摘要
目的:观察肿瘤坏死因子-α(TNF-α)对瘢痕疙瘩成纤维细胞(KFB)以及正常皮肤成纤维细胞(NSF)核因子-kappa B(NF-κB)的影响,以探讨瘢痕疙瘩的发生机制。方法:原代培养成纤维细胞;免疫荧光技术观察NF-κB p65和IκB-α在静息状态和TNF-α刺激后的成纤维细胞中分布;应用TransAMTM NF-κB p65 kit试剂盒检测NF-κB p65 DNA结合活性;应用Western blotting检测IκB-α蛋白水平。结果:TNF-α刺激后,NF-κB p65从细胞浆转移至细胞核;NF-κB p65 DNA结合活性水平在刺激后1h达到高峰,4h接近正常;细胞浆IκB-α蛋白水平在刺激后15min降至最低值,4h基本接近正常;KFB较NSF对TNF-α的刺激更为敏感。结论:KFB较NSF对TNF-α活化NF-κB更为敏感,可能是瘢痕疙瘩形成的潜在发病机制。
AIM： To investigate NF - κB p65 activation and IκB - α expression in keloid fibroblasts （KFB） and normal skin fibroblasts （NSF） stimulated with TNF - α and to explore the underlying molecular pathogenesis of keloid formation. METHODS ： Primary KFB was cultured. The location of NF - κB p65 and IκB - α in KFB and NSF at quiescent condition and the nuclear translocation of NF - κB p65 after TNF - α stimulation were observed by immunofluorescence technique, NF - κB p65 DNA binding activity was detected with TransAM^TM NF - κB p65 kit. The IKB - α protein level was determined by means of Western blotting technique. RESULTS ： After stimulated with TNF - α, NF - κB p65 translocated into the nucleus. NF - κB p65 DNA binding activity increased to its maximum at 1 h and was dropped to normal at 4 h. TNF - α induced most degradation of IκB - α at 15 min and became detectable in cytoplasm after 4 h. KFB showed more sensitive ability to TNF - α stimulation than NSF. CONCLUSION ： NF - κB may play a role in keloid pathogenesis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第1期165-168,共4页
Chinese Journal of Pathophysiology
基金
山东省自然科学基金资助项目(No.Q2006C09)
