柚皮苷对人皮肤角质细胞分泌趋化因子RANTES及其核转录因子-kB信号通路影响的实验研究

目的：研究炎症性细胞因子肿瘤坏死因子α（TNF-α）和γ干扰素（IFN-γ）对人皮肤角质细胞（HaCaT）分泌趋化因子RANTES的诱导作用；以阿维A为阳性对照药，研究柚皮苷对此诱导作用的抑制作用及其机制。方法：将HacaT分为阴性对照组、模型刺激组（TNF-α＋IFN—γ）、阿维A预处理组（TNF-α＋IFN—γ＋阿维A）和柚皮苷预处理组（TNF-α＋IFN—γ＋柚皮苷）等；采用酶联免疫吸附（ELISA）法测定HaCaT细胞经单用和合用TNF-α、IFN-γ刺激以及加入阿维A和柚皮苷后RANTES的分泌量。采用免疫细胞化学方法和Western blot法检测HaCaT中核转录因子-KB P65（NF—kB P65）蛋白的表达。结果：单用或合用TNF—α、IFN-γ可显著诱导细胞分泌RANTES，并以合用组作用最显著（P〈0．01）。柚皮苷、阿维A均能明显抑制TNF-α、IFN-γ诱导的RANTES的分泌（P〈O．01）及NF—kB P65蛋白的表达。结论：柚皮苷可以抑制TNF-α、INF-γ诱导RANTES的分泌及相关蛋白的产生，其机制可能是通过抑制NF—kB信号传导途径的活化而实现。

Study on the Effect of Naringin on Chemokine RANTES Secretion in Human Keratinocyte and NF-kB Signal Pathway

OBJECTIVE： To study the effect of inflammatory cytokine TNF-α and IFN-γ on R.ANTES secretion in human keratinocyte （HaCaT） and the inhibition effect of naringin on induction mechanism by using acitretin as positive control. METHODS： The HaCaT cells were assigned into negative control group, model stimulation group （treated with TNF-α＋IFN-γ）, acitretin pretreatment group （treated with TNF-α＋IFN-γ＋acitretin）, naringin pretreatment group （treated with TNF-α＋IFN-α,＋naringin）. The concentration of RANTES in HaCaT cells which were stimulated by cytokine TNF-α and IFN-γ alone or together after acitretin and naringin were added into stimulators was determined by ELISA assay. The expressions of NF-κB P65 protein in HaCaT cells was detected with immunocytochemical method and Western blot method. RESULTS： The concenctration of RANTES in HaCaT cells which were stimulated by cytokine TNF-α or IFN-γ alone or together increased; the greatest concentration was observed in model stimulation group （P〈0.01）. Naringin and acitretin limited the effect of cytokine TNF-α and IFN-γ on RANTES and formation of NF-κB protein. CONCLUSION： Naringin can inhibit formation of NF-κB protein and RANTES induced by TNF-α and IFN-γ, which may be achieved by inhibiting the activation of NF-κB signal pathway.