Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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The Protective Effect of Lonicerae flos Extract on Cultured C6 Glioma Cells Damaged by Aluminum of Dementia Inducer

치매유발제인 알루미늄으로 손상된 배양 C6 Glioma 세포에 대한 금은화 추출물의 보호 효과

  • Jung, Jai-Yun (Department of Anesthesiology and Pain Medicine, Sanbon Hospital, Wonkwang University College of Medicine) ;
  • Jung, In-Ju (Department of Cosmetology, Dongshin University) ;
  • Jekal, Seung-Joo (Department of Clinical Laboratory Science, Wonkwang Health Science University)
  • 정재윤 (원광대학교 의과대학 산본병원 마취통증의학과) ;
  • 정인주 (동신대학교 뷰티미용학과) ;
  • 제갈승주 (원광보건대학교 임상병리학과)
  • Received : 2017.07.31
  • Accepted : 2017.08.28
  • Published : 2017.09.30
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Abstract

This study was performed to evaluate the cytotoxicity of aluminum chloride ($AlCl_3$), and the protective effect of Lonicerae flos (LF) extract on $AlCl_3-induced$ cytotoxicity in the cultured C6 glioma cells. Here, the cell viability and antioxidative effects, such as DPPH-radical scavenging activity, superoxide anion scavenging activity, and lipid peroxidation (LP), were assessed. $AlCl_3$ significantly decreased the cell viability in a dose-dependent manner; the $XTT_{50}$ value was measured at $128.8{\mu}M$ of $AlCl_3$. The cytotoxicity of $AlCl_3$ was determined as highly toxic the y Borenfreund and Puerner's toxic criteria. The butylated hydroxytoluene (BHT) and antioxidant both significantly increased the cell viability, which was damaged by $AlCl_3-induced$ cytotoxicity in these cultures. In the protective effect of LF extract on $AlCl_3-induced$ cytotoxicity, the LF extract significantly increased the superoxide anion scavenging activity and inhibitory activity of LP, as well as the DPPH-radical scavenging activity. From these results, it is suggested that the oxidative stress may have been involved in the cytotoxicity of $AlCl_3$, and LP extract effectively protected $AlCl_3-induced$ cytotoxicity through the antioxidative effects. Conclusively, the natural resources, like LP extract, may be a putative therapeutic agent for the treatment of dementia induced by allergen like aluminum correlated with the oxidative stress.

치매유발제인 염화알루미늄(aluminum chloride, $AlCl_3$)의 세포독성을 대뇌교종세포인 C6 glioma 세포를 대상으로 산화적 손상측면에서 조사하였으며, 또한 $AlCl_3$의 세포독성에 대한 금은화(Lonicerae flos, LF) 추출물의 영향을 세포생존율을 비롯한 DPPH-라디칼 소거능, superoxide anion-radical (SAR) 제거능 및 지질과산화(lipid peroxidation, LP) 억제능과 같은 항산화 측면에서 분석하였다. 본 실험에서 배양 C6 glioma 세포에 $120{\sim}160{\mu}M$의 $AlCl_3$가 각각 포함된 배양액에서 처리한 결과 처리 농도에 비례하여 세포생존율이 대조군에 비하여 유의한 감소를 보였으며, 이 때 $XTT_{50}$값은 $128.8{\mu}M$에서 나타나 중간독성(mid-toxic)인 것으로 나타났다. 또한, 항산화제인 BHT는 $AlCl_3$의 세포독성을 유효하게 방어하였다. 한편, $AlCl_3$의 세포독성에 대한 금은화(LF) 추출물의 영향에 대한 조사에 있어서, LF 추출물은 $AlCl_3$에 의하여 감소된 세포생존율을 유의하게 증가시킴으로써 세포독성을 방어하였다. 이와 동시에, LF 추출물은 DPPH-라디칼 소거능을 비롯하여 SAR 제거능 및 LP 억제능을 보임으로서 항산화 효과를 나타냈다. 위의 결과로부터 $AlCl_3$의 세포독성에 산화적 손상이 관여하고 있으며, 또한 LF 추출물은 $AlCl_3$의 독성을 항산화능에 의하여 효과적으로 방어하였다. 따라서, LF 추출물과 같은 천연성분은 알루미늄과 같이 산화적 손상과 관련된 치매유발제의 독성을 방어함으로써 차후 치매와 같은 산화적 손상 관련의 질환 치료를 위한 항산화 물질로서의 개발적 가치가 있다고 사료된다.

