Establishment of an in vitro test system to evaluate the down-regulatory activities of natural products on IL-4
Establishment of an in vitro test system to evaluate the down-regulatory activities of natural products on IL-4
复制标题
DOI：
10.1007/bf02980244
复制
发表时间：
2007-09-01
影响因子：
6.7
通讯作者：
Jin, Mirim
中科院分区：
文献类型：
--
作者：
Choi, Jeong June;Park, Bo-Kyung;Jin, Mirim
白细胞介素-4 (IL-4) 是一种代表性的 T(H)2 细胞因子，在特应性皮炎、特应性鼻炎和哮喘等多种过敏性疾病的发病中发挥病理作用。已经研究了几种下调 IL-4 表达的候选药物在临床环境中作为抗过敏剂的可能性。因此，评估 IL-4 启动子活性的体外测试可能有助于选择新型天然疗法的候选者。以潮霉素抗性基因作为选择标记，将IL-4的启动子区(741至+56)克隆到质粒pGL4.14中的荧光素酶基因上游，产生pGL4.14-IL-4。在瞬时转染 pGL4.14-IL-4 的 EL-4 胸腺瘤和 RBL-2H3 细胞以及持续表达 pGL4.14-IL-4 的稳定细胞系中，用 PMA 和 A23187 处理后，荧光素酶活性比对照显着增加约 10 倍。众所周知的抗过敏药物环孢素 A 和地塞米松以剂量依赖性方式显着下调活性。通过测量 TBRC 植物文库的各种提取物对 PMA 和 A23187 诱导的 IL-4 启动子荧光素酶活性的下调活性，并使用 ELISA 测定测量培养细胞中 IL-4 的产生，评估了该系统的可行性。本研究的结果表明，该初步筛选系统简单且节省时间，并且可能适合通过测量天然产物对 IL-4 启动子的下调作用来选择过敏性疾病的天然治疗候选物。
Interleukin-4 (IL-4), a representative T(H)2 cytokine, plays a pathologic role in the onset of various allergic diseases including atopic dermatitis, atopic rhinitis, and asthma. Several drug candidates that down-regulate IL-4 expression have been studied for their possible use as antiallergic agents in clinical settings. Therefore, an in vitro test to evaluate IL-4 promoter activities might be useful for selecting candidates of novel natural therapeutics. The promoter region (741 to +56) of IL-4 was cloned upstream of a luciferase gene in the plasmid pGL4.14 with a hygromycin resistance gene as a selection marker to generate pGL4.14-IL-4. Treatment with PMA and A23187 highly increased luciferase activity by approximately 10-fold compared with the control in both EL-4 thymoma and RBL-2H3 cells transiently transfected with pGL4.14-IL-4, as well as in stable cell lines constantly expressing pGL4.14-IL-4. Cyclosporin A and dexamethasone, well-known anti-allergic agents, significantly down-regulated the activity in a dosedependent manner. The feasibility of this system was evaluated by measuring the down-regulatory activities of various extracts from the TBRC plant library on PMA- and A23187-induced luciferase activities of IL-4 promoter, and by measuring IL-4 production in cultured cells using ELISA assays. The results of this study suggest that this primary screening system is simple and time-saving, and might be suitable for the selection of natural therapeutic candidates for allergic disease by measuring the down-regulatory effects of natural products on the IL-4 promoter.