脂多糖对气道上皮细胞分泌炎症因子的影响

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。

The inflation of lipopolysaccharide to cytokine in airway epithelial cell

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 .