IL-31和他克莫司对Hacat细胞NT4表达的影响

Effects of IL-31 or tacrolimus on expression of NT4 in human Hacat cells

摘要:

目的：探讨白介素-31（interleukin-31，IL-31）、他克莫司对角质形成细胞神经营养素-4（neurotrophin-4，NT4）表达的影响。方法：培养人角质形成细胞株（Hacat细胞），采用不同浓度他克莫司作用24 h，噻唑蓝比色法（MTT法）检测细胞增殖活性。IL-31、他克莫司单独或共同作用于Hacat细胞24 h后，采用real-time PCR法检测NT4 mRNA表达，酶联免疫吸附法（ELISA）分别检测12、24 h各组细胞上清液中NT4含量。结果：低浓度他克莫司（10、100）ng/ml对细胞活性无明显影响（P >0.05），高浓度（1 000 ng/ml）时细胞活性受到抑制（P=0.002）。IL-31作用Hacat细胞后，其NT4mRNA表达和上清中NT4含量均增加（P<0.05）；100 ng/ml他克莫司作用24 h后他克莫司组和联合作用组细胞NT4 mRNA表达量和细胞上清中NT4含量均分别低于对照组和IL-31组（P<0.05）。结论：IL-31可促进角质形成细胞NT4表达，他克莫司可能通过降低角质形成细胞NT4表达水平而发挥其抗瘙痒作用。

Abstract:

Objective：To investigate the effects of interleukin-31（IL-31） or tacrolimus on expression of neurotrophin-4（NT4） in human Hacat cells. Methods：Human keratinocyte cell line（Hacat cells） was cultured and treated with different concentrations of tacrolimus and cell proliferation was determined by the MTT assay. Cultured HaCaT cells were treated with IL-31 alone or in combi－nation with tacrolimus for 24 h. Real-time PCR was used to detect the mRNA of NT4. Levels of NT4 in the culture supernant were measured with ELISA. Results：Tacrolimus with low concentration（10，100） ng/ml had mild effect on cell proliferation（P >0.05），while that with high concentration（1 000 ng/ml） inhibited cell proliferation（P=0.002）. Levels of NT4 in the culture supernant and NT4 mRNA expression were significantly increased in the presence of IL-31. Levels of NT4 in the culture supernant and NT4mRNA expression were significantly decreased with IL-31 and 100 ng/ml tacrolimus acting alone or in combination for 24 h. Conclusion：The expression of NT4 of keratinocyte is up-regulated by IL-31. Tacrolimus might achieve its anti-pruritus effect in atopic dermatitis by decreasing the NT4 expression of keratinocytes.