组胺对皮肤角化上皮屏障功能的作用的研究

Role of histamine in regulation of barrier function in human keratinocytes

目的检测体外培养人皮肤角化上皮细胞在组胺刺激下的跨膜阻力(trans-epithelial resistance,TER)和细胞间通透性的变化,研究组胺对角化上皮屏障功能的作用。方法应用气-液细胞培养技术(air-liquid interface,ALI)培养正常人和特异性皮炎(atopic dermatitis,AD)患者皮肤原代角化上皮细胞,以组胺及其受体拮抗剂等连续刺激上皮细胞96 h,监测TER及细胞间荧光素通过强度,评价皮肤角化上皮屏障功能的改变。结果组胺刺激可以降低皮肤角化上皮细胞的TER,增加上皮细胞间荧光素FITC-葡聚糖(FITC-Dextran)的通透性,作用曲线呈时间和剂量依赖性,组胺刺激与对照组相比差异具有统计学意义;组胺受体1(histamine 1 receptor,H1R)拮抗剂可以抑制组胺对TER的降低作用,而Ca2+载体离子霉素可以诱导类似组胺对TER的降低作用,差异具有统计学意义。结论本实验提示组胺可以降低人皮肤原代角化上皮细胞的屏障功能,有可能与AD等慢性皮肤炎性疾病的发病机制有关,并有可能为临床治疗提供选择依据。

Objective We try to investigate the role of histamine in regulation of barrier function of human keratinocytes by testing the changes of trans-epithelial resistance （ TER ） and paracellular flux of primary human keratinocytes in vitro stimulated by histamine. Methods We cuhured primary human keratinocytes from healthy and atopic dermatitis （AD） individuals in the presence and absence of histamine, histamine receptor blocker for 96 hours in air-liquid interface （ ALI） culture systems. The epithelial barrier function was evaluated by measuring TER and FITC-Dextran paracellular flux. Results Histamine decreased the barrier integrity of human keratinocytes in ALI cultures by impairing TER increase and increasing the paracellular flux to Fitc-Dextran in a dose- and time-dependent manner. There was a statistically significant difference between histamine stimulated group and un-stimulated group. Histamine 1 receptor （H1R） antagonist could block the effect of histamine to TER and Ca^2＋ ionophore ionomyein could induce the similar effect to TER with histamine with statistical significance. Conclusion Our findings that histamine impairs human keratinocytes barrier function might contribute to the pathogenesis of chronic skin inflammatory diseases such as AD and might indicate a possibility for clinical therapeutic strategy.