CAJ | 학술논문

目的：检测不同浓度脂多糖（LPS）刺激气道上皮细胞不同时间后炎症因子[转化生长因子（TGF）‐β、单核细胞趋化蛋白‐1（MCP‐1）、肿瘤坏死因子α（TNF‐α）、细胞间黏附分子（ICAM）‐1、ICAM‐2]分泌量的变化，从而探索LPS在调节气道上皮细胞炎症因子分泌中的作用。方法用培养贴壁细胞的方法，培养具有气道上皮细胞特性的A549细胞株，用不同浓度的LPS（0、0．1、1．0、10．0、20．0、30．0、50．0、100．0μg／mL ）刺激饥饿24 h后的同等数量的A549细胞，分别在刺激2、4、8、24、30 h以后收集细胞上清液；用ELASA方法检测A549细胞各炎症因子分泌量，分析不同时间点的变化。结果（1）LPS能够干预气道上皮细胞分泌TGF‐β、MCP‐1、TNF‐α、ICAM‐1和ICAM‐2；当LPS干预浓度为20．0μg／mL ，干预时间为4 h和8 h时，ICAM‐1的分泌量达到峰值；当LPS干预浓度为50．0μg／mL ，干预时间为4 h时，MCP‐1的分泌量达到峰值。（2）TGF‐β、MCP‐1、TNF‐α、ICAM‐1和ICAM‐2的分泌量与LPS的干预浓度和干预时间有一定关系，在各个时相点，TNF‐α、MCP‐1和ICAM‐1分泌量达到最大时与空白对照组相比，差异有统计学意义（P＜0．01）；在干预时间为30 h时，TGF‐β分泌量下降，ICAM‐2的分泌量与空白对照组相比，差异无统计学意义（P＞0．05）；在其余各时相点，TGF‐β和ICAM‐2分泌量最大时也需要一个最佳的LPS干预浓度，且与空白对照组比较，差异有统计学意义（P＜0．01）。结论 LPS对气道上皮细胞TGF‐β、MCP‐1、TNF‐α、ICAM‐1和ICAM‐2分泌量的影响与L PS的干预浓度和干预时间相关，特定的干预浓度和干预时间可使相应的炎症因子分泌达峰值。
목적：검측불동농도지다당（LPS）자격기도상피세포불동시간후염증인자[전화생장인자（TGF）‐β、단핵세포추화단백‐1（MCP‐1）、종류배사인자α（TNF‐α）、세포간점부분자（ICAM）‐1、ICAM‐2]분비량적변화，종이탐색LPS재조절기도상피세포염증인자분비중적작용。방법용배양첩벽세포적방법，배양구유기도상피세포특성적A549세포주，용불동농도적LPS（0、0．1、1．0、10．0、20．0、30．0、50．0、100．0μg／mL ）자격기아24 h후적동등수량적A549세포，분별재자격2、4、8、24、30 h이후수집세포상청액；용ELASA방법검측A549세포각염증인자분비량，분석불동시간점적변화。결과（1）LPS능구간예기도상피세포분비TGF‐β、MCP‐1、TNF‐α、ICAM‐1화ICAM‐2；당LPS간예농도위20．0μg／mL ，간예시간위4 h화8 h시，ICAM‐1적분비량체도봉치；당LPS간예농도위50．0μg／mL ，간예시간위4 h시，MCP‐1적분비량체도봉치。（2）TGF‐β、MCP‐1、TNF‐α、ICAM‐1화ICAM‐2적분비량여LPS적간예농도화간예시간유일정관계，재각개시상점，TNF‐α、MCP‐1화ICAM‐1분비량체도최대시여공백대조조상비，차이유통계학의의（P＜0．01）；재간예시간위30 h시，TGF‐β분비량하강，ICAM‐2적분비량여공백대조조상비，차이무통계학의의（P＞0．05）；재기여각시상점，TGF‐β화ICAM‐2분비량최대시야수요일개최가적LPS간예농도，차여공백대조조비교，차이유통계학의의（P＜0．01）。결론 LPS대기도상피세포TGF‐β、MCP‐1、TNF‐α、ICAM‐1화ICAM‐2분비량적영향여L PS적간예농도화간예시간상관，특정적간예농도화간예시간가사상응적염증인자분비체봉치。
Objective To detect different cytokines (TGF‐β,MCP‐1 ,TNF‐α,ICAM‐1 ,ICAM‐2) concentration after lipopo‐lysaccharides(LPS) stimulation airway epithelial cells at the same time ,to explore the role of LPS in the airway epithelial cells se‐creting a variety of cytokines .Methods A549 cell strain was cultivated and interfered by different concentrations LPS(0 ,0 .1 ,1 .0 , 10 .0 ,20 .0 ,30 .0 ,50 .0 ,100)μg/mL after 2 ,4 ,8 ,24 ,30 h .Collected culture medium and total protein in the cell ,detected volume of cytokines TGF‐β,MCP‐1 ,TNF‐α,ICAM‐1 and ICAM‐2 secreted by A549 cell using enzyme‐linked immunosorbent assays (ELISA) ,to analysed the changes of above‐mentioned cytokines at the different time period .Results First ,LPS interfered the air‐way epithelial cells secreting TGF‐β,MCP‐1 ,TNF‐α,ICAM‐1 and ICAM‐2 ;the secretion of ICAM‐1 peaked when the concentration of LPS was 20 .0μg/mL and the time was 4 h and 8 h;the secretion of MCP‐1 peaked when the concentration of LPS was 50 .0μg/mL and the time was 4 h .Second ,the volume of cytokines TGF‐β,MCP‐1 ,TNF‐α,ICAM‐1 ,ICAM‐2 was related to the concentra‐tion and the period of LPS .Compared with the control group ,the maximum secretion of TNF‐α,MCP‐1 and ICAM‐1 were increased at all the different time ,the difference was statistically significant (P<0 .01);whereas at the 30 h ,the secretion of TGF‐βwas de‐creased and the secretion of ICAM‐2 was not different from the control group(P> 0 .05) ,compared with the control group ,the maximum secretion of TGF‐βand ICAM‐2 were increased at all the other different time ,the difference was statistically significant (P<0 .01) .Conclusion LPS influenced the volume of cytokines TGF‐β,MCP‐1 ,TNF‐α,ICAM‐1 and ICAM‐2 are related to the time and concentration of LPS ,the particular intervention time and concentration enabled them reaches maximum .