MIGRATION AND PROLIFERATION OF GUINEA-PIG AND HUMAN AIRWAY EPITHELIAL-CELLS IN RESPONSE TO TACHYKININS
MIGRATION AND PROLIFERATION OF GUINEA-PIG AND HUMAN AIRWAY EPITHELIAL-CELLS IN RESPONSE TO TACHYKININS
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DOI：
10.1152/ajplung.1995.269.1.l119
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发表时间：
1995-07-01
影响因子：
4.9
通讯作者：
WHITE, SR
中科院分区：
文献类型：
--
作者：
KIM, JS;RABE, KF;WHITE, SR
受损气道上皮层的恢复是气道修复的必要组成部分。速激肽，包括 P 物质 (SP) 和神经激肽 A (NKA)，定位于气道粘膜内的感觉神经。这些速激肽调节多种气道功能，但它们在上皮修复中的作用尚未被探索。为了确定速激肽是否刺激气道上皮细胞的迁移和增殖，将豚鼠气管上皮（GPTE）和人支气管上皮（HBE）细胞在原代培养物中生长4-5天。在盲井趋化室中评估上皮细胞迁移，并在掺入胸苷类似物 5-溴-2'-脱氧尿苷 (BrdU) 后通过免疫组织化学测定增殖情况。 GPTE 和 HBE 细胞在用 10(-11) M NKA 刺激后均发生迁移 [每 10 个高倍视野 (hpf) 28.0 +/- 3.6 与 5.4 +/- 1.2 个细胞，P < 0.001，对于 GPTE 细胞，n = 8；对照为每 10 hpf 18.4 +/- 2.3 与 3.8 +/- 0.5 个细胞，P < 0.001，对于 HBE 细胞，n = 4]。迁移在 2 小时内受到刺激，6 小时后达到最大，并被神经激肽 2 (NKA) 受体拮抗剂 SR-48968 显着减弱。 NKA 刺激的迁移既具有趋化性，又具有趋化性，并且可以通过用环己酰胺抑制蛋白质合成、用秋水仙碱抑制微管功能或用细胞松弛素 D 抑制肌动蛋白微丝伸长来阻断。特定 NK1 受体激动剂 Sar(9)-P 物质 (SP) 也可以刺激迁移，但反应幅度小于 NKA。 Sar(9)-SP 可刺激 GPTE 细胞穿越 S 期，但 NKA 则不会。 10(-10) M Sar(9)-SP 处理使所有细胞的 BrdU 标记从 2.7 +/- 0.7 增加到 12.1 +/- 3.6%（P < 0.05，n = 5）。用 10(-10) M Sar(9)-SP 刺激 72 小时，细胞数量从 143,300 +/- 42,800 增加到 205,000 +/- 42,700（P < 0.05，It = 3）。我们证明NKA和Sar(9)-SP在原代培养物中引发GPTE细胞的迁移；然而，只有 Sar(9)-SP 能引起这些细胞的增殖。这些数据表明速激肽可以促进受损上皮的修复。
Restoration of the epithelial lining of a damaged airway is a necessary component of airway repair. Tachykinins, including substance P (SP) and neurokinin A (NKA), are localized to sensory nerves within the airway mucosa. These tachykinins regulate several airway functions, but their role in the repair of the epithelium has not been explored. To determine whether tachykinins stimulate migration and proliferation of airway epithelial cells, guinea pig tracheal epithelial (GPTE) and human bronchial epithelial (HBE) cells were grown in primary culture for 4-5 days. Epithelial cell migration was assessed in a blindwell chemotaxis chamber, and proliferation was determined by immunohistochemistry after incorporation of the thymidine analogue 5-bromo-2'-deoxyuridine (BrdU). Both GPTE and HBE cells migrated after stimulation with 10(-11) M NKA [28.0 +/- 3.6 vs. 5.4 +/- 1.2 cells per 10 high-power fields (hpf), P < 0.001, n = 8 for GPTE cells; 18.4 +/- 2.3 vs. 3.8 +/- 0.5 cells per 10 hpf for control, P < 0.001, n = 4 for HBE cells]. Migration was stimulated within 2 h, was maximal after 6 h, and was attenuated substantially by the neurokinin 2 (NKA)-receptor antagonist SR-48968. NKA-stimulated migration was both chemokinetic and chemotactic, and it could be blocked by inhibition of protein synthesis with cyclohexamide, inhibition of microtubular function with colchicine, or inhibition of actin microfilament elongation with cytochalasin D. Migration was also stimulated by the specific NK1-receptor agonist Sar(9)-substance P (SP), though the magnitude of response was less than for NKA. Traversal of S phase was stimulated in GPTE cells by Sar(9)-SP but not by NKA. Treatment with 10(-10) M Sar(9)-SP increased BrdU labeling from 2.7 +/- 0.7 to 12.1 +/- 3.6% of all cells (P < 0.05, n = 5). Stimulation with 10(-10) M Sar(9)-SP for 72 h increased cell numbers from 143,300 +/- 42,800 to 205,000 +/- 42,700 (P < 0.05, It = 3). We demonstrate that NKA and Sar(9)-SP elicit migration of GPTE cells in primary culture; however, only Sar(9)-SP elicits proliferation of these cells. These data suggest that tachykinins may facilitate repair of a damaged epithelium.