摘要
目的 观察大鼠肺泡巨噬细胞对内毒素 (LPS)重复刺激的耐受性与其Toll样受体 4(TLR4)表达的变化 ,研究两者之间的联系。方法 将 30只Wistar雄性大鼠分离所得肺泡巨噬细胞 ,用随机数字表法分为正常对照组 (A组 ) ,LPS单次刺激组 (B组 )和LPS 2次重复刺激组 (C组 )。用酶联免疫吸附测定 (ELISA)法和逆转录 聚合酶链反应 (RT PCR)法分别检测各组大鼠肺泡巨噬细胞分泌肿瘤坏死因子α(TNF α)及TLR4、白细胞介素 1 0 (IL 1 0 )、IL 1 8mRNA表达的变化 ,WesternBlot检测TLR4蛋白表达的变化。结果 大鼠肺泡巨噬细胞TNF α分泌和TLR4、IL 1 0、IL 1 8mRNA表达及TLR4的蛋白表达水平 ,A组分别为 (0 4 5 0± 0 0 1 0 ) μg/L、1 1 6± 0 0 4、0 97± 0 0 3、1 32 0± 0 0 2 0、5 8 1± 0 4 ;B组分别为 (0 76 0± 0 0 30 ) μg/L、2 1 8± 0 0 9、1 83± 0 0 7、2 0 6 0± 0 0 6 0、1 4 8.3± 1 4 ;B组与A组比较差异有显著性 (P <0 0 1 ) ;C组分别为 (0 4 90± 0 0 5 0 ) μg/L、1 2 3± 0 0 3、1 1 5± 0 0 5、1 1 70± 0 0 4 0、96 5±0 7;C组与B组比较差异也有显著性 (P <0 0 5或 <0 0 1 )。结论 LPS重复刺激可使大鼠肺泡巨噬细胞对LPS产生耐受性 ;
Objective To explore the change of expression of Toll like receptor 4 (TLR 4) related to the tolerance to lipopolysaccharide (LPS) in rat alveolar macrophages Methods Alveolar macrophages obtained from 30 Wistar male rats were randomly divided into a control group, a LPS single dose stimulation group and a LPS double dose stimulation group IL 10 and IL 18 mRNA in alveolar macrophages and the level of TNF α, as indexes of LPS tolerance, were measured by semi quantitative reverse transcription polymerase chain reaction (RT PCR) and enzyme linked immunoads orbeen assay (ELISA), respectively The expression of TLR 4 was detected by Western Blot Results The level of TNF α,and the mRNA for IL 10 and IL 18, and the protein of TLR 4 in the alveolar macrophages stimulated with single dose LPS (100 ng/ml) were highly increased as compared with the control group [(0 760±0 030) μg/L ,2 18±0 09, 1 83± 0 07, 2 060±0 060, 148 3±1 4 vs (0 450±0 010) μg/L,1 16±0 04, 0 97±0 03, 1 320± 0 020, 58 1±0 4( P <0 01)],while they were significantly decreased in the alveolar macrophages after re stimulation with double dose LPS (100 ng/ml and 500 ng/ml) Conclusions (1)The tolerance to LPS in alveolar macrophages can be induced by re stimulation with LPS (2) The down regulated expression of TLR 4 is related to tolerance to LPS
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2003年第7期411-414,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
国家自然科学基金资助项目 ( 3 973 0 2 10 )
