대한한방내과학회지 (The Journal of Internal Korean Medicine)

대한한방내과학회 (The Society of Internal Korean Medicine)
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H2O2로 유발된 C6 신경교세포 사멸에 대한 총명공진단의 보호 효과

Protective Effects of Chongmyunggongjin-dan on H2O2-induced C6 Glial Cell Death

  • 황규상 (원광대학교 한의과대학 내과학교실) ;
  • 신용진 (원광대학교 한의과대학 내과학교실)
  • Hwang, Gyu-sang (Dept. of Internal Medicine, College of Korean Medicine, Wonkwang University) ;
  • Shin, Yong-jeen (Dept. of Internal Medicine, College of Korean Medicine, Wonkwang University)
  • 투고 : 2020.02.24
  • 심사 : 2020.03.30
  • 발행 : 2020.04.20
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초록

Objectives: This study was conducted to identify the protective effects of Chongmyunggongjin-dan (CMGJD) on Hydrogen peroxide (H2O2)-induced apoptosis mechanisms in C6 glial cells. Method: We used CMGJD after distilled water extraction, filtration, and lyophilization. The ROS scavenging effect was examined by fluorescence microscopy. Expression levels of proteins related to ROS generation were investigated by western blotting. Functional changes in organelles related to Reactive oxygen species (ROS) generation were investigated by immunoblotting and by verifying expression level of relevant enzymes. Results: The CMGJD extract protected the cells against H2O2-induced morphological changes and DNA fragmentation, inhibited the increase of Heme_oxygenase-1(HO-1) and the decrease in catalase, protected against the loss of mitochondrial membrane potential, inhibited disturbances of lysosomal function, and induced an increase in peroxisomes. Conclusion: CMGJD was confirmed to have a protective effect on H2O2-induced C6 glial cell death possibly by blocking the pathways causing damage to subcellular organelles, such as mitochondria, lysosomes, and peroxisomes. We assume that CMGJD will be effective for the prevention and treatment of ischemic stroke in a clinical environment.

