摘要
目的了解人肥大细胞系(humanmastcellline,HMC1)在佛波醇酯(phorbol12myristate13acetate,PMA)+钙离子通道(A23187)激发下能否产生白介素(Interleukin,IL)4,并且观察布地奈得、地氯雷他定和地塞米松等药物对IL4的产生和表达的影响。方法HMC1在10-6~10-10mol/L浓度的药物中孵育1h后,以25ng/mlPMA+25×10-7mol/LA23187激发细胞因子生成,再分别孵育6、12或24h,提取上清液,采用酶联免疫吸附法(enzymelinkedimmunosorbentassays,ELISA)测定IL4水平,并采用半定量逆转录聚合酶链反应(reversetranscriptionpolymerasechainreaction,RTPCR)观察药物对IL4mRNA表达水平的影响。结果HMC1在激发后可产生IL4,IL4mRNA的表达也很明显。布地奈得、地氯雷他定和地塞米松在各时间点和浓度下,均对HMC1IL4的产生有明显的抑制作用(同未加药物的对照组比较,P值均<005),抑制率在(361±40)%到(679±38)%之间。布地奈得和地塞米松对IL4mRNA的表达无抑制作用,而地氯雷他定则能明显抑制IL4mRNA的表达水平。结论HMC1经激发可表达和产生IL4,布地奈得、地氯雷他定和地塞米松能显著抑制IL4的产生,地氯雷他定还能抑制IL4mRNA的表达。
Objective Since human mast cell is an important source of cytokines, it is of importance to understand the effects of anti-allergic drugs on cytokines modulation in mast cells. In the present study, we aimed at observing whether IL-4 could be released from human mast cell line (HMC-1) after the stimulation of PMA+A23187, and the effects of systemic glucocorticosteroid, dexamethasone, topical glucocorticosteroid, budisonide and H1 antagonist, desloratadine on IL-4 release and mRNA expression. Methods HMC-1 was stimulated with 25 ng/ml phorbol 12-myristate 13-acetate (PMA) and 2.5× 10^-7 mol/L ionomycin (A23187) and cultured for 6 hours, 12 hours and 24 hours respectively in the presence or absence of 10^-6 -10^-10 mol/L concentrations of test drugs. Culture supernatants were collected and the levels of IL-4 were assayed by enzyme-linked immunosorbent assays (ELISA). The mRNA expression of IL-4 was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Results HMC-1 expressed IL-4 mRNA and the resulting protein production of IL-4 released after being stimulated with PMA plus A23187. Dexamethasone, budesonide and desloratadine had potent inhibitory effect on IL-4 release at any concentrations and time points, with significant deference (P<0.05) compared to the control cells. The inhibitory effect did not show time-dependent and concentration-dependent manner. Desloratadine and budesonide showed neither up-regulatory nor down-regulatory effects on IL-4 mRNA expression at the test concentrations, however, desloratadine could down-regulate IL-4 mRNA expression. Conclusions HMC-1 could express and produce IL-4 after stimulation. Dexamethasone, budesonide and desloratadine all had inhibitory effects on IL-4 release from HMC-1.In addition, desloratadine could also inhibit the IL-4 mRNA expression.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2005年第2期124-127,共4页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
