摘要
目的观察机械牵张诱导肺泡巨噬细胞（AM）的细胞因子表达谱，以及机械牵张对脂多糖（LPS）诱导巨噬细胞炎性蛋白2（MIP-2）表达的影响。方法用LiquiChip液相蛋白芯片系统检测机械牵张诱导AM15种细胞因子的水平变化；检测不同强度机械牵张（5％、10％、15％和20％）和牵张刺激（20％）后不同时间点（0、1、3、6、12和24h）AM分泌MIP-2的水平，以及机械牵张（20％）与LPS（10ng／mL）共同刺激对AM分泌MIP-2的影响。结果牵张刺激后AM分泌IL-1β、IL-6、MIP-2、单核细胞趋化蛋白1（MCP-1）、IFN-γ和干扰素诱导蛋白10（IP-10）的水平明显升高（P均〈0．001）。机械牵张以强度和时间依赖方式诱导MIP-2的分泌，随着牵张强度的增加（10％、15％和20％），MIP-2的分泌水平也明显增加（P均〈0．001）；在刺激后6～24h范围内MIP-2水平持续升高（P〈0．001）。以机械牵张和LPS共同刺激AM，MIP-2的生成量大大增加，二者存在协同效应（F=121．983，P〈0．001）。结论机械牵张可诱导AM释放多种细胞因子，机械牵张诱导MIP-2的上调具有强度和时间依赖性，并协同LPS诱导AM释放MIP-2，可能在呼吸机所致肺损伤的发生和发展中起重要作用。
Objective To observe the effects of mechanical stretch on cytokines release from alveolar macrophages（AMs） and the expression of macrophage inflammatory protein-2 （MIP-2） induced by lipopolysaccharide（LPS）. Methods AMs were divided into the following groups：（1）AMs were subjected to 20% elongation by Flexercell 4000T cell stress system for 24 hours and the supernatant was collected to detect the levels of TNF-α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12,IFN-γ, macrophage inflammatory protein-1α （MIP-1α）, MIP-2, monocyte chemoattractant protein-1 （MCP-1）, granulocyte/maerophage colony stimulating factors （GM-CSF） , interferon inducible protein-10 （IP-10） , regulated on activation in normal T-cell expressed and secreted （ Rantes ） and keratinocyte chemoattractant （ KC ） , by using LiquiChip system. （2） AMs were subjected to 5% , 10% , 15% and 20% elongation for 24 hours and the supernatant was collected to detect the levels of MIP-2. （3）AMs were subjected to 20% elongation and MIP-2 in supernatant was detected 1,3, 6,12, and 24 hours later. （4） AMs were subjected to 20% elongation and/or LPS at a concentration of 10 ng/mL,and MIP-2 in supernatant was detected 24 hours later. Unstretehed AMs were used as control in all kind of test. Results （1）The levels of IL-113,IL-6, MIP-2,MCP-1, IFN-γ and IP-10 secreted by stretched AMs were 8.7,4. 3,38.6,4. 8,14.2 and 5.0 times those of the control group（ all P 〈0. 001 ）. （2） The levels of MIP-2 secreted by AMs subjected to 10% , 15% and 20% elongation were （480. 5 ± 93.1 ） pg/mL, （806. 3 ±225.9） pg/mL and （1335.7 ± 18.5） pg/mL respectively,all significantly higher than those of the control group [ （34.6 ± 11.4） pg,/mL,all P 〈0. 001 ]. （3） Three hours after the stimulation of stretch the level of MIP-2 began to increase gradually. And 6,12, and 24 hours after the stimulation the levels of MIP-2 secreted by the AMs were （819.4 ± 147.5） pg/mL, （1287.6 ± 380 ± 3） pg/mL and （
出处
《中国呼吸与危重监护杂志》
CAS
2009年第2期166-170,共5页
Chinese Journal of Respiratory and Critical Care Medicine
基金
广州市医药卫生科技项目(编号:2008-YB-012)
重点项目(编号:2008-ZDi-14)
