CAJ | 학술논문

目的探讨地塞米松对氧化应激时H441细胞肺表面活性蛋白(surfactant protein,SP)A、B、C和D表达的影响. 方法采用H2O2制作H441细胞氧化应激模型.实验分为对照组、不同浓度地塞米松(1、10和100 μmol/L)组、H2O2(100 μmol/L)损伤组、H2O2(100 μmol/L)+地塞米松(100 μmol/L)组.共培养24 h后采用实时荧光定量-聚合酶链反应技术检测各组培养细胞中SP A、B、C和D mRNA的表达水平,采用Western印迹技术检测SP-A、B、C和D蛋白的表达.多组间比较采用单因素方差分析,并采用Dunneu's T3检验进行两两比较. 结果 (1)H2O2损伤组SPA mRNA表达水平明显高于对照组(4.0016±0.3618与0.9914±0.4170),而SP C与SP-D表达明显低于对照组(SP-C:0.2840±0.0586与0.9719±0.1022; SP-D:0.6199±0.0074与0.9518±0.0614),差异均有统计学意义(P均＜0.05);与H2O2损伤组相比,H 2O2+地塞米松组SP-AmRNA的表达水平下降(2.5191±0.3463),而SP-D mRNA的表达水平升高(0.9076±0.0167),差异均有统计学意义(P均＜0.05).(2)H2O2损伤组SP-A蛋白表达水平明显高于对照组(2.1795±0.0645与0.9728±0.0090),SP-B、SP-C与SP D蛋白表达水平明显低于对照组(SP-B:0.2593±0.0086与0.7253±0.0134; SP C:0.5592±0.0111与0.9472±0.0052;SP-D:1.1550±0.0237与1.2991±0.0298);与H2O2损伤组相比,H2O2+地塞米松组SP-A蛋白表达降低(1.4974±0.0185),而SP B、SP-C与SP D蛋白表达水平升高(0.7389±0.0182、0.9244±0.0282和1.3339±0.0136),差异均有统计学意义(P均＜0.05). 结论地塞米松可能通过调节SP的表达,对H441细胞氧化应激损伤发挥保护作用.
목적 탐토지새미송대양화응격시H441세포폐표면활성단백(surfactant protein,SP)A、B、C화D표체적영향. 방법 채용H2O2제작H441세포양화응격모형.실험분위대조조、불동농도지새미송(1、10화100 μmol/L)조、H2O2(100 μmol/L)손상조、H2O2(100 μmol/L)+지새미송(100 μmol/L)조.공배양24 h후채용실시형광정량-취합매련반응기술검측각조배양세포중SP A、B、C화D mRNA적표체수평,채용Western인적기술검측SP-A、B、C화D단백적표체.다조간비교채용단인소방차분석,병채용Dunneu's T3검험진행량량비교. 결과 (1)H2O2손상조SPA mRNA표체수평명현고우대조조(4.0016±0.3618여0.9914±0.4170),이SP C여SP-D표체명현저우대조조(SP-C:0.2840±0.0586여0.9719±0.1022; SP-D:0.6199±0.0074여0.9518±0.0614),차이균유통계학의의(P균＜0.05);여H2O2손상조상비,H 2O2+지새미송조SP-AmRNA적표체수평하강(2.5191±0.3463),이SP-D mRNA적표체수평승고(0.9076±0.0167),차이균유통계학의의(P균＜0.05).(2)H2O2손상조SP-A단백표체수평명현고우대조조(2.1795±0.0645여0.9728±0.0090),SP-B、SP-C여SP D단백표체수평명현저우대조조(SP-B:0.2593±0.0086여0.7253±0.0134; SP C:0.5592±0.0111여0.9472±0.0052;SP-D:1.1550±0.0237여1.2991±0.0298);여H2O2손상조상비,H2O2+지새미송조SP-A단백표체강저(1.4974±0.0185),이SP B、SP-C여SP D단백표체수평승고(0.7389±0.0182、0.9244±0.0282화1.3339±0.0136),차이균유통계학의의(P균＜0.05). 결론 지새미송가능통과조절SP적표체,대H441세포양화응격손상발휘보호작용.
Objective To explore the effects of dexamethasone on the expressions of surfactant protein(SP) A,B,C and D in H441 cells injured by oxidative stress.Methods H441 cells were challenged with hydrogen peroxide as oxidative stress damage cell model.Then the cells were divided into control group,groups on different dexamethasone levels (1,10 and 100 umol/L),H2O2 (100 μmol/L) injury group and dexamethasone (100 μmol/L)+ H2 O2 (100 μmol/1) group.All cells were cultured for 24 hours,then the expressions of SP mRNA were analyzed by quantitative real timepolymerase chain reaction,and proteins were detected by Western blot.One-way variant analysis and Dunneu's T3 test were applied as statistical methods.Results (1) The mRNA level of SP-A in H2 O2 injury group was higher than that in the control group (4.0016±0.3618 vs 0.9914±0.4170),while mRNA of SP C and SP-D were lower than those in the control group (SP C:0.2840±0.0586 vs 0.9719±0.1022;SP-D:0.6199±0.0074 vs0.9518±0.0614) (all P＜0.05).Compared with H2O2injury group,the mRNA level of SP-A in dexamethasone + H2O2 group was down-regulated (2.5191±0.3463),but SP-D mRNA was up-regulated (0.9076±0.0167)(all P＜0.05).(2) The SP-A protein level in H2O2 injury group was higher than that in the control group (2.1795±0.0645 vs 0.9728±0.009),while SP-B、SP-C and SP-D protein levels were lower than those in the control group (SP-B:0.2593±0.0086 vs 0.7253±0.0134;SP-C:0.5592±0.0111 vs 0.9472±0.0052;SP-D:1.1550±0.0237 vs 1.2991±0.0298).Compared with H2O2 injury group,the protein level of SP-A in dexamethasone+ H2 O2 group was down-regulated(1.4974 ± 0.0185),while SP-B,SP-C and SP-D levels were up-regulated (0.7389±0.0182,0.9244±0.0282 and 1.3339±0.0136).The differences were statistically significant respectively (all P＜0.05).Conclusions Dexamethasone may protect H441 cells from being damaged by H2O2 through regulating SP mRNA and protein expression.