蛋白激酶在大鼠过敏性炎症中白细胞产生中性粒细胞趋化因子过程中的可能作用。

Possible roles of protein kinases in neutrophil chemotactic factor production by leucocytes in allergic inflammation in rats.

作者信息

Tanabe J, Watanabe M, Kondoh S, Mue S, Ohuchi K

机构信息

Department of Pathophysiological Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Miyagi, Japan.

出版信息

Br J Pharmacol. 1994 Dec;113(4):1480-6. doi: 10.1111/j.1476-5381.1994.tb17163.x.

DOI:10.1111/j.1476-5381.1994.tb17163.x

PMID:7889305

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1510506/

Abstract

In an air pouch-type allergic inflammation model in rats, leucocytes that had infiltrated into the pouch fluid collected 4 h after the antigen challenge produced proteinaceous chemotactic factors for neutrophils when they were incubated in the medium. 2. To clarify the mechanism of activation of the infiltrated leucocytes in producing these factors, the effects of protein kinase inhibitors on neutrophil chemotactic factor production were examined. 3. When the infiltrated leucocytes were incubated for 4 h in medium containing the non-selective protein kinase inhibitor K-252a (1-100 ng ml-1, 2.14-214 nM), the tyrosine kinase inhibitor genistein (1-50 micrograms ml-1, 3.7-185 microM), and the more selective protein kinase C inhibitor H-7 (5-100 micrograms ml-1, 13.7-274 microM); neutrophil chemotactic activity in the conditioned medium was decreased in a concentration-dependent manner, but the adenosine 3':5'-cyclic monophosphate (cAMP)-dependent protein kinase inhibitor H-89 (1-1000 ng ml-1, 2.24-2240 nM) showed no effect. 4. Isoelectric focusing of the conditioned medium revealed that the leucocytes produced two neutrophil chemotactic factors, leucocyte-derived neutrophil chemotactic factor (LDNCF) 1 and LDNCF-2. Treatment of the leucocytes with K-252a, genistein, and H-7, but not H-89, inhibited production of both LDNCF-1 and LDNCF-2. 5. These results suggest that activation of tyrosine kinase and protein kinase C, but not cAMP-dependent protein kinase, is responsible for the production of LDNCF-1 and LDNCF-2. 6. The steroidal anti-inflammatory drug dexamethasone and the protein synthesis inhibitor cycloheximide inhibited neutrophil chemotactic factor production in a concentration-dependent manner. Time-course experiments showed that the inhibitory effect by dexamethasone was apparent even 30 min after the incubation.7. Mechanism for inhibiting the production of LDNCF-1 and LDNCF-2 by dexamethasone is also discussed.

摘要

在大鼠气囊型变应性炎症模型中，抗原攻击4小时后收集的气囊液中浸润的白细胞，在培养基中孵育时会产生针对中性粒细胞的蛋白质趋化因子。2. 为阐明浸润白细胞产生这些因子的激活机制，研究了蛋白激酶抑制剂对中性粒细胞趋化因子产生的影响。3. 当浸润的白细胞在含有非选择性蛋白激酶抑制剂K-252a（1 - 100 ng/ml，2.14 - 214 nM）、酪氨酸激酶抑制剂染料木黄酮（1 - 50 μg/ml，3.7 - 185 μM）和更具选择性的蛋白激酶C抑制剂H-7（5 - 100 μg/ml，13.7 - 274 μM）的培养基中孵育4小时时，条件培养基中的中性粒细胞趋化活性以浓度依赖的方式降低，但腺苷3':5'-环磷酸（cAMP）依赖性蛋白激酶抑制剂H-89（1 - 1000 ng/ml，2.24 - 2240 nM）没有效果。4. 条件培养基的等电聚焦显示白细胞产生了两种中性粒细胞趋化因子，白细胞衍生的中性粒细胞趋化因子（LDNCF）1和LDNCF-2。用K-252a、染料木黄酮和H-7处理白细胞，但不包括H-89，可抑制LDNCF-1和LDNCF-2的产生。5. 这些结果表明，酪氨酸激酶和蛋白激酶C的激活而非cAMP依赖性蛋白激酶，负责LDNCF-1和LDNCF-2的产生。6. 甾体抗炎药地塞米松和蛋白质合成抑制剂环己酰亚胺以浓度依赖的方式抑制中性粒细胞趋化因子的产生。时间进程实验表明，地塞米松的抑制作用在孵育后30分钟就很明显。7. 还讨论了地塞米松抑制LDNCF-1和LDNCF-2产生的机制。