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참고문헌

  1. 전국 한의과대학 심계내과학 교수협의회. 한방 순환.신경내과학 신경계 편. 서울: 우리의학서적; 2016, p. 80-4.
  2. Jameson JL, Kasper DL, Longo DL, Fauci AS, Hauser SL, Loscalzo J. Harrison's principles of internal medicine. 20th edition. Vol 2. New York: McGraw-Hill Education; 2018, p. 3079-80.
  3. Lipton P. Ischemic Cell Death in Brain Neurons. Physiol Review 1999;79(4):1499. https://doi.org/10.1152/physrev.1999.79.4.1431
  4. Iwata-Ichikawa E, Kondo Y, Miyazaki I, Asanuma M, Ogawa N. Glial Cells Protect Neurons Against Oxidative Stress via Transcriptional Up-Regulation of the Glutathione Synthesis. Journal of Neurochemistry 1999;72(6):2334-44. https://doi.org/10.1046/j.1471-4159.1999.0722334.x
  5. 王璆. 是齋百一選方. 상해: 상해과학기술출판사; 1991, p. 280.
  6. 許浚. 東醫寶鑑. 서울: 여강출판사; 2005, p. 124, 1750.
  7. 危亦林. 世醫得效方. 북경: 인민위생출판사; 1990, p. 142.
  8. Choi KH, Park CS. An analysis of the Gongjindan's ingredients and its efficacy on anti-oxidation. Kor J Herbology 2007;22(2):51-63.
  9. Jung JW, Jeon SH, Bae WJ, Kim SJ, Chung MS, Yoon BI, et al. Suppression of Oxidative Stress of Modified Gongjin-Dan (WSY-1075) in Detrusor Underactivity Rat Model Bladder Outlet Induced by Obstruction. Chin J Integr Med 2018;24(9):670-5. https://doi.org/10.1007/s11655-017-2970-1
  10. Kim HJ, Kim YS, Mok JY, Jeong SI, Hwang SY, Cho JK, et al. Anti-inflammatory Effect of Gagam-GongJin-dan in mouse peritoneal macrophages. The Korean Journal of oriental medical prescription 2011;19(1):207-17.
  11. Sunwoo YY, Park SI, Chung YA, Lee J, Park MS, Jang KS, et al. A pilot study for the neuroprotective effect of Gongjin-dan on transient middle cerebral artery occlusion-induced ischemic rat brain. Evid Based Complement Alternat Med 2012;2012:682-720.
  12. Joo JJ, Kim YS, Lee JD, Choi DY, Koh HK, Ahn BC, et al. The Effect of Kongjindan Aqua-acupuncture on the CD4+ T cell count at al. in rats were administered with MTX. Journal of Korean Acupuncture & Moxibustion Society 1999;16(3):179-202.
  13. Jang SI, Kim HJ, Mok JY, Park KH, Jeong SI, Hwang BS, et al. Hepatoprotective Effect of Gagam-GongJin-dan extract against Acetaminophen-Induced Liver Injury in Mice. The Korea Journal of Herbology 2010;25(3):149-57. https://doi.org/10.6116/KJH.2010.25.3.149
  14. Hur H, Kim HJ, Park K, Kwak MA, Kim DJ, Byun JS. Hepatoprotective effects of Gongjin-dan on ethanol-mediated experimental liver damage in rats. The Journal of Internal Korean Medicine 2008;29(4):1037-47.
  15. Seong KM, Hae RK, Son BK. The Effect of Gongjin-dan on Gliosis in Middle Cerebral Artery Occlusion (MCAO) Rats. The Journal of Internal Korean Medicine 2009;30(4):674-84.
  16. Hwang SM, Chung DK. The Effects of KongJin-Dan(KJD) on the Alzheimer's Disease Model Induced by CT105. The Korean Society of Oriental Neuropsychiatry 2004;15(2):103-18.
  17. Son MJ, Im HJ, Ku B, Lee JH, Jung SY, Kim YE, et al. An Herbal Drug, Gongjin-dan, Ameliorates Acute Fatigue Caused by Short-Term Sleep-Deprivation: A Randomized, Double-Blinded, Placebo-Controlled, Crossover Clinical Trial. Front Pharmacol 2018;11(9):479.
  18. Lee JM, Cho JH, Jang JB, Lee KS. Effects of Yangyogongjindan on the Reproductive Competence of Aged Mice. Journal of Oriental Obstetrics & Gynecology 2004;17(1):57-71.
  19. Kim KH, Kim YH, Son JE, Lee JH, Kim S, Choe MS, et al. Intermittent fasting promotes adipose thermogenesis and metabolic homeostasis via VEGF-mediated alternative activation of macrophage. Cell Res 2017;27(11):1309-26. https://doi.org/10.1038/cr.2017.126
