【摘要】 目的:通过研究干扰素(IFN-α)对Jurkat E6-1细胞内CCR5 mRNA表达及基因表达调控的影响,探讨采用干扰素治疗相关疾病的方法。方法:用不同浓度的IFN-α刺激处于对数生长期的CD4+T淋巴细胞系Jurkat E6-1细胞。培养48小时后,提取细胞的总RNA,逆转录成cDNA,进行RT-PCR和Real time-PCR扩增目的基因CCR5;利用脂质体转染荧光素酶报告系统检测Jurkat E6-1细胞内CCR5活性变化情况。结果:(1)IFN-α在浓度为100 U/ml时对CCR5 mRNA的表达有明显抑制作用;在浓度为1 000 U/ml时对CCR5 mRNA的表达表现为增强作用;在浓度为10 000 U/ml时对CCR5 mRNA的表达增强作用有所减弱。(2)当IFN-α的浓度为100 U/ml时荧光素酶的活性最低;当IFN-α的浓度为1 000 U/ml时荧光素酶的活性最高;当IFN-α的浓度为10 000 U/ml时荧光素酶的活性又有所减低。结论:IFN-α作为一类很好的免疫调节、抗病毒生物制剂,不同剂量在一定范围内会对细胞CCR5的表达有显著影响。更多还原

【Abstract】 Objective:To discuss the treatment methods of related diseases with IFN-α by the study of effects of IFN-α on the expression and regulation of CCR5 in Jurkat E6-1 cells.Methods:The Jurkat E6-1 cells in logarithmic growth phase were stimulated by IFN-α with different concentrations.After incubation for 48 h,total cellular RNA was isolated and reverse-transcribed into cDNA.Then PCR analysis were performed to detect CCR5 and β-actin(as an internal standard) gene expression.Then luciferase enzymatic activity detection were performed by a CCR5 luciferase reporter plasmid using the TfxTM-50 Reagent(Promega).Results:(1) 100 U/ml IFN-α could inhibited CCR5 expression.But,1 000 U/ml IFN-α could promoted CCR5 expression and the promotion effect of 10 000 U/ml IFN-α was lower than 1 000 U/ml IFN-α.(2) The activity of luciferase was the lowest at 100 U/ml IFN-α,but the activity of luciferase was the highest at 1 000 U/ml IFN-α.However,the activity of luciferase reduce at 10 000 U/ml IFN-α.Conclusion:IFN-α,conducted as immunoregulation and antivirus biological agent,could effect CCR5 expression significantly with different concentration.更多还原