论文部分内容阅读
目的:高氧治疗是某些新生儿疾病中不可或缺的治疗措施，长期治疗性高氧可能对肠上皮细胞产生严重的损伤作用，该研究探讨高氧是否通过活性氧(reactive oxygen species，ROS)促进肠上皮细胞凋亡信号调节激酶1(apoptosis signal-regulating kinase 1，ASK1)和P38的表达。方法:体外用不同浓度的过氧化氢(hydrogen peroxide，Hn 2On 2)(100 μmol/L、200 μmol/L和400 μmol/L)和85%氧气分别处理人结肠癌细胞系Caco-2细胞，免疫荧光检测ASK1表达，Western Blot和Real-time PCR方法检测P38和p-P38表达。n 结果:随着Hn 2On 2浓度的增加ASK1的荧光强度逐渐增强，高氧组ASK1荧光强度明显强于对照组和Hn 2On 2处理组。随着Hn 2On 2浓度(100 μmol/L、200 μmol/L、400 μmol/L)的增加，P38蛋白相对表达量(0.21±0.02、0.28±0.13、0.44±0.07)和p-P38蛋白相对表达量(0.09±0.02、0.19±0.03、0.37±0.07)逐渐升高，200 μmol/L和400 μmol/L Hn 2On 2组P38 mRNA(4.03±0.68、3.94±0.71)表达明显高于100 μmol/L Hn 2On 2组(3.05±0.47)(n P<0.01)。高氧组P38蛋白、p-P38蛋白以及P38 mRNA相对表达量明显高于Hn 2On 2组(n P<0.01)。与对照组相比，高氧组和Hn 2On 2组P38、p-P38蛋白以及P38 mRNA明显升高(n P<0.01)。n 结论:高氧和Hn 2On 2干预下，肠上皮细胞ASK1、P38和p-P38的表达均增加，表明ROS参与高氧诱导ASK1的活化进而激活下游P38，对肠上皮细胞产生损伤。n “,”Objective:Hyperoxia is a necessary therapy in some neonatal diseases, and long-term therapeutic hyperoxia may induce severe damaging effects on intestinal epithelial cells.The aim of this study was to investigate whether hyperoxia could promote the expression of ASK1 and P38 in intestinal epithelial cells through ROS.Methods:The human colon adenocarcinoma cell line Caco-2 cells were treated with different concentrations of Hn 2On 2(100 μmol/L, 200 μmol/L and 400 μmol/L)and 85% oxygen in vitro.The expression of ASK1 was detected by immunofluorescence, and the expression of P38 and p-P38 were detected by Western Blot and Real-time PCR.n Results:With the increase of Hn 2On 2 concentration, the fluorescence intensity of ASK1 increased.The fluorescence intensity of ASK1 in the hyperoxia group was significantly stronger than that of the control group and the Hn 2On 2 groups.With the increase of Hn 2On 2 concentration(100 μmol/L、200 μmol/L、400 μmol/L), the expression of P38 protein(0.21±0.02, 0.28±0.13, 0.44±0.07)and p-P38 protein(0.09±0.02, 0.19±0.03, 0.37±0.07)gradually increased.The expression of P38 mRNA in 200 μmol/L and 400 μmol/L H n 2On 2 groups(4.03±0.68、3.94±0.71)was significantly higher than that in 100 μmol/L H n 2On 2 group(3.05±0.47)(n P<0.01). The expressions of P38 protein, p-P38 protein and P38 mRNA in the hyperoxia group were significantly higher than those in the Hn 2On 2 group(n P<0.01). Compared with the control group, the expressions of P38 protein, p-P38 protein and p38 mRNA in the hyperoxia group and Hn 2On 2 groups increased significantly(n P<0.01).n Conclusion:The expression of ASK1 and P38 in intestinal epithelial cells increased significantly under hyperoxia, which indicated that hyperoxia might activate ASK1 and thereby regulate the expression of downstream P38 through ROS, resulting in intestinal epithelial cells damage.