摘要
目的研究化合物1487B对肿瘤坏死因子-α(TNF-α)的作用,探讨其抗炎机制。方法利用荧光标记底物,检测在不同浓度化合物1487B作用下,THP-1细胞中肿瘤坏死因子-α转化酶(TACE)的活性;采用实时定量PCR法,分析肿瘤坏死因子-α的转录水平;利用酶联免疫吸附实验及免疫印迹,测定肿瘤坏死因子-α的分泌表达水平。结果在THP-1细胞中,1487B对胞内TACE酶活性有一定的抑制作用,实时定量PCR检测1487B可降低肿瘤坏死因子-α转录,且呈明显的剂量依赖性;免疫印迹及酶联免疫吸附实验检测终浓度为50、100、200μmol·L-1的1487B作用THP-1细胞后,肿瘤坏死因子-α前体的表达减少63%(P<0.01)、72%(P<0.01)、67%(P<0.01),肿瘤坏死因子-α的分泌水平降低24%(P<0.05)、31%(P<0.01)、70%(P<0.001)。结论1487B通过抑制THP-1细胞胞内TACE酶活性和肿瘤坏死因子-α转录,减少肿瘤坏死因子-α生成,本实验初步阐明了化合物1487B的抗炎机制,为其进一步应用奠定了基础。
OBJECTIVE To investigate the anti-inflammatory mechanism of compound 1487B by studying it with TNF-αexpression.METHODS After THP-1 treated with different concentrations of 1487B,the activity of TNF-αconverting enzyme(TACE)was measured with the special fluorescent peptide as substrates,the level of TNF-αmRNA was determined by Real-time qPCR,the secretion and expression levels of TNF-αwere detected through ELISA and Western blotting respectively.RESULTS The results showed that the activity of TACE and transcriptional level of TNF-αwere compromised by 1487 B addition with dose-dependent manner.The1487B(50,100,200μmol·L-1)can down-regulate 63%(P〈0.01),72%(P〈0.01),67%(P〈0.01)expression level of TNF-αprecursor and 24%(P〈0.05),31%(P〈0.01),70%(P〈0.001)secretion of TNF-αrespectively.CONCLUSION This study demonstrates that 1487B can block TNF-αproduction through both transcription and posttranscriptional mechanisms,illuminates its anti-inflammatory mechanisms and laid the foundation for compound to clinics.
作者
马明
张洋
郭连宏
白利平
姜蓉
李元
MA Ming;ZHANG Yang;GUO Lian-hong;BAI Li-ping;JIANG Rong;LI Yuan(Key Laboratory of Biotechnology of Anti-bitics,Ministry of Healsh,Institute of Medicinal Biotechnology,Chinese Academy of Medical Seiences&Peking Union Medical College,Beijing 100050,China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2015年第20期1806-1810,共5页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(31070086)
北京协和医学院协和青年科研基金资助项目(33320140093)
