CAJ | 학술논문

目的：：本研究旨在揭示两种氧化物过氧化氢(H 2 O 2)及氧化低密度脂蛋白(ox-LDL)对视网膜色素上皮细胞(retinal pigmented epithelium cells,RPE)的氧化损伤作用,为进一步深入开展抗氧化应激保护治疗提供理论基础及依据。方法：体外培养人 ARPE-19细胞系,ox-LDL 及 H 2 O 2分别干预培养细胞诱导氧化应激损伤,采用 MTS 法及 AVFI/PI 联合流式细胞仪对 RPE 细胞增殖率、凋亡率进行分析测定；通过荧光标记对细胞内生活性氧(ROS)水平及 RPE 的细胞衰老水平进行半定量分析检测,从而了解及评估上述这两种氧化物对RPE 氧化应激损伤。结果：1)ox-LDL 及 H 2 O 2干预后,细胞内生活性氧族(ROS)水平显著上升,检测得到荧光强度平均值分别为对照组537.495,H 2 O 2干预组958.533,ox-LDL 干预组644.994,与正常对照组相比分别为P =0.040及 P =0.016,且两种氧化物相比,H 2 O 2组 ROS 水平比 ox-LDL 组上升更加显著；2)ox-LDL 及 H 2 O 2干预后,RPE 细胞增值率显著下降,与正常对照组相比分别为 P =0.020及 P =0.026；而细胞凋亡率两组均上升,与正常对照组相比分别为 P =0.013及 P =0.003；3)就细胞衰老状态而言,两者均使细胞衰老状态显著,检测得到荧光强度平均值分别为对照组218.571,H 2 O 2干预组320.240,ox-LDL 干预组293.868,与正常对照组相比 P =0.045,但组间无明显差异。结论：ox-LDL 及 H 2 O 2均能造成 RPE 氧化应激损伤,能造成细胞 ROS 含量显著增加,并导致凋亡及细胞衰老状态明显,为深入开展老年黄斑变性的研究奠定基础。
목적：：본연구지재게시량충양화물과양화경(H 2 O 2)급양화저밀도지단백(ox-LDL)대시망막색소상피세포(retinal pigmented epithelium cells,RPE)적양화손상작용,위진일보심입개전항양화응격보호치료제공이론기출급의거。방법：체외배양인 ARPE-19세포계,ox-LDL 급 H 2 O 2분별간예배양세포유도양화응격손상,채용 MTS 법급 AVFI/PI 연합류식세포의대 RPE 세포증식솔、조망솔진행분석측정；통과형광표기대세포내생활성양(ROS)수평급 RPE 적세포쇠로수평진행반정량분석검측,종이료해급평고상술저량충양화물대RPE 양화응격손상。결과：1)ox-LDL 급 H 2 O 2간예후,세포내생활성양족(ROS)수평현저상승,검측득도형광강도평균치분별위대조조537.495,H 2 O 2간예조958.533,ox-LDL 간예조644.994,여정상대조조상비분별위P =0.040급 P =0.016,차량충양화물상비,H 2 O 2조 ROS 수평비 ox-LDL 조상승경가현저；2)ox-LDL 급 H 2 O 2간예후,RPE 세포증치솔현저하강,여정상대조조상비분별위 P =0.020급 P =0.026；이세포조망솔량조균상승,여정상대조조상비분별위 P =0.013급 P =0.003；3)취세포쇠로상태이언,량자균사세포쇠로상태현저,검측득도형광강도평균치분별위대조조218.571,H 2 O 2간예조320.240,ox-LDL 간예조293.868,여정상대조조상비 P =0.045,단조간무명현차이。결론：ox-LDL 급 H 2 O 2균능조성 RPE 양화응격손상,능조성세포 ROS 함량현저증가,병도치조망급세포쇠로상태명현,위심입개전노년황반변성적연구전정기출。
Objective:To reveal the oxidative damage effect on retinal pigment epithelial cells of two oxides hydrogen peroxide (H 2 O 2 )and oxidized low density lipoprotein (ox-LDL),and provide theoretical basis and foundation for antioxidant stress protection treatment.Methods:In vitro cultured ARPE-1 9 cell line,ox-LDL and H 2 O 2 respectively intervented cultured cells induced oxidative stress injury,cell proliferation and apoptosis rate were detected with flow cy-tometry combined AVFI/PI and MTS;the reactive oxygen species (ROS)of RPE and RPE cell senescence were determined by semi quantitative analysis with fluorescent labeling.Results:1 )The ROS level increased significantly compared with the normal control group after the intervention by ox-LDL and H 2 O 2 ,average size of fluorescence intensity were 537.495 and 958.533 and 644.994,respectively P =0.040 and P =0.01 6;and the ROS levels between two kinds of oxides(H 2 O 2 and ox-LDL)had significant difference;2)cell proliferation of RPE after the intervention by ox-LDL and H 2 O 2 was reduced significantly,respectively P = 0.020 and P =0.026;apoptosis rate of RPE after the intervention by ox-LDL and H 2 O 2 was increased sig-nificantly,respectively P =0.013及 P =0.003;3)the cell senescent state of two kinds of oxides (H 2 O 2 and ox-LDL)were both increased significantly compared with the normal cells,average size of fluorescence intensity were 218.571,320.240 and 293.868,and P = 0.045,but had no significant difference between H 2 O 2 and ox-LDL groups.Conclusion:Ox-LDL and H 2 O 2 can both cause oxidative stress injury of RPE,and a dramatic increase in ROS levels,and lead to ap-optosis and cell senescence obviously,and lay the foundation for the further study of age-related macular degeneration (AMD).