Keywords

References

  1. Ellis JM. Cholinesterase inhibitors in the treatment of dementia. J Am Osteopath Assoc. 2005;105(13):145-158.
  2. Kim SJ, Yu YW, Lee JK. Cytotoxicity and protective effect of Portulaca oleracea L. extract on cultured neuroglioma cells damaged by aluminum of dementia-induced agent. J Kor Soc Plants Environ. 2013;16(5):251-256. https://doi.org/10.11628/ksppe.2013.16.5.251
  3. Kourie JI. Mechanisms of amyloid beta protein-induced modification in ion transfort systems; implications for neurodegenerative disease. Cell Mol Neurobiol. 2001;21(3):173-213. https://doi.org/10.1023/A:1010932603406
  4. Kim SC, Lee JR, Choi KI, Park ST, Kwon YK, Byun SH. Effect of Lonicerae flos on contact hypersensitivity induced by repeat elicitation of DNCB. Kor J Herbol. 2006;21(1):9-15.
  5. Bryan-Sisneros AA, Fraser SP, Suh YH, Djamgoz MB. Toxic effect of the beta-amyloid precursor protein C-terminus fragment and Na+/$Ca^{2+}$gradients. Neuroreport. 2000;11(15):3357-3360. https://doi.org/10.1097/00001756-200010200-00019
  6. Stephen LY, Loyd HB, June KA, Joyce MA, Michael DD, Paula BE, et al. Amyloid ${\beta}$ and amylin fibrils induce increase in proinflammatory cytokine and chemokine production by THP-1 cells and murine microglia. J Neurochem. 2000;74(3):1017-1025. https://doi.org/10.1046/j.1471-4159.2000.0741017.x
  7. Kontush A. Amyloid-beta ; an antioxidant that becomes a pro-oxidant and critically contributes to Alzheimer's disease. Free Radic Bio Med. 2001;31(19):1120-1131. https://doi.org/10.1016/S0891-5849(01)00688-8
  8. Kim YJ. Etiology and pathogenesis of Parkinson's disease. J Kor Neurol Assoc. 2004;22(5):421-432.
  9. Roger DG, Olivier F, Clank P, Courtosis M, Piriou A. Aluninum transfer as complex through blood-brain barrier. Biological Rrace Element Res. 1990;25(5):39-45. https://doi.org/10.1007/BF02990262
  10. Son YW, Choi YS. Protective of Chenopodium album var. exract by oxidative effect in cultured human skin fibroblasts damaged by cadmium. J Kor Soc People plants Environ. 2012;15(3): 155-161.
  11. Li YL, Gan GP, Zhang HZ, Wu HZ, Li CL, Hung YP, et al. A flavonoid glycoside isolated from Smilax china L. rhizome in vitro anticancer effects on human cancer cell lines. J Ethnopharmacol. 2007;113(1):115-124. https://doi.org/10.1016/j.jep.2007.05.016
  12. Choi CW, Jung HA, Kang SS, Choi JS. Antioxidant constituents and a new triterpenoid glycoside from Lonicerae flos. Arch Pharm Res. 2007;30(1):1-7. https://doi.org/10.1007/BF02977770
  13. Kwon SH, Kim HC, Lee SY, Jang CG. Loganin improves learning and memory impairments induced by scopolamine in mice. Eur J Pharmacol. 2009;619(1):44-49. https://doi.org/10.1016/j.ejphar.2009.06.062
  14. Kim TY, Jekal SJ. Antioxidative effect of Chelidonium majus extract on cultured NIH3T3 fibroblasts injured by cadmium chloride of toxicant. Kor J Clin Lab Sci. 2016;48(1):1-7. https://doi.org/10.15324/kjcls.2016.48.1.1
