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Arecoline Induces Neurotoxicity to PC12 Cells: Involvement in ER Stress and Disturbance of Endogenous H2S Generation

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Abstract

Arecoline is a major alkaloid of areca nut and has been effect on central nervous system. Although arecoline-induced neurotoxicity has been reported, the possible underlying neurotoxic mechanisms have not yet been elucidated. Increasing evidences have shown that both excessive endoplasmic reticulum (ER) stress and disturbance of hydrogen sulfide (H2S) production are involved in the pathophysiology of numerous neurodegenerative diseases. Here, the purpose of present study was to verify whether ER stress and the disturbance of endogenous H2S generation are also involved in arecoline-caused neurotoxicity. We found that treatment of PC12 cells with arecoline induced the down-regulation of cells viability and up-regulation of apoptosis and the activity of caspase-3, indicating the neurotoxic role of arecoline to PC12 cells. In addition, arecoline also increased the expression of Bax (pro-apoptotic protein) and attenuated the expression of Bcl-2 (anti-apoptotic protein) in PC12 cells. Simultaneously, arecoline caused excessive ER stress in PC12 cells, as evidenced by the up-regulations of Glucose-regulated protein 78 (GRP78), CCAAT/enhancer binding protein homologous protein (CHOP), and Cleaved caspase-12 expressions. Notably, the level of H2S in the culture supernatant and the expressions of cystathionine β-synthase and 3-mercaptopyruvate sulfurtransferase (two major enzymes for endogenous H2S generation in PC12 cells) were also reduced by arecoline treatment. These results indicate that arecoline-caused neurotoxicity to PC12 cells is involved in ER stress and disturbance of endogenous H2S generation and suggest that the modulation of ER stress and endogenous H2S generation may be potential therapeutic approach in treatment of arecoline-caused neurotoxicity.

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References

  1. Garg A, Chaturvedi P, Gupta PC (2014) A review of the systemic adverse effects of areca nut or betel nut. Indian J Med Paediatr Oncol: Off J Indian Soc Med Paediatr Oncol 35:3–9

    Article  Google Scholar 

  2. Dasgupta R, Saha I, Pal S, Bhattacharyya A, Sa G, Nag TC, Das T, Maiti BR (2006) Immunosuppression, hepatotoxicity and depression of antioxidant status by arecoline in albino mice. Toxicology 227:94–104

    Article  CAS  PubMed  Google Scholar 

  3. Khan S, Chatra L, Prashanth SK, Veena KM, Rao PK (2012) Pathogenesis of oral submucous fibrosis. J Cancer Res Ther 8:199–203

    Article  PubMed  Google Scholar 

  4. Choudhury MD, Chetia P, Choudhury KD, Talukdar AD, Datta-Choudhari M (2012) Atherogenic effect of arecoline: a computational study. Bioinformation 8:229–232

    Article  PubMed  PubMed Central  Google Scholar 

  5. Thangjam GS, Kondaiah P (2009) Regulation of oxidative-stress responsive genes by arecoline in human keratinocytes. J Periodontal Res 44:673–682

    Article  CAS  PubMed  Google Scholar 

  6. Hung KF, Lai KC, Liu TY, Liu CJ, Lee TC, Lo JF (2009) Asb6 upregulation by Areca nut extracts is associated with betel quid-induced oral carcinogenesis. Oral Oncol 45:543–548

    Article  CAS  PubMed  Google Scholar 

  7. Sadashiva CT, Chandra JN, Kavitha CV, Thimmegowda A, Subhash MN, Rangappa KS (2009) Synthesis and pharmacological evaluation of novel N-alkyl/aryl substituted thiazolidinone arecoline analogues as muscarinic receptor 1 agonist in Alzheimer’s dementia models. Eur J Med Chem 44:4848–4854

    Article  CAS  PubMed  Google Scholar 

  8. Bales A, Peterson MJ, Ojha S, Upadhaya K, Adhikari B, Barrett B (2009) Associations between betel nut (Areca catechu) and symptoms of schizophrenia among patients in Nepal: a longitudinal study. Psychiatry Res 169:203–211

