卡泊三醇对TNF-α诱导的人角质形成细胞中CCL20表达的影响

Effect of calcipotriol on CCL20 expression induced by TNF-αin human keratinocytes

目的:明确卡泊三醇体外对TNF-α诱导的人角质形成细胞CCL20表达的影响。方法:体外培养人角质形成细胞,分为TNF-α组,卡泊三醇组,TNF-α+卡泊三醇组,TNF-α+卡泊三醇+SB202190(p38 MAPK抑制剂)组,TNF-α+卡泊三醇+SP600125(JNK抑制剂)组,TNF-α+卡泊三醇+U0126(ERK抑制剂)组,TNF-α+卡泊三醇+CAPE(NF-κB抑制剂)组。实时荧光定量PCR法和酶联免疫吸附试验检测CCL20 mRNA及蛋白表达水平。Western blot法检测p38 MAPK,ERK和NF-κBp65磷酸化情况。结果:TNF-α+卡泊三醇组CCL20的表达水平及ERK,p38 MAPK,NF-κBp65磷酸化水平低于TNF-α组,差异均有统计学意义(均P<0.05)。TNF-α+卡泊三醇组CCL20的表达水平低于TNF-α+卡泊三醇+U0126(ERK抑制剂)组,高于TNF-α+卡泊三醇+SB202190(p38 MAPK抑制剂)组和TNF-α+卡泊三醇+CAPE(NF-κB抑制剂)组,差异均有统计学意义(均P<0.05)。CCL20的表达水平在TNF-α+卡泊三醇组与TNF-α+卡泊三醇+SP600125(JNK抑制剂)组间比较差异无统计学意义(P>0.05)。结论:卡泊三醇可能通过调节TNF-α诱导的ERK,p38 MAPK,NF-κBp65磷酸化,从而参与调节人角质形成细胞中CCL20表达。

Objective:To deremine the effect of calcipotriol on the expression of CCL20 induced by TNF-αin human keratinocytes.Methods:The keratinocytes were cultured in vitro,and then,were divided into TNF-αgroup,calcipotriol group,TNF-α+calcipotriol group,TNF-α+calcipotriol+SB202190(p38 MAPK inhibitor)group,TNF-α+calcipotriol+SP600125(JNK inhibitor)group,TNF-α+calcipotriol+U0126(ERK inhibitor)group and TNF-α+calcipotriol+CAPE(NF-κB inhibitor)group.The expression level of CCL20 mRNA and protein was detected by real time-QPCR and ELISA.The phosphorylation status of p38 MAPK and ERK and NF-κBp65 was detected by Western blot.Results:The level of CCL20 and phosphorylation levels of p38 MAPK,ERK and NF-κBp65 in TNF-α+calcipotriol group were lower than in TNF-αgroup,with significant differences(P s<0.05).The level of CCL20 in TNF-α+calcipotriol group was lower than in TNF-α+calcipotriol+U0126(ERK inhibitor)group,higher than those in TNF-α+calcipotriol+SB202190(p38 MAPK inhibitor)group and CAPE(NF-κB inhibitor)group,with significant differences(P s<0.05).There was no significant difference in CCL20 level between TNF-α+calcipotriol group and TNF-α+calcipotriol+SP600125(JNK inhibitor)group.Conclusion:Calcipotriol can inhibit the CCL20 expression induced by TNF-αin keratinocytes,which may be through regulating ERK,p38 MAPK and NF-κB p65 phosphorylation.