p38 MAPK介导降钙素基因相关肽刺激HaCaT细胞分泌CCL27的研究

p38 MAPK mediates production of CCL27 by calcitonin gene-related peptide-stimulated HaCaT cells

目的观察降钙素基因相关肽(CGRP)对HaCaT细胞珠分泌趋化因子CCL27的影响,并探讨其作用机制。方法以CGRP孵育体外培养的HaCaT细胞,应用RT-PCR方法测定CCL27的mRNA表达,酶联免疫吸附试验(ELISA)测定上清液中趋化因子CCL27,Western blot测定CGRP对p38丝裂原活化蛋白激酶(p38 MAPK)磷酸化的作用。应用p38 MAPK抑制剂SB203580及CGRP受体拮抗剂CGRP8-37预处理,观察CGRP对上清液中趋化因子CCL27分泌的影响。结果应用CGRP孵育后,HaCaT细胞的趋化因子CCL27的mRNA表达上升,上清液中分泌量增加。CGRP诱导p38 MAPK磷酸化,而SB203580及CGRP8-37可抑制这一作用,并部分抑制CGRP诱导的上清液中趋化因子CCL27的分泌及其mRNA表达。结论 CGRP可引起HaCaT细胞分泌趋化因子CCL27,其作用机制部分依赖于p38 MAPK的磷酸化。

Objective To examine whether calcitonin gene-related peptide（CGRP） can induce the production of chemotactic cytokines ligand 27（CCL27） by the human keratinocyte cell line HaCaT,and to identify the mechanism of the signal pathway involved in the effect of CGRP.Methods Expression of CCL27 mRNA in CGRP-stimulated cultural HaCaT cells was analyzed by reverse transcription PCR（RT-PCR）.The concentration of CCL27 in the supernatants was measured by the ELISA method.Protein expression levels of phosphorylated p38 MAPK（p-p38 MAPK） were detected by Western blotting.Results Incubation with CGRP increased not only mRNA expressions and protein levels of CCL27 in supernatants,but also expression of p-p38 MAPK.Pretreatment with SB203580 and CGRP8-37 decreased CGRP-induced expression of p-p38 MAPK and the CCL27 mRNA level.SB203580 and CGRP8-37 decreased the concentration of CCL27 in supernatants.Conclusion CGRP can induce CCL27 secretion of HaCaT cells through p38 MAPK phosphorylation.