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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 investigate the effect of calcitonin gene related peptide (CGRP) on the secretion of chemokine CCL27 by HaCaT cells and to explore its mechanism. Methods CGRP was used to culture HaCaT cells in vitro. The mRNA expression of CCL27 was determined by RT-PCR. The chemokine CCL27 in supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The effect of CGRP on p38 mitogen-activated protein The role of kinase (p38 MAPK) phosphorylation. The p38 MAPK inhibitor SB203580 and the CGRP receptor antagonist CGRP8-37 were pretreated to observe the effect of CGRP on the secretion of the chemokine CCL27 in the supernatant. Results After CGRP incubation, the mRNA expression of CCL27, a chemokine, increased in HaCaT cells and increased in supernatant. CGRP induced phosphorylation of p38 MAPK, while SB203580 and CGRP8-37 inhibited this effect and partially inhibited the secretion and mRNA expression of chemokine CCL27 in CGRP-induced supernatant. Conclusion CGRP can induce chemotaxis of CCL27 in HaCaT cells, and its mechanism of action depends partly on the phosphorylation of p38 MAPK.