본문 바로가기
KERD
논문KCI 등재

혈청결핍 상태에서 크레아틴이 RGC-5세포의 생존에 미치는 영향

Effect of Creatine on the Survival of RGC-5 Cells under Serum Deprivation

김윤영|강선희|김재우|홍정흠
대한안과학회 (The Korean Ophthalmological Society)|2011.05|대한안과학회지 (Journal of The Korean Ophthalmological Society)|제52권 제5호|pp. 618-623

0

조회

0

다운로드

0

인용

0

참고문헌

키워드

CreatineRetinal ganglion cellCytoprotection
주제분류안과학

초록

목적: 혈청결핍 상태에서 세포의 에너지 전구물질로 작용하는 크레아틴이 RGC-5세포의 생존에 미치는 영향을 알아 보고자 하였다. 대상과 방법: 혈청이 포함되지 않은 배지에서 RGC-5세포를 5 mM의 크레아틴에 노출시킨 후 세포의 생존을 MTT assay로 측정하였으 며, 미토콘드리아의 호흡활성도를 resazurin assay로 조사하였다. 세포고사의 정도를 알아보기 위해 acridine orange/Hoechest 33342로 생체염색을 시행하였으며 annexin/PI 이중염색을 이용한 유세포분석을 시행하였다. 결과: 크레아틴은 혈청결핍상태에서 RGC-5세포의 생존을 유의하게 증가시켰으며, 생체염색과 유세포분석에서 세포고사를 감소시켰 다. 이러한 효과는 미토콘드리아의 호흡활성의 증가와 동반되어 나타났다. 결론: 에너지 전구물질인 크레아틴은 혈청결핍상태에서 세포의 생존을 증가시켰으며 이는 크레아틴이 망막신경절세포의 보호할 가능 성이 있음을 보여 준다.

Abstract

Purpose: To evaluate the protective effect of creatine on the survival of retinal ganglion cells after serum deprivation. Methods: RGC-5 cells were exposed to 5 mM creatine with serum-free media for 4 days. Cellular survival and mitochondrial respiratory activity were measured with MTT assay and resazurin assay, respectively. Degree of apoptosis was evaluated with vital staining using acridine orange/Hoechest 33342 and flow cytometric analysis using annexin/PI, respectively. Results: Creatine increased cellular survival of RGC-5 cells significantly after serum deprivation. Additionally, creatine increased mitochondrial respiratory activity and inhibited apoptosis of RGC-5 cells. Conclusions: The energy precursor creatine increased survival of retinal ganglion cells after serum deprivation. Creatine could be relevant for the cytoprotection of retinal ganglion cells.

참고문헌 (36)