    Article  PubMed  Google Scholar 

  9. Monmaur P, Allix M, Schoevaert-Brossault D, Houcine O, Plotkine M, Willig F (1990) Effects of transient cerebral ischemia on the hippocampal dentate theta (theta) profile in the acute rat: a study 4–5 months following recirculation. Brain Res 508:124–134

    Article  CAS  PubMed  Google Scholar 

  10. Shih YT, Chen PS, Wu CH, Tseng YT, Wu YC, Lo YC (2010) Arecoline, a major alkaloid of the areca nut, causes neurotoxicity through enhancement of oxidative stress and suppression of the antioxidant protective system. Free Radic Biol Med 49:1471–1479

    Article  CAS  PubMed  Google Scholar 

  11. Gething MJ, Sambrook J (1992) Protein folding in the cell. Nature 355:33–45

    Article  CAS  PubMed  Google Scholar 

  12. Ma Y, Hendershot LM (2004) ER chaperone functions during normal and stress conditions. J Chem Neuroanat 28:51–65

    Article  CAS  PubMed  Google Scholar 

  13. Placido AI, Pereira CM, Duarte AI, Candeias E, Correia SC, Carvalho C, Cardoso S, Oliveira CR, Moreira PI (2015) Modulation of endoplasmic reticulum stress: an opportunity to prevent neurodegeneration? CNS Neurol Disord: Drug Targets 14:518–533

    Article  CAS  Google Scholar 

  14. Torres M, Matamala JM, Duran-Aniotz C, Cornejo VH, Foley A, Hetz C (2015) ER stress signaling and neurodegeneration: at the intersection between Alzheimer’s disease and prion-related disorders. Virus Res 207:69–75

    Article  CAS  PubMed  Google Scholar 

  15. Chaudhari N, Talwar P, Parimisetty A, Lefebvre d’Hellencourt C, Ravanan P (2014) A molecular web: endoplasmic reticulum stress, inflammation, and oxidative stress. Front Cell Neurosci 8:213

    Article  PubMed  PubMed Central  Google Scholar 

  16. Fung TS, Liu DX (2014) Coronavirus infection, ER stress, apoptosis and innate immunity. Front Microbiol 5:296

    Article  PubMed  PubMed Central  Google Scholar 

  17. Jin ML, Park SY, Kim YH, Oh JI, Lee SJ, Park G (2014) The neuroprotective effects of cordycepin inhibit glutamate-induced oxidative and ER stress-associated apoptosis in hippocampal HT22 cells. Neurotoxicology 41:102–111

    Article  CAS  PubMed  Google Scholar 

  18. Goswami P, Gupta S, Biswas J, Sharma S, Singh S (2015) Endoplasmic reticulum stress instigates the rotenone induced oxidative apoptotic neuronal death: a study in rat brain. Mol Neurobiol. doi:10.1007/s12035-015-9463-0

  19. Wu CX, Liu R, Gao M, Zhao G, Wu S, Wu CF, Du GH (2013) Pinocembrin protects brain against ischemia/reperfusion injury by attenuating endoplasmic reticulum stress induced apoptosis. Neurosci Lett 546:57–62

    Article  CAS  PubMed  Google Scholar 

  20. Kim I, Xu W, Reed JC (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nat Rev Drug Discov 7:1013–1030

    Article  CAS  PubMed  Google Scholar 

  21. Zhang X, Bian JS (2014) Hydrogen sulfide: a neuromodulator and neuroprotectant in the central nervous system. ACS Chem Neurosci 5:876–883

    Article  CAS  PubMed  Google Scholar 

  22. Abe K, Kimura H (1996) The possible role of hydrogen sulfide as an endogenous neuromodulator. J Neurosci 16:1066–1071

    CAS  PubMed  Google Scholar 

  23. Zhang Q, Yuan L, Liu D, Wang J, Wang S, Zhang Q, Gong Y, Liu H, Hao A, Wang Z (2014) Hydrogen sulfide attenuates hypoxia-induced neurotoxicity through inhibiting microglial activation. Pharmacol Res 84:32–44

