【摘要】 目的探讨以干扰素-γ(IFN-γ)或IFN-γ诱导蛋白10(IP-10)培养的HaCaT细胞的特点并与银屑病皮损作比较。方法用IP-10(20ng/mL),IFN-γ(50U/L)处理HaCaT细胞进行48h培养;收集寻常性银屑病患者皮损及正常包皮作为对照;提取各细胞和组织的蛋白质及基因组DNA;免疫印迹检测DNMT1,NF-κBp65,HDAC1,CyclinD1,IL-6及P16的表达,基因组DNA进行P16抑癌基因的甲基化特异PCR(MSP)并对各组标本进行NF-κBp65及IL-6的免疫染色。结果免疫印迹显示IP-10组、INF-γ组与病变组织及各相应对照组相比结果相似,DNMT1,NF-κBp65,HDAC1,CyclinD1及IL-6表达均上调,P16表达皆下调,差异有统计学意义(P<0.05)。MSP结果显示,IP-10组、IFN-γ组与皮损组织的甲基化P16皆高于各对照组,差异有统计学意义(P<0.05)。免疫染色结果显示,NF-κBp65及IL-6的表达均强于各对照组,差异有统计学意义(P<0.05)。结论 IP-10与IFN-γ可显著提高HaCaT细胞的增殖及炎症因子的表达,并引起相关表现遗传学的改变,提示IP-10与IFN-γ可能参与银屑病的发生机制与发展,以IP-10/IFN-γ培养的HaCaT细胞模拟了部分银屑病的特点。更多还原

【Abstract】 Objective To compare the cultured karatinocytes induced by IP-10/IFN-γ and characteristics of psoriatic lesions . Methods The HaCaT cells were cultured and treated with IP-10(20ng/mL) ,IFN-γ(50U/L) for 48h. The total protein and genome DNA were extracted from lesion of psoriasis vulgaris,healthy people forskin and each group of cultured keratinocytes. The immunoblotting of DNMT1,NF-κBp65,HDAC1,Cyclin D1,IL-6 or P16,specific MSP of P16 and immunostaining of NF-κBp65 and IL-6 in each group samples were performed. Results The immunoblotting showed that when compared with respective corresponding controls,IP-10/INF-γ groups and lesion groups were similar. The expression of DNMT1,NF-κBp65, HDAC1,Cyclin D1 or IL-6 were up-regulated and the expression of P16 was down-regulated,P < 0. 05. The MSP of P16 showed that the methylated p16 was higher than that in the control,P < 0. 05. The immunostaining of NF-κBp65 and IL-6 was intenser in IP-10/IFN-γ and lesion groups,when compared with corresponding control groups,P < 0. 05. Conclusion IP-10 or IFN-γ can promote proliferation and inflammatory cytokine in HaCaT cells. IP-10/IFN-γ might be correlated to the occurrence and develop of psoriasis. It is suggested that the part of psoriatic keratinocytes model may be established through co-cultured keratinocytes with IP-10/IFN-γ.更多还原