去甲肾上腺素抑制小鼠真皮微血管内皮细胞CXCL10和CCL17表达

Norepinephrine Inhibits CXCL10 and CCL17 Expressions in Mouse Dermal Microvascular Endothelial Cells

目的通过研究去甲肾上腺素(NE)对小鼠真皮微血管内皮细胞(pDMECs)趋化因子CXCL10和CCL17表达的影响,探讨NE对炎症性皮肤病的调控机制。方法采用不同浓度NE对脂多糖(LPS)诱导的小鼠pDMECs进行干预,ELISA法检测CXCL10和CCL17的表达;分别用α肾上腺素受体拮抗剂酚妥拉明(Phent)或β肾上腺素受体拮抗剂普萘洛尔(Prop)封闭受体,再予NE进行干预,ELISA法检测小鼠pDMECs CXCL10和CCL17的表达,明确NE作用的受体类型。BALB/c小鼠背部皮内注射NE,LPS诱导皮肤炎症,24 h后取小鼠背部真皮组织,实时荧光定量聚合酶链反应(qRT-PCR)检测CXCL10和CCL17 mRNA的表达水平,体内实验进一步验证NE参与皮肤炎症调控。结果LPS诱导的小鼠pDMECs CXCL10和CCL17表达较对照组明显增加,不同浓度NE抑制LPS诱导的小鼠pDMECs CXCL10和CCL17表达,呈剂量依赖性。小鼠pDMECs预先给予Prop处理后,削弱了NE对LPS诱导CXCL10和CCL17表达的抑制效应;预先给予Phent处理后,NE对LPS诱导CXCL10和CCL17表达的抑制作用仍存在。经LPS诱导的BALB/c小鼠,NE皮内注射组真皮CXCL10和CCL17 mRNA的表达水平较PBS注射组明显降低。结论NE可通过皮肤血管内皮细胞上的β肾上腺素受体,抑制趋化因子CXCL10和CCL17的表达,参与皮肤炎症调控。

Objective To investigate the regulatory mechanism of norepinephrine(NE)on inflammatory skin diseases,we study the effect of NE on the expressions of chemokines CXCL10 and CCL17 in primary dermal microvascular endothelial cells(pDMECs)derived from BALB/c mice.Methods Different concentrations of NE on LPS-induced chemokines CXCL10 and CCL17 expressions by pDMECs were investigated by sandwich ELISA.To determine whether the effects of NE were mediated byαorβadrenergic receptors,pDMECs were treated with phentolamine(Phent)or propranolol(Prop)followed by the addition of NE and LPS.The concentrations of CXCL10 and CCL17 in supernatants were quantified by ELISA.To verify the regulation effects of NE on skin inflammation in vivo,BALB/c mice were injected intradermally on the dorsum with NE prior to administration of LPS,and the expression levels of CXCL10 and CCL17 mRNA on dermis were detected by real time fluorescent quantitative polymerase chain reaction(qRT-PCR)after 24 h.Results The expressions of CXCL10 and CCL17 in LPS-induced mice pDMECs were significantly increased compared with the control group,however,different concentrations of NE inhibited the expressions of CXCL10 and CCL17 in LPS-induced mice pDMECs in a dose-dependent manner.The inhibitory effect of NE on LPS-induced CXCL10 and CCL17 expressions were attenuated by pre-treatment of pDMECs withβ-adrenoceptor antagonist Prop,while remained after pre-treatment withα-adrenoceptor antagonist Phent.In vivo,levels of CXCL10 and CCL17 mRNA in the dermis of LPS-induced BALB/c mice were significantly decreased in NE injection group compared with PBS injection group.Conclusion NE exhibits significant regulatory effects on skin inflammation through CXCL10 and CCL17 production,which may mediate primarily viaβadrenergic receptor on dermal microvascular endothelial cells.