Koyama S, Sato E, Masubuchi T, Takamizawa A, Nomura H, Kubo K, Nagai S, Izumi T
Shinshu University School of Medicine, First Department of Internal Medicine, Matsumoto 390, Japan.
Am J Physiol. 1998 Aug;275(2):L223-30. doi: 10.1152/ajplung.1998.275.2.L223.
We determined whether human lung fibroblasts (HLFs) might release mediators that are responsible for monocyte chemokinetic activity (MCA) constitutively. HLF supernatant fluids showed MCA in a time-dependent manner (P < 0.001). Checkerboard analysis of 24- and 72-h supernatant fluids showed that the activity was chemokinetic. Partial characterization of 24- and 72-h supernatant fluids revealed that the mediators released after 24 h were predominantly composed of lipid-soluble activity, and MCA was blocked by lipoxygenase inhibitors. The mediators released after 72 h were predominantly trypsin sensitive and blocked by cycloheximide. Molecular-sieve column chromatography identified four peaks of MCA. A polyclonal antibody to monocyte chemoattractant protein-1 (MCP-1) inhibited MCA by 20% after 24 h and by 40% after 72 h. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and transforming growth factor-beta (TGF-beta) antibodies attenuated MCA released after 72 h by 30 and 10%, respectively. These antibodies inhibited corresponding molecular-weight peaks separated by molecular-sieve column. The concentrations of MCP-1, GM-CSF, and TGF-beta were 4,698 +/- 242, 26.8 +/- 3.8, and 550 +/- 15 pg/ml, respectively. A leukotriene B4 (LTB4)-receptor antagonist attenuated the total MCA and the lowest molecular weight peak of MCA. The concentrations of LTB4 were 153.4 +/- 12.4 (24 h) and 212 +/- 16.6 (72 h) pg/ml. These findings suggest that HLFs may modulate the recruitment of monocytes into the lung by releasing MCP-1, GM-CSF, TGF-beta, and LTB4 constitutively.
我们确定了人肺成纤维细胞（HLFs）是否可能组成性地释放负责单核细胞趋化活性（MCA）的介质。HLF 上清液以时间依赖性方式显示出 MCA（P < 0.001）。对 24 小时和 72 小时上清液的棋盘分析表明该活性是趋化性的。对 24 小时和 72 小时上清液的部分特性分析显示，24 小时后释放的介质主要由脂溶性活性组成，且 MCA 被脂氧合酶抑制剂阻断。72 小时后释放的介质主要对胰蛋白酶敏感并被放线菌酮阻断。分子筛柱色谱法鉴定出四个 MCA 峰。抗单核细胞趋化蛋白-1（MCP-1）的多克隆抗体在 24 小时后使 MCA 降低 20%，在 72 小时后降低 40%。粒细胞-巨噬细胞集落刺激因子（GM-CSF）和转化生长因子-β（TGF-β）抗体分别使 72 小时后释放的 MCA 降低 30%和 10%。这些抗体抑制了通过分子筛柱分离的相应分子量峰。MCP-1、GM-CSF 和 TGF-β的浓度分别为 4,698±242、26.8±3.8 和 550±15 pg/ml。白三烯 B4（LTB4）受体拮抗剂减弱了总 MCA 和 MCA 的最低分子量峰。LTB4 的浓度分别为 153.4±12.4（24 小时）和 212±16.6（72 小时）pg/ml。这些发现表明 HLFs 可能通过组成性地释放 MCP-1、GM-CSF、TGF-β和 LTB4 来调节单核细胞向肺内的募集。
