Calcineurin inhibitor effects on growth and phenotype of human airway epithelial cells In vitro
Calcineurin inhibitor effects on growth and phenotype of human airway epithelial cells In vitro
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DOI：
10.1111/j.1600-6143.2005.01071.x
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发表时间：
2005-11-01
影响因子：
8.8
通讯作者：
Randell, SH
中科院分区：
文献类型：
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作者：
Neuringer, IP;Sloan, J;Randell, SH
钙调磷酸酶抑制剂（CIs）环孢素和他克莫司构成肺移植免疫抑制的基础，但也对非淋巴组织产生生物学效应。随着吸入环孢素的出现，我们假设气道上皮也会受到高剂量环孢素的影响。本研究的目的是鉴定人气管支气管上皮细胞（hTBEC）钙调磷酸酶基因表达，并量化CIs对hTBEC生长、白细胞介素-1- β刺激的IL-8产生和hTBEC表型的影响。RT-PCR检测hTBEC培养物中Cyclophillin B和fk相关结合蛋白、calcalineurin A （α和β）、NFATC3和NFAT5。急性和慢性环孢素治疗1000 ng/mL显著抑制hTBEC增殖，而他克莫司则没有（急性治疗范围为10 ng/mL至1000 ng/mL，慢性治疗范围为50 ng/mL）。环孢素在10 000 ng/mL时显著增加分化良好的hTBEC培养物的LDH释放（n = 6），在1000 ng/mL时趋于显著。在环孢素处理的快速生长hTBEC培养物（n = 8）中，il - 1- β刺激IL-8的产生显著增加（p = 0.049）。长时间使用环孢素1000 ng/mL在气液界面（ALI）处理分化良好的hTBECs，可显著减少完整的多层粘膜纤毛上皮（p = 0.009）。对hTBEC生长的抑制、对IL-8产生的刺激以及对粘膜纤毛表型和完整多层上皮的长期影响提示环孢素可能对移植后气道上皮具有直接毒性作用。
Calcineurin inhibitors (CIs) cyclosporin and tacrolimus form the basis for immunosuppression in lung transplantation, yet also exert biological effects on nonlymphoid tissue. With the advent of inhaled cyclosporin, we hypothesize that the airway epithelium is also subject to CI effects at high doses. The aim of this study was to identify human tracheobronchial epithelial cell (hTBEC) calcineurin gene expression and quantify effects of CIs on hTBEC growth, interleukin-1-beta stimulated IL-8 production and hTBEC phenotype. Cyclophillin B and FK-associated binding protein, calcineurin A (alpha and beta), and NFATC3 and NFAT5 were detected in hTBEC cultures by RT-PCR. Acute and chronic cyclosporine treatment 1000 ng/mL significantly inhibited hTBEC proliferation, while tacrolimus did not (range of 10 ng/mL to 1000 ng/mL for acute treatment, 50 ng/mL for chronic treatment). Cyclosporin at 10 000 ng/mL significantly increased LDH release by well-differentiated hTBEC cultures (n = 6) and trended towards significance at 1000 ng/mL. IL1-beta stimulated IL-8 production was significantly increased in rapidly growing hTBEC cultures (n = 8) treated with cyclosporin (p = 0.049). Prolonged treatment of well-differentiated hTBECs at air-liquid-interface (ALI) with cyclosporin 1000 ng/mL significantly reduced intact multilayered mucociliary epithelium (p = 0.009). Inhibition of hTBEC growth, stimulation of IL-8 production and long-term effects on mucociliary phenotype and intact multi-layered epithelium suggest that cyclosporin may have a direct toxic effect on airway epithelium after transplantation.