36 research outputs found
λ§λ§μμμνΌμΈν¬μ£Όμ μ°νμ€νΈλ μ€ μ λ° λͺ¨λΈμμ μμ€μ½λ₯΄λΈμ°κ³Ό μμ€νμν΄μ νμ°νν¨κ³Όμ κ΄ν μ°κ΅¬
νμλ
Όλ¬Έ(μμ¬)--μμΈλνκ΅ λνμ :μκ³Όλν νλκ³Όμ μ€κΈ°μΈν¬μλ¬Όνμ 곡,2019. 8. μ νκ³€.Purpose: This study is to research antioxidative effect of ascorbic acid (vitamin C) and astaxanthin on ARPE-19 cells within an oxidative stress model induced by endogenous (hydrogen peroxide) and exogenous (ultraviolet B) sources of reactive oxygen species (ROS).
Methods: 0, 0.1, 0.2, 0.4, 0.6, 0.8mM of hydrogen peroxide and 0, 20, 40, 60, 80, 100mJ/cm2 of UVB were treated to ARPE-19 cells to find sublethal and lethal dose of each source. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and 2β²,7β²-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay were done to examine cell viability and intracellular reactive oxygen species level change. With the sublethal and lethal dose of each inducers, 0-750uM of ascorbic acid and 0-40uM of astaxanthin were treated to examine their antioxidative effect on the oxidative stress induced ARPE-19.
Results: Ascorbic acid and astaxanthin had antioxidative effects on ARPE-19 cells by significantly increasing cell viability and reducing intracellular reactive oxygen species level after oxidative stress induction. 500uM ascorbic acid increased the cell viability 27% and 14% respectively for 0.2mM and 0.4mM hydrogen peroxide and 17% and 12% respectively for 20mJ/cm2 and 100mJ/cm2 of UVB. The increment was due to its antioxidative effect considering decreased intracellular ROS level. Astaxanthin also showed an antioxidative effect but extended time period of time was needed for it to act. 30 hours of culture with 90uM ascorbic acid and 20uM astaxanthin increased the cell viability from 75% to 129% when exposed to 0.2mM hydrogen peroxide.
Conclusion: Our data suggest that hydrogen peroxide and UVB-induced oxidative stress model of ARPE-19 could be used to examine antioxidative effect of compounds of interest. Ascorbic acid and astaxanthin can be strong antioxidants and the mixture of the two compounds can have synergistic effect.λͺ©μ : κ³Όμ°νμμλ‘ μΈν¬ λ΄λΆμμ λ°μν μ°νμ€νΈλ μ€λ₯Ό μ¬ννκ³ μ€νμ₯ μμΈμ μΌλ‘ μΈν¬ μΈλΆμ μ°νμ€νΈλ μ€λ₯Ό μ¬νν λͺ¨λΈμ μ¬μ©νμ¬ μμ€μ½λ₯΄λΈμ°κ³Ό μμ€νμν΄μ΄ λ§λ§μμμνΌμΈν¬μ νμ°ν ν¨κ³Όκ° μλμ§ μμλ³΄κ³ μ νμλ€.
λ°©λ²: 0, 0.1, 0.2, 0.4, 0.6, 0.8mMμ λλμ κ³Όμ°νμμμ 0, 20, 40, 60, 80, 100mJ/cm2 μΈκΈ°μ μ€νμ₯ μμΈμ μ μ²λ¦¬νμ¬ μΈν¬μ μμ‘΄μ¨κ³Ό μΈν¬ λ΄μ νμ±μ°μ’
μ λλ λ³νλ₯Ό μΈ‘μ νμλ€. μΈν¬μ μμ‘΄λ₯ μ MTT assay, μΈν¬ λ΄μ νμ±μ°μμ’
μ λλλ DCFH-DA assayλ₯Ό ν΅ν΄μ μΈ‘μ νμλ€. λ μ°νμ€νΈλ μ€ μ λ°μΈμμ μΉμ¬λ, μ€μΉμ¬λμ 쑰건μμ 0-750uM λλμ μμ€μ½λ₯΄λΈμ°κ³Ό 0-40uM λλμ μμ€νμν΄μ μ²λ¦¬νκ³ μΈν¬μ μμ‘΄μ¨κ³Ό μΈν¬λ΄ νμ±μ°μμ’
μ λλ λ³νλ₯Ό μΈ‘μ νμλ€.
