咪喹莫特对朗格汉斯细胞及角质形成细胞细胞因子mRNA表达及分泌水平的影响

Effects of Imiquimod on the mRNA expression and secretion of cytokines by human epidermal Langerhans cells and keratinocytes

目的观察咪喹莫特(Imiquimod)溶液对人表皮朗格汉斯细胞(LC)及永生化角质形成细胞(KC)株-HaCaT细胞细胞因子mRNA及分泌水平的影响,探讨其免疫调节机制。方法联用密度梯度离心及免疫磁珠法分离纯化人表皮LC,将LC及HaCaT细胞均各分为实验组和对照组,实验组加5μgml咪喹莫特,对照组不加药,处理4h后,提取总RNA,采用RTPCR法分别检测各组细胞中细胞因子TNFα、IL1β及IL6mRNA的表达量,取培养上清,用ELISA方法检测3种细胞因子的含量。结果LC实验组TNFα、IL1β及IL63种细胞因子mRNA表达量及分泌量均较对照组显著增高(P<0.05),而HaCaT细胞组加药后3种细胞因子mRNA表达量及分泌量较对照组未见明显增加(P>0.05)。结论5μgml咪喹莫特溶液能增加LC细胞因子TNFα、IL1β及IL6的mRNA表达量,但其对HaCaT细胞上述3种细胞因子mRNA表达量均未见有明显上调作用。

Objective To observe the effects of the Imiquimod solution on the mRNA expressions of cytokines（TNF-α, IL-1β and IL-6） and cytokine secretion from human epidermal Langerhans cells and human eternal cell lines--HaCaT kerafinocyte to illuminate the immunoregulation mechanisms of Imiquimod. Methods Langerhans cells were sorted by density gradient centrifugation and magnetic technique. Langerhans cells and HaCaT cells were divided into experimental group and control group. The solution of 5μg/ml Imiquimod was added to the experimental group. The total BNA and cell-free culture supematants were harvested 4 hours after application. BTPCR was conducted to detect the mRNA expressions of TNF-α, IL-1β and IL-6 in each group, cytokines secretion was determined by ELISA kits. Results Four Four after 5μg/ml Imiquimod application, expression levels of TNF-α, IL-1β and 1I.-6 mRNA and cytokines secretion from Langerhans cells were statistically higher than those of controls（ P 〈 0.05）. On the contrary, there was no statistically difference of the mRNA expression levels and cytokine secretion of TNF-α, IL-1β and IL-6 of HaCaT cells between the experimental group and the control group（ P 〉0.05）. Conclusion The solution of 5μg/ml Imiquimod can increase the mRNA expression and secretion of TNF-α, ID-1β and IL-6 in sorted Langerhans cells. But in the HaCaT cells, Imiquimod fags to increase the mRNA expression and secretion of the above cytokines.