혈액뇌장벽의 치밀이음부에서 H2O2의 occludin 손상에 대한 알파토코페롤의 억제효과
Alpha-tocopherol Prevents H2O2-induced Tight Junction Occludin Disruption in Blood-Brain Barrier
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
비타민 E 는 사람에게서 지용성 항산화제로 가장 중요하다. 그 중 α-tocopherol은 여러 질병에 보호효과 가 있는 것으로 알려져 있지만, 활성산소종을 내는 질병 중 혈액뇌장벽의 치밀이음부에서 occludin의 변화를 예방 하는지는 알려져 있지 않다. 이 연구에서는 치밀이음부 단백질인 occludin에 대한 H2O2의 영향에 대해 알파토코페롤이 보호작용을 나타내 는 지를 쥐 뇌 미세혈관 내피세포를 일차 배양하여 공초점레이져현미경, Western Blot, cell viability assay로 관찰 하였다. 알파토코페롤은 매우 높은 농도(2.8 mM)에서도 세포독성이 나타나지 않았다. AlamaBlue assay와 위상차현미경 을 통해 관찰한 결과, 알파토코페롤을 전처치하면, H2O2에 의한 세포독성이 억제되었다. 공초점현미경과 western blot을 통해 관찰한 결과, H2O2에 의한 occludin의 소실은 알파토코페롤을 전처치하여 막을 수 있었다. 이와 같은 결과는, 알파토코페롤은 여러 질환으로부터 뇌혈관 내피세포의 보호효과를 나타낼 수 있다는 것을 제시하고 있다. 알파토코페롤은 부작용이 거의 없기 때문에 퇴행성신경질환치료에 사용될 수 있을 것이다.
Abstract
Vitamin E is the most important lipid-soluble antioxidant in humans. Although alpha-tocopherol is suggested that it has protective effect from many diseases, little is known about the prevention of occludin alteration in tight junction of blood-brain barrier (BBB) under pathologic insults producing reactive oxygen species (ROSs). In this study, the effects of alpha-tocopherol on H2O2-induced tight junction occludin were studied. Primary culture of rat brain microvessel endothelial cells was investigated with confocal microscopy, Western blot, and cell viability assay. Alpha-tocopherol had no apparent cytotoxicity up to 2.8 mM. The preincubation with alpha-tocopherol suppressed the H2O2-induced cytotoxicity in Alamar Blue assay and phase contrast microscopy. In confocal laser microscopy and Western blot, H2O2-induced loss of occludin was suppressed by preincubation with alpha-tocopherol. The present findings provide evidence that alpha-tocopherol may be beneficial for cellular protection from pathologic insults. Since alpha-tocopherol was demonstrated to have far fewer adverse effects, it would become a noteworthy nutrient or drug for the treatment of neurodegenerative diseases.
참고문헌 (36)
- "Brain microvessel endothelial cell culture systems", 8, pp.239~258 (1996)
- "Oxidation products andantioxidant markers in plasma of patients with Graves’disease and toxic multinodular goiter effect of methimazoletreatment", 38, pp.659~664 (2004)
- "Induction of drug metabolizing enzymes by vitamin E.", 162, pp.797~802 (2005)
- "Morphology and physiology of theblood-brain barrier", 22, pp.202~214 (2003)
- "Detection of oxidative DNA damage to ischemic reperfusedrat hearts by 8-hydroxydeoxyguanosine formation", 30, pp.1939~1944 (1998)
- "Gene expression in acute myocardial stress. Induction by hypoxia, ischemia, reperfusion, hyperthermia and oxidative stress.", 27, pp.181~193 (1995)
- "Compartmentalized coculture of rat brain endothelial cells and astrocytes: a syngenic model to study the blood-brain barrier", 121, pp.21~31 (2002)
- "Safety of antioxidant vitamins and β-carotene", 62, pp.1510~1516 (1995)
- "Development of the blood-brain barrier", 314, pp.119~129 (2003)
- "Vitamin E therapy in Parkinson’sdisease", 189, pp.129~146 (2003)
- "Hypoxia-induced hyperpermeability in brain microvesselendothelial cells involves VEGF-mediated changes in theexpression of zonula occludens-1", 63, pp.70~80 (2002)
- "Manner of interaction ofheterogeneous claudin species within and between tightjunction strands", 147, pp.891~903 (1999)
- "Vitamin E and Alzheimer disease: the basis for additional clinical trials", 71, pp.630~636 (2000)
- "Occludin asa possible determinant of tight junction permeability inendothelial cells", 110, pp.1603~1613 (1997)
- "Molecular physiology andpathophysiology of tight junctions in the blood-brainbarrier", 24, pp.719~725 (2001)
- "Disruption of epithelial barrier function by H2O2: distinct responses of Caco-2 and Madin-Darby canine kidney (MDCK) strains", 49, pp.101~112 (2003)
- "H2O2 mediated permeability II: importance of tyrosine phosphatase and kinase activity.", 281 (2001)
- "H2O2-mediated permeability: role of MAPK and occludin.", 279 (2000)
- "Antioxidant defense in centenarians (a preliminary study).", 47, pp.281~292 (2000)
- "Hydrogen peroxide-induced alterations of tight junction proteins in bovine brain microvascular endothelial cells", 68, pp.231~238 (2004)
- "Cerebral microvascular changes inpermeability and tight junctions induced by hypoxiareoxygenation", 282 (2002)
- "Vitamin E modulation of cardiovascular disease", 1031, pp.271~279 (2004)
- "Vitamin E modulation of cardiovascular disease", 1031, pp.271~279 (2004)
- "Assessment of the safety of supplementation with different amo unts of vitamin E in healthy older adults", 68, pp.311~318 (1998)
- "Reassembly of the tight junction after oxidative stress depends on tyrosine kinase activity", 276, pp.22048~22055 (2001)
- "Regulation of geneexpression by oxygen: NF-B and HIF-1, two extremes.", 33, pp.1231~1242 (2002)
- "Escape from redox regulation enhances the transformingactivity of Fos", pp.695~8 701 (1993)
- "Vitamin E protects porcine but notbovine cultured aortic endothelial cells from oxygen-derivedfree radical injury due to hydrogen peroxide", 20, pp.55~67 (2004)
- "Changes in cytoskeletal and tight junctional proteins correlate with decreased permeability induced by dexamethasone in cultured rat brain endothelial cells", 344, pp.112~116 (2003)
- "The cell biology of the blood-brainbarrier", 22, pp.11~18 (1999)
- "an oxidative stress-responsive transcription factor of eukaryoticcells", pp.221~237 (1992)
- "Anti-inflammatory effects of alphatocopherol", 1031, pp.195~203 (2004)
- "Reactive oxygen species in cellsignaling", 279 (2000)
- "Impact of deprenyl and toco-pherol treatment on Parkinson’s disease in DATATOPpatients requiring levodopa", 39, pp.37~45 (1996)
- "Occludin and claudin in tight junctionstrands leading or supporting players?", 9, pp.268~273 (1999)
- "Changes of oxidative stress parameters in diabetic pregnancy", 38, pp.795~803 (2004)