κ²°κ³Ό: μμ€μ½λ₯΄λΈμ°κ³Ό μμ€νμν΄μ μ²λ¦¬νμ λ κ³Όμ°νμμμ μ€νμ₯ μμΈμ μΌλ‘ μΈν μΈν¬μ μμμ΄ μ μνκ² κ°μνμλ€. 0.2mMμ 0.4mMμ κ³Όμ°νμμλ₯Ό μ²λ¦¬νμ λ μΈν¬λ κ°κ° 80%, 69%μ μμ‘΄μ¨μ 보μλ€. μ΄ μ‘°κ±΄μμ 500uMμ μμ€μ½λ₯΄λΈμ°μ μ²λ¦¬νμ λ κ°κ°μ 쑰건μμ μμ‘΄μ¨μ΄ 27% 14% μ¦κ°νλ€. 20mJ/cm2, 40mJ/cm2 μ μ€νμ₯μ μΈν¬μ μ²λ¦¬νμ λ 85%, 65%μ μμ‘΄μ¨μ 보μκ³ μ΄λ 500uM λλμ μμ€μ½λ₯΄λΈμ°μ μ²λ¦¬νμ λ κ°κ° 17%, 12% μ¦κ°νμλ€. κ° μ‘°κ±΄μμ μΈν¬ λ΄λΆμ νμ±μ°μμ’
μ λλκ° κ°μν κ²μ ν λλ‘ μΈν¬ μμ‘΄μ¨μ μ¦κ°λ μμ€μ½λ₯΄λΈμ°μ νμ°ν ν¨κ³Όμ κΈ°μΈν κ²μΌλ‘ νλ¨λλ€. μμ€νμν΄λ λ§μ°¬κ°μ§λ‘ νμ°ν ν¨κ³Όλ₯Ό λνλ΄μ΄ μΈν¬μ μμ‘΄μ¨μ μ¦κ°μμΌ°μ§λ§, μμ€μ½λ₯΄λΈμ°λ³΄λ€ κΈ΄ μ²λ¦¬ μκ°μ΄ νμνλ€. μΈν¬μ 0.2mMμ κ³Όμ°νμμλ‘ μ°νμ€νΈλ μ€λ₯Ό μ λ°νκ³ 90uMμ μμ€μ½λ₯΄λΈμ°κ³Ό 20uMμ μμ€νμν΄μ 30 μκ° λμ κ°μ΄ μ²λ¦¬νμ λ κ° μ½λ¬Όμ λ¨λ
μ²λ¦¬λ³΄λ€ ν₯μλ ν¨κ³Όλ₯Ό 보μ΄λ©° 75%μμ 129%λ‘ μΈν¬μ μμ‘΄μ¨μ μ¦κ°μμΌ°λ€.
κ²°λ‘ : κ³Όμ°νμμμ μ€νμ₯ μμΈμ μΌλ‘ μ λλ λ§λ§μμμνΌμΈν¬μ£Όμ μ°νμ€νΈλ μ€ λͺ¨λΈμ΄ νμ°ν ν보 λ¬Όμ§μ ν¨κ³Όλ₯Ό μ€ννλλ° νλΉν κ²μ λ°νλ€. μμ€μ½λ₯΄λΈμ°κ³Ό μμ€νμν΄μ΄ μ΄ λͺ¨λΈμμ νμ°ν ν¨κ³Όλ₯Ό λνλκ³ μμ€νμν΄μ νμ°ν λ¬Όμ§λ‘ μμ©νλλ° μμ©μ± νκ²½μμ μμ€μ½λ₯΄λΈμ°λ³΄λ€ κΈ΄ μ²λ¦¬μκ°μ΄ νμν κ²μ λ°νκ³ μμ€μ½λ₯΄λΈμ°κ³Ό μμ€νμν΄μ νΌν©νμ¬ μ²λ¦¬νλ©΄ κ° μ½λ¬Όμ λ¨λ
μ²λ¦¬λ³΄λ€ λ°μ΄λ νμ°ν ν¨κ³Όλ₯Ό λνλ΄λ κ²μ νμΈνμλ€.1. Introduction 1
2. Materials and methods 4
2.1 ARPE-19 cell culture 4
2.2 Hydrogen peroxide exposure procedure 4
2.3 Ultraviolet B irradiation procedure 5
2.4 DPPH ROS scavenging assay 5
2.5 Antioxidant treatment 6
2.6 MTT assay 6
2.7 Crystal violet assay 7
2.8 DCFH-DA intracellular ROS level assay 7
3. Results 9
4. Discussion 28
5. References 32Maste
μ νμ λ³κ²½ νκΈ μ°¨λ¨μ μν μΈν°νμ΄μ€ μ¬μ€κ³ λ°©λ²λ‘
νμλ
Όλ¬Έ(μμ¬) --μμΈλνκ΅ λνμ :μ°μ
곡νκ³Ό,2008.2.Maste
Animal modelμ μν μ μμ κ²½μ νμ§μ μ μ λͺ¨μ μΆμ μ κ΄ν μ°κ΅¬
νμλ
Όλ¬Έ(μμ¬)--μμΈλνκ΅ λνμ :λλ¬Όμμκ³Όνκ³Ό κ°μΆμ‘μ’
νμ 곡,1997.Maste