  1. "CNS energy metabolism as related to function", Brain Res Brain Res Rev, 34, pp.42~68 (2000)
  2. Holtzman D, "Brain cellular and mitochondrial respiration in media of altered pH", Metab Brain Dis, 2, pp.127~137 (1987)
  3. Hemmer W, "Functional aspects of creatine kinase in brain", Dev Neurosci, 15, pp.249~260 (1993)
  4. Wallimann T, "Subcellular compartmentation of creatine kinase isoenzymes, regulation of CK and octameric structure of mitochondrial CK", Prog Clin Biol Res, 315, pp.159~176 (1989)
  5. Wallimann T, "Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands", Biochem J, 281, pp.21~40 (1992)
  6. McNaught KS, "Isoquinoline derivatives as endogenous neurotoxins in the aetiology of Parkinson’s disease", Biochem Pharmacol, 56, pp.921~933 (1998)
  7. Brewer GJ, "Protective effect of the energy precursor creatine against toxicity of glutamate and beta-amyloid in rat hippocampal neurons", J Neurochem, 74, pp.1968~1978 (2000)
  8. Michaelis T, "Proton MRS of oral creatine supplementation in rats", NMR Biomed, 12, pp.309~314 (1999)
  9. Wilken B, "Creatine protects the central respiratory network of mammals under anoxic conditions", Pediatr Res, 43, pp.8~14 (1998)
  10. Wilken B, "Anoxic ATP depletion in neonatal mice brainstem is prevented by creatine supplementation", Arch Arch Dis Child Fetal Neonatal Ed, 82, pp.224~227 (2000)
  11. Zhu S, "Prophylactic creatine administration mediates neuroprotection in cerebral ischemia in mice", J Neurosci, 24, pp.5909~5912 (2004)
  12. Adcock KH, "Neuroprotection of creatine supplementation in neonatal rats with transient cerebral hypoxia ischemia", Dev Neurosci, 24, pp.382~388 (2002)
  13. Chen L, "Expression of brain type creatine kinase and ubiquitous mitochondrial creatine kinase in the fetal rat brain", J Comp Neurol, 363, pp.389~401 (1995)
  14. Hemmer W, "Brain type creatine kinase in photoreceptor cell outer segments", J Cell Sci, 106, pp.671~683 (1993)
  15. Dolder M, "Inhibition of the mitochondrial permeability transition by creatine kinase substrates. Requirement for microcompartmentation", J Biol Chem, 278, pp.17760~17766 (2003)
  16. O'Gorman E, "The role of creatine kinase in inhibition of mitochondrial permeability transition", FEBS Lett, 414, pp.253~257 (1997)
  17. Acosta ML, "Creatine transporter localization in developing and adult retina", Am J Physiol Cell Physiol, 289, pp.1015~1023 (2005)
  18. Krishnamoorthy RR, "Characterization of a transformed rat retinal ganglion cell line", Brain Res Mol Brain Res, 86, pp.1~12 (2001)
  19. Hansen MB, "Re examination and further development of a precise and rapid dye method for measuring cell growth cell kill", J Immunol Methods, 119, pp.203~210 (1989)
  20. Mpoke SS, "Differential staining of apoptotic nuclei in living cells application to macronuclear elimination in Tetrahymena", J Histochem Cytochem, 45, pp.675~683 (1997)
  21. Vermes I, "A novel assay for apoptosis,In Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V", J Immunol Methods, 184, pp.39~51 (1995)
  22. Osborne NN, "Pathogenesis of ganglion cell death in glaucoma and neuroprotection", Prog Brain Res, 173, pp.339~352 (2008)
  23. Abu Amero KK, "Detection of mitochondrial respiratory dysfunction in circulating lymphocytes using resazurin", Arch Pathol Lab Med, 129, pp.1295~1298 (2005)
  24. Wallimann T, "Creatine kinase in non muscle tissues and cells", Mol Cell Biochem, 133, pp.193~220 (1994)
  25. Wallimann T, "Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands", Biochem J, 281, pp.21~40 (1992)
  26. Whittingham TS, "Cerebral synaptic transmission during anoxia is protected by creatine", J Neurochem, 37, pp.1618~1621 (1981)
  27. Carter AJ, "Preincubation with creatine enhances levels of creatine phosphate and prevents anoxic damage in rat hippocampal slices", J Neurochem, 64, pp.2691~2699 (1995)
  28. Chung I, "Attenuation of hypoxic current by intracellular applications of ATP regenerating agents in hippocampal CA1 neurons of rat brain slices", Neuroscience, 86, pp.1101~1107 (1998)
  29. Lemasters JJ, "Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death", J Bioenerg Biomembr, 31, pp.305~319 (1999)
  30. O'Gorman E, "The role of creatine kinase in inhibition of mitochondrial permeability transition", FEBS Lett, 414, pp.253~257 (1997)
  31. Wyss M, "Creatine and creatinine metabolism", Physiol Rev, 80, pp.1107~1213 (2000)
  32. Wyss M, "Mitochondrial creatine kinase: a key enzyme of aerobic energy metabolism", Biochim Biophys Acta, 1102, pp.119~166 (1992)
  33. Passonneau JV, "Metabolite flux in single neurons during ischemia and anesthesia", Curr Probl Clin Biochem, 3, pp.198~212 (1971)
  34. Stachowiak O, "Mitochondrial creatine kinase is a prime target of peroxynitrite-induced modification and inactivation", J Biol Chem, 273, pp.16694~16699 (1998)
  35. Ganapathy PS, "Sensitivity of staurosporine induced differentiated RGC 5 cells to homocysteine", Curr Eye Res, 35, pp.80~90 (2010)
  36. Wood JP, "A comparison of differentiation protocols for RGC 5 cells", Invest Ophthalmol Vis Sci, 51, pp.3774~3783 (2010)

관련 논문AI추천

R28세포에서 고포도당이 반응성산소종의 생성에 미치는 영향김재우, 이종민대한안과학회 · 대한안과학회지2010 · 제51권 제5호산화스트레스가 유도된 망막색소상피세포에서 폴리페놀 화합물이 보이는 세포보호 효과곽형우, 서경훈, 유승영대한안과학회 · 대한안과학회지2016 · 제57권 제1호Effect of Anti-vascular Endothelial Growth Factor Antibody on the Survival of Cultured Retinal Ganglion Cells김찬윤, 배형원, 성공제, 이상엽, 이지민대한안과학회 · Korean Journal of Ophthalmology2017 · 제31권 제4호Genistein이 섬유주세포의 생존과 일산화질소의 생성에 미치는 영향김윤영, 김재우, 홍정흠대한안과학회 · 대한안과학회지2011 · 제52권 제8호망막신경절세포에서 고장성 및 고오스몰 스트레스에 의한 긴장성반응유도 결합단백질의 발현권민영, 우제문, 우종은, 정수월대한안과학회 · 대한안과학회지2014 · 제55권 제8호심바스타틴이 인체망막색소상피세포에서 생산된 혈관내피성장인자에 미치는 효과김경실, 김경진, 김나래, 진희승대한안과학회 · 대한안과학회지2012 · 제53권 제6호

이 간행물의 다른 논문 (19건)

브리모니딘 분무와 관류에 따른 눈물배출계의 형태학적 변화오동은, 양현승pp. 511-518코경유 눈물주머니코안연결술의 학습곡선구병영, 손준혁, 김원제pp. 519-523각막절편 두께 측정법의 비교: 초음파 각막두께측정계의 차감법, 직접측정법과 빛간섭단층촬영계이주은, 김명준, 김재용, 차흥원, 이수환pp. 524-530당뇨망막병증 검진에 있어 무산동 디지털 안저사진기의 효용성배정훈, 송수정, 이대웅pp. 531-536원발성열공망막박리 환자에서 단독유리체절제술과 수정체유화술 및 유리체절제술병합술의 결과 비교장우혁, 구병영, 사공민pp. 537-543
원문 보기