  15. Han DS, Baeck KH, Kim YO, Choi KE, Kwag JS, Baeck SH. Development of anticancer agents from Korean medicinal plants. Part 6. Cytotoxic activity of the ethyl acetate soluble fraction of Lonicerae flos against human oral epithelioid carcinoma cells. Kor J Pharmacogn. 1998;29(1):22-27.
  16. Zhang X, Guo Q, Yu B. Rapid quantitative analysis of adulterant Lonicera species in preparations of Lonicerae Japonicae flos. J Sep Sci. 2015;38(23):4014-4020. https://doi.org/10.1002/jssc.201500917
  17. Park ST, Lim KT, Chung YT, Kim SU. Methymercury-induced neurotoxicity in cerebral neuron culture is blocked by anti-oxidants and NMDA receptor antagonists. Neurotoxicol. 1996; 17(1):37-46.
  18. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth. 1983;65(1-2):55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  19. Blois MS. Antioxidant determination by the use of a stable free radical. Nature. 1958;181(4617):1199-1200. https://doi.org/10.1038/1811199a0
  20. Kikuzaki H. Nakatani N. antioxidant effects of some ginger constituents. J Food Sci. 1993;58(6):1407-1410. https://doi.org/10.1111/j.1365-2621.1993.tb06194.x
  21. Borenfreund E, Puerner JA. A simple quantitative procedure using monolayer culture for cytotoxicity assay (HTD/NR-90). J Tiss Cult Meth. 1984;9(1):7-9. https://doi.org/10.1007/BF01666038
  22. Verstraeten SV, Amio L, Oteiza PI. Aluminum and lead molecular mechanisms of brain toxicity. Arch Toxicol. 2008;82(11): 789-802. https://doi.org/10.1007/s00204-008-0345-3
  23. Deloncle R, Guillard O. Mechanism of Alzheimer's disease: arguments for a nerotransmitter-aluminum complex implication. Neurochem Research. 1990;15(12):1239-1245. https://doi.org/10.1007/BF01208586
  24. Waltz JA, Knowlton BJ, Holyoak KJ, Boone KB, Back-Madruga C, McPherson S, et al. Relational integration and executive function in Alzheimer's disease. Neuropsychology. 2004;18(2): 296-302. https://doi.org/10.1037/0894-4105.18.2.296
  25. Kim SE, Yang BS, Choi YS. Cytotoxicity and effect of Lonicerae flos extract against chromium, contact dermatitis-induced agent in cultured human skin fibroblasts. J Kor Soc People Plants Environ. 2012;15(6):407-412. https://doi.org/10.11628/ksppe.2012.15.6.407
  26. Kwak WJ, Han CK, Chang HW, Kim HP, Kang SS, Son KH. Loniceroside C , an antiinflammatory saponin from Lonicerae japonica. Chem Pharm Bull. 2003;51(3):333-335. https://doi.org/10.1248/cpb.51.333
  27. Ma J, Luo XD, Protiva P, Tang H, Ma C, Basile MJ, et al. Bioactive novel polyphenols from the fruit of Manikara zapota (Sapodolla). J Nat Prod. 2003;66(7):983-986. https://doi.org/10.1021/np020576x
  28. Rim BM, Rim CW, Choi YJ, Chung YS, Jeong HG. Effects of Lonicerae japonica extract as a biological response modifier. Environ Mut Car. 1992;12(1):45-54.
  29. Oh YL, Choi YR, Chang BS, Jung IJ. Antioxidative effect of Portulaca oleracea L. extract on allergic contact dermatitis agent, copper in cultured human skin fibroblasts. J Invest Cosm. 2012;8(4):243-249. https://doi.org/10.15810/jic.2012.8.4.004

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