CAJ | 학술논문

目的探讨氢气对中波紫外线(UVB)致人成纤维细胞氧化损伤的影响.方法原代培养人成纤维细胞,分组如下:正常对照组,未经任何处理；氢气对照组,富氢培养液处理；UVB照射组；富氢培养液后处理组；富氢培养液预处理组.通过噻唑蓝(MTT)法观察细胞增殖情况,通过化学发光和酶联免疫吸附试验(ELISA)观察超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性,丙二醛(MDA)和8异前列腺素2α(8-isoPGE2α)的水平,Western印迹观察血红素加氧酶-1(HO-1)蛋白的表达情况,采用单因素方差分析进行统计学处理.结果给予不同剂量的UVB照射,细胞增殖活性(A490)呈剂量依赖性降低.氢气后处理和预处理组细胞增殖活性明显高于相应UVB组(均P＜0.05).氢气后、预处理组SOD和CAT活性以及HO-1蛋白表达水平明显高于UVB组(均P＜0.05),但MDA和8-iso-PGE2α明显低于UVB组(均P＜0.05).结论氢气可以减轻UVB诱发的成纤维细胞氧化损伤.
목적 탐토경기대중파자외선(UVB)치인성섬유세포양화손상적영향.방법 원대배양인성섬유세포,분조여하:정상대조조,미경임하처리；경기대조조,부경배양액처리；UVB조사조；부경배양액후처리조；부경배양액예처리조.통과새서람(MTT)법관찰세포증식정황,통과화학발광화매련면역흡부시험(ELISA)관찰초양화물기화매(SOD)、과양화경매(CAT)적활성,병이철(MDA)화8이전렬선소2α(8-isoPGE2α)적수평,Western인적관찰혈홍소가양매-1(HO-1)단백적표체정황,채용단인소방차분석진행통계학처리.결과 급여불동제량적UVB조사,세포증식활성(A490)정제량의뢰성강저.경기후처리화예처리조세포증식활성명현고우상응UVB조(균P＜0.05).경기후、예처리조SOD화CAT활성이급HO-1단백표체수평명현고우UVB조(균P＜0.05),단MDA화8-iso-PGE2α명현저우UVB조(균P＜0.05).결론 경기가이감경UVB유발적성섬유세포양화손상.
Objective To observe the effect of hydrogen on ultraviolet B (UVB)-induced oxidative damage to skin fibroblasts.Methods Primary human skin fibroblasts from foreskin tissues were divided into five groups:normal control group receiving no treatment,hydrogen control group treated with hydrogen-rich saline,UVB group receiving irradiation only,post-treatment group irradiated with UVB followed by hydrogen-rich saline treatment,and pre-treatment group treated with hydrogen-rich saline followed by UVB irradiation.The dose of UVB was 30,60 and 90 mJ/cm2 in the cell proliferation assay and 90 mJ/cm2 in the other experiments.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of fibroblasts,a chemiluminescence method to estimate the activity of superoxide dismutase (SOD) and catalase as well as to determine the level of malondialdehyde in the culture supernatant of fibroblasts,enzyme linked immunosorbent assay (ELISA) to determine the supernatant level of 8-isoprostane-prostaglandin F2α (8-iso-PGF2α),Western blot to detect the expression of heme oxygenase-1 (HO-1) in fibroblasts.One-factor analysis of variance was conducted to assess differences in these parameters among these groups.Results UVB irradiation decreased the proliferative activity (absorbence value at 490 nm) of fibroblasts in a dose-dependent manner.Both the pre-treatment group and post-treatment group showed a statistical increase in proliferative activity of cells compared with the corresponding UVB control groups (all P ＜ 0.05).The activity of SOD and catalase as well as the protein expression of HO-1 were significantly higher (all P ＜ 0.05),whereas the supernatant levels of malondialdehyde and 8-iso-PGF2α were statistically lower (both P ＜ 0.05) in the pre-treatment group and post-treatment group than in the UVB control group.Conclusion Hydrogen may mitigate UVB-induced oxidative damage to skin fibroblasts.