摘要
目的 观察不同浓度氧气对脂多糖(LPS)诱导的大鼠肺泡Ⅱ型上皮细胞(AT-Ⅱ)分泌炎性因子的影响并探讨其机制.方法 原代培养的AT-Ⅱ随机分为5组(n=6):A、B、C、D组,E组分别置于无菌培养箱:A箱37℃、21%O2、5% CO2,B箱37℃、21%O2、5% CO2,C箱37℃、40%O2、55%N2、5% CO2,D箱37℃、60%O2、35%N2、5% CO2,E箱37℃、90%O2、5% N2、5%CO2;24 h后酶联免疫吸附试验(ELISA)检测各组细胞白细胞介素-8(IL-8)含量,反转录-聚合酶链反应(PT-PCR)及Western blot检测各组细胞p38丝裂原活化蛋白激酶(p38MAPK) mRNA及蛋白表达水平.结果 A、B、C、D、E组IL-8含量分别为(40.22 ±7.25)、(109.35±10.19)、(213.98±30.51)、(369.71 ±21.73)、(489.32 ±42.35) ng/L;p38MAPK mRNA表达水平分别为0.066± 0.010、0.163±0.008、0.228 ±0.027、0.268 ±0.016、0.292±0.010;p38 MAPK蛋白表达水平分别为0.055±0.005、0.124 ±0.006、0.182±0.009、0.227±0.013、0.303±0.010.与A组比较B、C、D、E组IL-8含量、p38MAPK mRNA和蛋白表达水平明显升高,差异有统计学意义(P<0.05);与B组比较C、D、E组IL-8含量、p38MAPK mRNA和蛋白表达水平逐渐升高,差异有统计学意义(P<0.05);C、D、E组间两两比较IL-8含量、p38MAPK mRNA和蛋白表达水平逐渐升高,差异有统计学意义(P<0.05).结论 40%~90%的氧气可使LPS诱导的AT-Ⅱ分泌IL-8呈浓度依赖性增加,其机制可能与胞内p38MAPK表达上调有关.
Objective To investigate the secretion of inflammatory cytokines in lipopolysaccharide (LPS)-induced aveolar epithelium type Ⅱ cells (AT-Ⅱ) cells exposed to various oxygen concentrations,and the possible mechanism.Methods Primary cultured AT-Ⅱ cells were randomly divided into five groups:A,B,C,D,and E,and then placed in sterilized incubators:37 ℃,21% O2,5% CO2 ;37 ℃,21% O2,5% CO2; 37 ℃,40% O2,55% N2,5% CO2; 37 ℃,60% O2,35% N2,5% CO2;37 ℃,90% O2,5% N2,5% CO2.Towenty-four h later,the supernatant fluid was used to assay the content of interleukin-8 (IL-8) by enzyme linked immunosorbent assay (ELISA).The expression of p38 mitogen-activated protein kinase (p38MAPK) mRNA and protein was detected in AT-Ⅱ cells by reverse transcription-polymerase chain reaction and Western blotting.Results ELISA,reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting showed that the content of IL-8 was (40.22±7.25),(109.35 ±10.19),(213.98±30.51),(369.71 ±21.73) and (489.32± 42.35) ng/L,p38MAPK mRNA expression level was 0.066 ± 0.010,0.163 ± 0.008,0.228 ± 0.027,0.268 ±0.016 and 0.292 ±0.010,and p38MAPK protein expression level was 0.055 ±0.005,0.124 ± 0.006,0.182 ±0.009,0.227 ±0.013 and 0.303 ±0.010 in groups A,B,C,D and E respectively.As compared with group A,the content of IL-8 in the supernatant fluid,and the expression of p38MAPK mRNA and protein were significantly increased in groups B,C,D and E (P 〈 0.05).As compared with groups B,the content of IL-8 in the supernatant fluid,and the p38MAPK mRNA and protein expression level in groups C,D and E were gradually increased with the difference being statistically significant (P 〈 0.05).In different oxygen concentration groups,with the increases in oxygen concentrations,the content of IL-8 in the supematant fluid,and the expression of p38MAPK mRNA and protein expression level were gradually increased with the difference being statistically significant (P 〈 0.05).Condusion 40%-90% oxygen could concentrasion-dependently increase the LPS-induced secretion of IL-8 in AT-Ⅱ cells probably by up-regulating the intracellular p38MAPK expression.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第2期364-366,共3页
Chinese Journal of Experimental Surgery
基金
河南省科技厅基础与前言研究项目(122300410055)
关键词
高氧性肺损伤
肺泡Ⅱ型上皮细胞
脂多糖
白细胞介素-8
p38丝裂原活
化蛋白激酶
Hyperoxia -induced lung injury
Aveolar epithelium type Ⅱ cells
Lipopolysaccharide
Interleukin - 8
p38 mitogen - activated protein kinase