    Article  CAS  PubMed  Google Scholar 

  24. Li MH, Tang JP, Zhang P, Li X, Wang CY, Wei HJ, Yang XF, Zou W, Tang XQ (2014) Disturbance of endogenous hydrogen sulfide generation and endoplasmic reticulum stress in hippocampus are involved in homocysteine-induced defect in learning and memory of rats. Behav Brain Res 262:35–41

    Article  CAS  PubMed  Google Scholar 

  25. Nie L, Hu Y, Yan X, Li M, Chen L, Li H, Li X, Zhou H, Zheng Y (2013) The anti-apoptotic effect of hydrogen sulfide attenuates injuries to the medullary respiratory centers of neonatal rats subjected to in utero cigarette smoke exposure. Respir Physiol Neurobiol 188:29–38

    Article  CAS  PubMed  Google Scholar 

  26. Dwyer BE, Raina AK, Perry G, Smith MA (2004) Homocysteine and Alzheimer’s disease: a modifiable risk? Free Radic Biol Med 36:1471–1475

    Article  CAS  PubMed  Google Scholar 

  27. Du C, Jin M, Hong Y, Li Q, Wang XH, Xu JM, Wang F, Zhang Y, Jia J, Liu CF, Hu LF (2014) Downregulation of cystathionine beta-synthase/hydrogen sulfide contributes to rotenone-induced microglia polarization toward M1 type. Biochem Biophys Res Commun 451:239–245

    Article  CAS  PubMed  Google Scholar 

  28. Tang XQ, Fang HR, Zhou CF, Zhuang YY, Zhang P, Gu HF, Hu B (2013) A novel mechanism of formaldehyde neurotoxicity: inhibition of hydrogen sulfide generation by promoting overproduction of nitric oxide. PLoS ONE 8:e54829

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Doeller JE, Isbell TS, Benavides G, Koenitzer J, Patel H, Patel RP, Lancaster JR Jr, Darley-Usmar VM, Kraus DW (2005) Polarographic measurement of hydrogen sulfide production and consumption by mammalian tissues. Anal Biochem 341:40–51

    Article  CAS  PubMed  Google Scholar 

  30. Cohen GM (1997) Caspases: the executioners of apoptosis. Biochem J 326(Pt 1):1–16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Logue SE, Cleary P, Saveljeva S, Samali A (2013) New directions in ER stress-induced cell death. Apoptosis 18:537–546

    Article  PubMed  Google Scholar 

  32. Hetz C (2012) The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol 13:89–102

    CAS  PubMed  Google Scholar 

  33. Zhou CF, Tang XQ (2011) Hydrogen sulfide and nervous system regulation. Chin Med J 124:3576–3582

    CAS  PubMed  Google Scholar 

  34. Kimura H (2013) Physiological role of hydrogen sulfide and polysulfide in the central nervous system. Neurochem Int 63:492–497

    Article  CAS  PubMed  Google Scholar 

  35. Winstock A (2002) Areca nut-abuse liability, dependence and public health. Addict Biol 7:133–138

    Article  CAS  PubMed  Google Scholar 

  36. Li M, Gao F, Zhou ZS, Zhang HM, Zhang R, Wu YF, Bai MH, Li JJ, Lin SR, Peng JY (2014) Arecoline inhibits epithelial cell viability by upregulating the apoptosis pathway: implication for oral submucous fibrosis. Oncol Rep 31:2422–2428

    CAS  PubMed  Google Scholar 

  37. Radi E, Formichi P, Battisti C, Federico A (2014) Apoptosis and oxidative stress in neurodegenerative diseases. J Alzheimer’s Dis 42(Suppl 3):S125–S152

    Google Scholar 

  38. Fadeel B, Ottosson A, Pervaiz S (2008) Big wheel keeps on turning: apoptosome regulation and its role in chemoresistance. Cell Death Differ 15:443–452

