干扰素-γ或IP-10对培养的HaCaT细胞增殖及炎症因子的影响

Effect of IFN-γ or IP-10 on Proliferation and Inflammation in Cultured HaCaT Cells

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。

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 cuhured keratinocytes. The immunoblotting of DNMT1 ,NF-kBp65,HDAC1 ,Cyclin D1, IL-6 or P16, specific MSP of P16 and immunostaining of NF-KBp65 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-KBp65, HDAC1 ,Cyclin D1 or IL-6 were up-regulated and the expression of PI6 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-KBp65 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-3, 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-IO/IFN-γ.