【摘要】 目的探讨核因子-κB(NF-κB)反义寡核苷酸转染体外培养的HaCaT细胞反义株HaCaT后,对紫外线(UV)诱导的HaCaT细胞反义分泌炎症因子白介素6(IL-6)的抑制作用。方法蛋白质印迹方法测定不同剂量中波紫外线(UVB)辐射HaCaT细胞反义后NF-κBp65的变化;逆转录-聚合酶链反应测定NF-κBp65反义寡核苷酸转染后p65mRNA表达的变化;酶联免疫吸附测定法测定NF-κBp65反义寡核苷酸转染后紫外线辐射的HaCaT细胞反义分泌IL-6水平。结果10、20、30mJ/cm2UVB辐照HaCaT细胞反义后均可显著促进NF-κBp65表达(P<0.05),不同浓度的NF-κBp65反义寡核苷酸转染后对30mJ/cm2UVB诱导的p65mRNA表达和IL-6分泌均有显著抑制作用(P<0.05)。结论脂质体介导的NF-κBp65反义寡核苷酸转染HaCaT细胞反义,可以抑制UV辐射诱导的HaCaT细胞反义产生IL-6,表明UV诱导HaCaT细胞反义产生IL-6可能是通过UV激活NF-κBp65的表达产生的。更多还原

【Abstract】 Objective To explore the effects of antisense NF-κB p65 oligodeoxynucleotides on UV-induced IL-6 secretion of keratinocytes. Methods NF-κB p65 oligodeoxynucleotides were transfected to a keratinocyte cell line, HaCaT cells, mediated by lipofectamine. The NF-κB p65 protein in the cells was measured by Western blot assay, the mRNA level of NF-κB p65 was detected by RT-PCR, and UV-induced IL-6 level was determined by ELISA pre- and post-transfection and/or UVB irradiation. Results The NF-κB p65 protein expression was significantly increased in UVB (10, 20, 30 mJ/cm2 ) irradiation groups as compared with that of the control groups (P < 0.05). The p65 mRNA and UVB-induced IL-6 were inhibited after the keratinocytes were transfected with p65 antisense oligodeoxynucleotides at the concentrations of 2, 4 and 8 μg/mL (P < 0.05). Conclusions UV-induced IL-6 secretion of keratinocytes might be mediated by NF-κB, and could be inhibited by NF-κB p65 antisense oligodeoxynucleotides.更多还原