    Article  CAS  PubMed  Google Scholar 

  39. Cheng HL, Su SJ, Huang LW, Hsieh BS, Hu YC, Hung TC, Chang KL (2010) Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis-involvement of STAT3 and RhoA activation. Mol Cancer 9:126

    Article  PubMed  PubMed Central  Google Scholar 

  40. Stefani IC, Wright D, Polizzi KM, Kontoravdi C (2012) The role of ER stress-induced apoptosis in neurodegeneration. Curr Alzheimer Res 9:373–387

    Article  CAS  PubMed  Google Scholar 

  41. Luo Y, Yang X, Zhao S, Wei C, Yin Y, Liu T, Jiang S, Xie J, Wan X, Mao M, Wu J (2013) Hydrogen sulfide prevents OGD/R-induced apoptosis via improving mitochondrial dysfunction and suppressing an ROS-mediated caspase-3 pathway in cortical neurons. Neurochem Int 63:826–831

    Article  CAS  PubMed  Google Scholar 

  42. Yin J, Tu C, Zhao J, Ou D, Chen G, Liu Y, Xiao X (2013) Exogenous hydrogen sulfide protects against global cerebral ischemia/reperfusion injury via its anti-oxidative, anti-inflammatory and anti-apoptotic effects in rats. Brain Res 1491:188–196

    Article  CAS  PubMed  Google Scholar 

  43. Eto K, Asada T, Arima K, Makifuchi T, Kimura H (2002) Brain hydrogen sulfide is severely decreased in Alzheimer’s disease. Biochem Biophys Res Commun 293:1485–1488

    Article  CAS  PubMed  Google Scholar 

  44. Tan H, Zou W, Jiang J, Tian Y, Xiao Z, Bi L, Zeng H, Tang X (2015) Disturbance of hippocampal H2S generation contributes to CUMS-induced depression-like behavior: involvement in endoplasmic reticulum stress of hippocampus. Acta Biochim et Biophys Sin 47:285–291

    Article  CAS  Google Scholar 

  45. Lin F, Chen Y, Liao C, Sun Y, Bai Y, Liao Y, Li M, Qi Y (2015) Hydrogen sulfide attenuates bronchial epithelial cell apoptosis by inhibiting endoplasmic reticulum stress. Zhonghua yi xue za zhi 95:2297–2301

    CAS  PubMed  Google Scholar 

  46. Ying R, Wang XQ, Yang Y, Gu ZJ, Mai JT, Qiu Q, Chen YX, Wang JF (2016) Hydrogen sulfide suppresses endoplasmic reticulum stress-induced endothelial-to-mesenchymal transition through Src pathway. Life Sci 144:208–217

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by Natural Science Foundation of China (81202518), Zheng xiang Scholar Program of University of South China (2014-004), the construct program of the key discipline in Hunan province.

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    Authors and Affiliations

    1. Department of Cardiology, The Affiliated Hospital, Guangdong Medical University, 57 W Friendship Street People Avenue South, Zhanjiang, 524001, Guangdong, People’s Republic of China

      Jia-Mei Jiang, Keng Wu & Run-Min Guo

    2. Department of Physiology and Institute of Neuroscience, Medical College, University of South China, 28 W Changsheng Road, Hengyang, 421001, Hunan, People’s Republic of China

      Jia-Mei Jiang, Hong-Feng Gu, Fan Xiao, Ying Chen & Xiao-Qing Tang

    3. Department of Anthropotomy, Medical College, University of South China, Hengyang, 421001, Hunan, People’s Republic of China

      Li Wang

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    Correspondence to Keng Wu or Xiao-Qing Tang.

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    Jia-Mei Jiang and Li Wang contributed equally to this work.

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    Jiang, JM., Wang, L., Gu, HF. et al. Arecoline Induces Neurotoxicity to PC12 Cells: Involvement in ER Stress and Disturbance of Endogenous H2S Generation. Neurochem Res 41, 2140–2148 (2016). https://doi.org/10.1007/s11064-016-1929-6

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