CAJ | 학술논문

目的：探讨红霉素对香烟烟雾刺激的人巨噬细胞组蛋白去乙酰化酶3（HDAC3）表达的影响。方法：体外培养人类单核细胞系U937细胞，用佛波脂（PMA）将其诱导分化为人巨噬细胞。按传统方法制备好香烟烟雾提取物（CSE）用于实验。将传代后的细胞分组：对照组、CSE组、红霉素＋CSE组、HDAC特异性抑制剂曲古霉素（ TSA）组。采用比色法检测细胞HDAC总活性；Western blot检测HDAC3和NF-κB蛋白表达；通过酶联免疫吸附实验（ ELISA ）检测细胞上清液中TNF-α的浓度。结果：1％CSE刺激人巨噬细胞引起细胞HDAC总活性减弱和HDAC3蛋白表达下降，诱导转录因子NF-κB活性增高，释放TNF-α；红霉素（1μg/ml）预孵育24 h可以增强CSE抑制的巨噬细胞HDAC3蛋白表达，下调NF-κB蛋白表达，抑制TNF-α的合成和释放。结论：红霉素通过上调HDAC3蛋白表达水平而抑制香烟烟雾诱导增强的转录因子NF-кB活性，进而影响炎症介质TNF-α的合成调控。
목적：탐토홍매소대향연연무자격적인거서세포조단백거을선화매3（HDAC3）표체적영향。방법：체외배양인류단핵세포계U937세포，용불파지（PMA）장기유도분화위인거서세포。안전통방법제비호향연연무제취물（CSE）용우실험。장전대후적세포분조：대조조、CSE조、홍매소＋CSE조、HDAC특이성억제제곡고매소（ TSA）조。채용비색법검측세포HDAC총활성；Western blot검측HDAC3화NF-κB단백표체；통과매련면역흡부실험（ ELISA ）검측세포상청액중TNF-α적농도。결과：1％CSE자격인거서세포인기세포HDAC총활성감약화HDAC3단백표체하강，유도전록인자NF-κB활성증고，석방TNF-α；홍매소（1μg/ml）예부육24 h가이증강CSE억제적거서세포HDAC3단백표체，하조NF-κB단백표체，억제TNF-α적합성화석방。결론：홍매소통과상조HDAC3단백표체수평이억제향연연무유도증강적전록인자NF-кB활성，진이영향염증개질TNF-α적합성조공。
Objective:To study the effect of erythromycin(EM) on cigarette smoke-induced histone deacetylase-3(HDAC3) protein expression in human macrophages in vitro .Methods:The Aqueous cigarette smoke extract ( CSE) was always prepared fresh on the day of the experiment .The U937 monocytic cells were differentiated into macrophages by using phorbol 12-myristate 13-acetate (PMA) according to standard procedures .The U937 differentiated cells were treated with either CSE (1%) or EM (1 μg/ml) pre-treatment, and HDAC inhibitor trichostatin A (TSA;100 ng/ml) for 24 h.HDAC activity was measured with a colorimetric assay kit and Western blot was used for HDAC3 and factor nuclear-kappaB (NF-κB) protein assays.The levels of tumor necrosis factor-α(TNF-α) release in the supernatant were determined by enzyme linked immunosorbent assay (ELISA).Results:CSE(1%) significantly de-creased HDAC activity and HDAC 3 protein levels at 24 h.Preincubation with EM (1μg/ml ) for 24 h significantly inhibit CSE (1%) induced decrease of HDAC3 protein expression.Furthermore, Preincubation with EM(1 μg/ml) for 24 h significantly inhibit NF-κB activity and TNF-αrelease in human macrophages .Conclusion:EM is able to restore HDAC3 levels decreased by cigarette smoke and inhibit NF-κB activity resulting in decreasing CSE-mediated TNF-αrelease, which has shown an important explanation that EM possess the anti-inflammatory effect induced by cigarette smoke .