Modulation of endothelin-1 production by a pulmonary epithelial cell line. I. Regulation by glucocorticoids.
Modulation of endothelin-1 production by a pulmonary epithelial cell line. I. Regulation by glucocorticoids.
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肺上皮细胞系对内皮素-1 产生的调节。
DOI：
10.1016/0006-2952(94)90506-1
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发表时间：
1994
影响因子：
5.8
通讯作者：
Szentivanyi,A
中科院分区：
文献类型：
--
作者：
Calderón,E;Gómez-Sánchez,CE;Cozza,EN;Zhou,M;Coffey,RG;Lockey,RF;Prockop,LD;Szentivanyi,A
内皮素-1（ET-1）是迄今为止所描述的最有效的支气管收缩剂之一。哮喘患者的支气管上皮细胞在体内表达ET-1前体，在体外释放大量的ET-1。健康和慢性支气管炎对照不表达preproET-1或释放ET-1。白细胞介素-2（IL-2）等细胞因子可上调豚鼠气道上皮细胞ET-1的释放。由于ET-1可能在哮喘的发病机制中起基础作用，我们探讨了两种糖皮质激素地塞米松（Dex）和曲安奈德（TA）是否抑制了转化的人肺上皮细胞系A549细胞合成和释放ET-1。细胞在RPMI 1640加10%胎牛血清（FBS）中生长至汇合。然后将细胞在无血清条件下培养3天以获得ET-1基础水平。用放射免疫法（RIA）测定10%FBS、IL-2（10 U/mL）、地塞米松（Dex）、TA和米非司酮（1、10或100 nM）对ET-1的影响。在对照培养物中，ET-1的产生从6小时的57.6 ± 5 pg/mg细胞蛋白增加到72小时的170 ± 9 pg/mg细胞蛋白。10%FBS使ET-1的产生从58.7 ± 9.6增加到399 ± 14.5pg/mg细胞蛋白。IL-2显著增加ET-1，24小时从100.7 ± 6.1增加到144 ± 6.7，72小时从170 ± 9增加到207.7 ± 24。这两种药物（仅在100 nM）减少ET-1的生产在10%的FBS和IL-2刺激的细胞。米非司酮（10和100 nM）可逆转地塞米松（100 nM）在24-72 h诱导的ET-1产生的减少，北方印迹分析显示地塞米松（100 nM）在6和24 h降低ET-1 mRNA的表达，而米非司酮（100 nM）可逆转地塞米松诱导的细胞ET-1产生的减少。总之，地塞米松和TA下调合成和生产的ET-1的人肺上皮细胞系在基础或刺激条件下，这些影响被逆转米非司酮。这些发现提示糖皮质激素在哮喘治疗中的作用是一种新的机制。
Endothelin-1 (ET-1) is one of the most potent bronchoconstrictor agents yet described. Bronchial epithelial cells of asthmatic patientsin vivoexpress preproET-1 andin vitrorelease high amounts of ET-1. Healthy and chronic bronchitic controls do not express preproET-1 or release ET-1. Interleukin-2 (IL-2) and other cytokines up-regulate thein vitroET-1 release in guinea pig airway epithelial cells. We explored whether two glucocorticoids, dexamethasone (Dex) and triamcinolone acetonide (TA), inhibit the synthesis and release of ET-1 by A549 cells, a transformed human pulmonary epithelial cell line, since ET-1 may have a basic role in the pathogenesis of asthma. Cells were grown to confluence in RPMI 1640 plus 10% fetal bovine serum (FBS). Cells were then cultured for 3 days without serum to obtain ET-1 basal levels. The effects of 10% FBS, IL-2 (10 U/mL), Dex, TA or mifepristone, a steroid antagonist (1, 10 or 100 nM), were evaluated on ET-1 as measured by radioimmunoassay (RIA). ET-1 production increased from 57.6 ± 5 pg/mg cell protein at 6 hr to 170 ± 9 pg/mg cell protein at 72 hr in control cultures. Ten percent FBS increased ET-1 production from 58.7 ± 9.6 to 399 ± 14.5 pg/mg cell protein. IL-2 significantly increased ET-1 from 100.7 ± 6.1 to 144 ± 6.7 at 24 hr and from 170 ± 9 to 207.7 ± 24 at 72 hr. Dex and TA (10 and 100 nM) at 24–72 hr decreased ET-1 under basal conditions. Both drugs (only at 100 nM) decreased ET-1 production in 10% FBS- and IL-2-stimulated cells. Mifepristone (10 and 100 nM) reversed the decreased production of ET-1 induced by Dex (100 nM) at 24–72 hr. Northern blot analysis showed that Dex (100 nM) decreased the expression of ET-1 mRNA at 6 and 24 hr, but that mifepristone (100 nM) reversed this effect in cells cultured with Dex. In conclusion, Dex and TA down-regulated the synthesis and production of ET-1 by this human pulmonary epithelial cell line under basal or stimulated conditions, and these effects are reversed by mifepristone. These findings suggest a novel mechanism of glucocorticoid effect during the treatment of asthma.