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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-κBP6(5NF-κBP65)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<0.01)及NF-κBP65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-κB信号传导途径的活化而实现。
Objective: To investigate the induction of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) on the secretion of chemokine RANTES by human skin keratinocytes (HaCaT) , To study the inhibitory effect of naringin on this induction and its mechanism. Methods: HaCaT was divided into three groups: negative control group, model stimulus group (TNF-α + IFN-γ), Avia A pretreatment group (TNF-α + IFN-γ + Avia A) and naringin pretreatment group TNF-α + IFN-γ + naringin); HaCaT cells were treated with TNF-α and IFN-γ alone or combined with Avia A and Naringin after enzyme-linked immunosorbent assay (ELISA) The amount of secretion. Immunocytochemistry and Western blot were used to detect the expression of nuclear transcription factor-κBP6 (5NF-κBP65) in HaCaT cells. Results: RANTES was significantly induced by IFN-γ or IFN-γ alone or in combination, and the combination of TNF-α and IFN-γ was the most significant (P <0.01). Naringin and Avia A significantly inhibited TNF-α and IFN-γ-induced RANTES secretion (P <0.01) and NF-κB P65 protein expression. CONCLUSION: Naringin can inhibit TNF-α and INF-γ-induced secretion of RANTES and the production of related proteins, and its mechanism may be through inhibiting the activation of NF-κB signaling pathway.