Xing Z, Jordana M, Kirpalani H, Driscoll K E, Schall T J, Gauldie J
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
Am J Respir Cell Mol Biol. 1994 Feb;10(2):148-53. doi: 10.1165/ajrcmb.10.2.8110470.
Using a rat model of acute lung inflammation induced by intratracheal instillation of lipopolysaccharide (LPS), we investigated the kinetics of mRNA expression and the potential cellular sources of tumor necrosis factor-alpha (TNF-alpha), macrophage inflammatory protein-2 (MIP-2), interleukin (IL)-1 beta, IL-6, RANTES, and transforming growth factor-beta 1 (TGF-beta 1). By Northern blot analysis, TNF-alpha and MIP-2 mRNAs in total lung tissue increased markedly by 30 min and peaked by 1 h after LPS exposure, whereas expression of IL-1 beta and IL-6 was not detected until 1 h and peaked within 6 h. In contrast, neither RANTES nor TGF-beta 1 mRNA was induced by LPS throughout 72 h, although a basal expression was detected in both saline- and LPS-treated lung tissues. At 1 h after LPS, the bronchoalveolar lavage (BAL) fluid contained about 98% alveolar macrophages (AM), whereas by 6 or 12 h, 88% of BAL cells were polymorphonuclear neutrophils (PMN). Upon extraction of total RNA after separation of AM from PMN in BAL, Northern analysis showed that at 1 h, AM expressed pronounced signals for TNF-alpha, MIP-2, IL-1 beta, and IL-6. At 6 and 12 h, however, while cytokine transcripts decreased in AM, PMN exhibited strong signals for these cytokines. A low basal noninducible signal for TGF-beta 1 but not RANTES was detected in both AM and PMN. Finally, by in situ hybridization techniques, PMN in the lung tissue, particularly those located in the vicinity of the bronchiole and vasculature, were demonstrated to localize MIP-2 mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)
利用气管内注入脂多糖(LPS)诱导的大鼠急性肺炎症模型,我们研究了肿瘤坏死因子-α(TNF-α)、巨噬细胞炎性蛋白-2(MIP-2)、白细胞介素(IL)-1β、IL-6、调节激活正常T细胞表达和分泌因子(RANTES)以及转化生长因子-β1(TGF-β1)的mRNA表达动力学及其潜在细胞来源。通过Northern印迹分析,LPS暴露后30分钟,全肺组织中的TNF-α和MIP-2 mRNA显著增加,并在1小时达到峰值,而IL-1β和IL-6的表达直到1小时才被检测到,并在6小时内达到峰值。相比之下,尽管在盐水处理和LPS处理的肺组织中均检测到基础表达,但在整个72小时内,LPS均未诱导RANTES和TGF-β1 mRNA表达。LPS处理1小时后,支气管肺泡灌洗(BAL)液中约98%为肺泡巨噬细胞(AM),而在6或12小时时,88%的BAL细胞为多形核中性粒细胞(PMN)。从BAL中的AM和PMN分离后提取总RNA,Northern分析显示,在1小时时,AM表达了明显的TNF-α、MIP-2、IL-1β和IL-6信号。然而,在6和12小时时,虽然AM中的细胞因子转录本减少,但PMN表现出这些细胞因子的强信号。在AM和PMN中均检测到低水平的基础非诱导性TGF-β1信号,但未检测到RANTES信号。最后,通过原位杂交技术,肺组织中的PMN,特别是位于细支气管和脉管系统附近的PMN,被证明可定位MIP-2 mRNA。(摘要截短于250字)