  20. Astrup J, Siesjo BK, Symon L. Thresholds in cerebral ischemia-The ischemic penumbra. Stroke 1981;12(6):723-5. https://doi.org/10.1161/01.str.12.6.723
  21. Kim DH, Kim SM, Cho HG, Cha YS, Heo Y, Moon BS, et al. Effects of Samul-tang on Nitric Oxide Induced-cytotoxicity in C6 Glial Cell. The Journal of Internal Korean Medicine 2000;21(4):535-42.
  22. Cho NS, Rhyu JK, Lee I, Shin SH, Moon BS, Na YH. Effects of Jagamcho-tang on the C6 Glial Cell Injured by LPS Combined PMA. The Journal of Internal Korean Medicine 2000;21(3):467-76.
  23. Lee YG, Moon BS. Protective Effects of Ukyium (右歸飮, Yougui-yin) in Zinc-induced Apoptosis of C6 Glial Cells. Journal of Korean Medicine 2001;22(3):63-73.
  24. Solary E, Droin N, Bettaieb A, Corcos L, Dimanche-Boitrel MT, Garrido C. Positive and negative regulation of apoptotic pathways by cytotoxic agents in hematological malignancies. Leukemia 2000;14(10):1833-49. https://doi.org/10.1038/sj/leu/2401902
  25. Li Z, Theus MH, Wei L. Role of ERK 1/2 signaling in neuronal differentiation of cultured embryonic stem cells. Develop Growth Differ 2006;48(8):513-23. https://doi.org/10.1111/j.1440-169X.2006.00889.x
  26. Halliwell B, Gutteridge. Protection aganist oxidants in biological systems: the superoxide theory of oxygen toxicity. In Free radicals in biology and medicine. 2nd Ed. Oxford: Clarendon Press; 1999, p. 86-9.
  27. Fang J, Akaike T, Maeda H. Antiapoptotic role of heme oxygenase (HO) and the potential of HO as a target in anticancer treatment. Apoptosis 2004;9(1):27-35. https://doi.org/10.1023/b:appt.0000012119.83734.4e
  28. 黃度淵. 對譯 證脈.方藥合編. 서울: 남산당; 2007, p. 163-4.
  29. 신민교. 임상본초학. 파주: 영림사; 2006, p. 188, 248, 254, 474.
  30. Lee JH. A Literature Review and Research Evaluation of Chongmyungtang. Department of Alternative Medicine, Graduate School of Health Science, Chosun University 2012.
  31. 권학철, 배나영, 양현옥, 오명숙, 정성권. 용안육 추출물 또는 이를 포함하는 혼합 추출물을 함유하는 퇴행성 뇌신경질환의 예방 또는 치료용 조성물. 서울: 한국과학기술연구원; 2010, p. 1-26.
  32. Morgan MJ, Kim YS, Liu Z. Lipid rafts and oxidative stress-induced cell death. Antioxid Redox Signal 2007;9(9):1471-83. https://doi.org/10.1089/ars.2007.1658
  33. Zorova LD, Popkov VA, Plotnikov EY, Silachev DN, Pevzner IB, Jankauskas SS, et al. Mitochondrial membrane potential. Anal Biochem 2018;552(1):50-9. https://doi.org/10.1016/j.ab.2017.07.009
  34. Kobayashi S. Choose Delicately and Reuse Adequately: The Newly Revealed Process of Autophagy. Biological & Pharmaceutical Bulletin 2015;38(8):1098-103. https://doi.org/10.1248/bpb.b15-00096
  35. Esteves AR, Arduino DM, Silva DF, Oliveira CR, Cardoso SM. Mitochondrial Dysfunction: The Road to Alpha-Synuclein Oligomerization in PD. Parkinson's Disease 2011;693761.
  36. Chen F, Deng Z, Xiong Z, Zhang B, Yang J, Hu J. A ROS-mediated lysosomal-mitochondrial pathway is induced by ginsenoside Rh2 in hepatoma HepG2 cells. Food Funct 2015;6(12):3828-37. https://doi.org/10.1039/C5FO00518C
  37. Chen S, Zhou C, Yu H, Tao L, An Y, Zhang X, et al. 27-Hydroxycholesterol Contributes to Lysosomal Membrane Permeabilization-Mediated Pyroptosis in Co-cultured SH-SY5Y Cells and C6 Cells. Front Mol Neurosci 2019;12:14. https://doi.org/10.3389/fnmol.2019.00014
  38. Park S, Choi SG, Yoo SM, Son JH, Jung YK. Choline dehydrogenase interacts with SQSTM1/p62 to recruit LC3 and stimulate mitophagy. Autophagy 2014;10(11):1906-20. https://doi.org/10.4161/auto.32177
  39. Lodhi IJ, Semenkovich CF. Peroxisomes: a nexus for lipid metabolism and cellular signaling. Cell Metab 2014;19(3):380-92. https://doi.org/10.1016/j.cmet.2014.01.002