Cory-Slechta, and H. J. Federoff (1999), Paraquat elicited neurobehavioral syndrome caused by dopaminergic neuron loss, Brain Res. 823, 1-10 https://doi.org/10.1016/S0006-8993(98)01192-5