麻黄碱对IL-17诱导的人永生化角质形成细胞HaCaT分泌CCL20的影响

摘要

目的 观察麻黄碱对IL-17诱导的人永生化角质形成细胞HaCaT分泌CCL20的影响.方法 将HaCaT细胞随机分为6组.A组加不含药物的培养液,B组加含20 ng/mL IL-17的培养液,C、D、E分别加含20 ng/mL IL-17联合800、1200、1600 μg/mL麻黄碱的培养液,F组加含20 ng/mL IL-17联合50 μg/mL MTX的培养液.培养36h后,采用ELISA法测定细胞上清液CCL20,采用RT-PCR法测定细胞中的CCL20 mRNA.结果 A、B、C、D、E、F组HaCaT细胞上清液CCL20水平分别为(97.72±1.86)、(159.43±7.09)、(147.05±2.25)、(138.60±8.04)、(126.66±7.04)、(114.39±3.90) pg/mL,HaCaT细胞CCL20 mRNA相对表达量分别为0.91±0.08、7.22±0.32、4.95±0.33、2.79±0.19、1.79±0.49、1.80±0.47;B、C、D、E、F组均高于A组(P均＜0.05),D、E、F组均低于B组(P均＜0.05),C、D组均低于F组(P均＜0.05).结论 麻黄碱能有效抑制IL-17诱导的HaCaT细胞分泌CCL20,为麻黄治疗银屑病提供一定的理论依据.%Objective To observe the effect of ephedrine on CCL20 secretion of IL-17-induced immortalized human keratinocytes (HaCaT).Methods HaCaT cells were randomly divided into 6 groups.Cells in the group A were only treated by DMEM.Cells in the group B were treated with 20 ng/mL IL-17 nutrient solution.Cells in the groups C,D and E were treated with 800,1200 and 1600 μg/mL ephedrine medium including 20 ng/mL IL-17.Cells in the group F were treated with 20 ng/mL IL-17 combined with 50 μg/mL MTX culture medium.After 36 h,ELISA was performed to determine the expression of CCL20 protein.RT-PCR was used to assess the expression of CCL20 mRNA.Results The expression levels of CCL20 protein in the cultural supernatant of HaCaT cells of groups A,B,C,D,E and F were (97.72 ±1.86),(159.43 ±7.09),(147.05 ±2.25),(138.60 ±8.04),(126.66 ±7.04),(114.39 ±3.90) pg/mL,respectively.The relative CCL20 mRNA expression in HaCaT cells of groups A,B,C,D,E,F were 0.91 ±0.08,7.22 ±0.32,4.95 ±0.33,2.79 ±0.19,1.79 ±0.49,1.80 ±0.47,respectively.Groups B,C,D,E and F were higher than group A (all P＜0.05);groups D,E,F were lower than group B (all P＜0.05);groups C and D were lower than group F (all P＜0.05).Conclusion Ephedrine can inhibit the CCL20 secretion of IL-17-induced HaCaT cells,and it provides a theoretical basis for the treatment of psoriasis by ephedrine or even diaphoresis.