利多卡因对金黄色葡萄球菌外毒素TSST-1刺激的特应性皮炎患者外周血单一核细胞的影响  

Effects of lidocaine on peripheral blood mononuclear cells from patients with atopic dermatitis stimulated by the Staphylococcus aureus exotoxin TSST-1

作　　者：王媛媛[1] 胡明洁[1] 张静[1] 黄银久[1] 汤必奎[1] 陈昌杰[1] 吴守伟[1] 

机构地区：[1]蚌埠医学院生物科学系,安徽233030

出　　处：《中华皮肤科杂志》2015年第1期28-32,共5页Chinese Journal of Dermatology

基　　金：安徽省高校省级优秀青年人才基金重点项目（2013SQRL050ZD）;蚌埠医学院科技发展基金（Bykf13A01）

摘　　要：目的 探讨利多卡因对金黄色葡萄球菌外毒素激活特应性皮炎患者单一核细胞的影响.方法 抽取6例特应性皮炎患者外周血,常规分离培养单一核细胞.以金黄色葡萄球菌外毒素刺激共孵育,同时加入不同浓度的利多卡因.3H-TdR掺入法检测单一核细胞增殖,酶联免疫吸附法（ELISA）检测Th1和Th2细胞因子的释放.Western印迹法检测利多卡因对与中毒性休克综合征毒素1（TSST-1）刺激的特应性皮炎患者单一核细胞共孵育的HaCaT细胞中间丝相关蛋白的表达.结果 TSST-1（100 μg/L）显著刺激了特应性皮炎患者单一核细胞的增殖（SI=75±2.12,P＜0.05）,并刺激α肿瘤坏死因子、γ干扰素、白细胞介素（IL）2、IL-12、IL-4、IL-5、IL-13细胞因子的释放（均P＜0.05）.CCK-8检测结果显示,与空白对照组相比,100 μmol/L利多卡因显著抑制了TSST-1刺激的特应性皮炎患者单一核细胞的增殖（SI=58±3.14,P＜0.05）,亦显著抑制了TSST-1刺激的特应性皮炎患者外周血IL-4、IL-5、IL-13、α肿瘤坏死因子以及γ干扰素的释放,差异有统计学意义（均P＜ 0.05）.Western印迹法结果显示,100 μmol/L利多卡因阻断了HaCaT细胞中间丝相关蛋白表达的下调,差异有统计学意义（P＜ 0.01）.结论 利多卡因对TSST-1刺激的特应性皮炎患者单一核细胞的激活具有明显的抑制作用.Objective To investigate the effect of lidocaine on Staphylococcus aureus exotoxin-stimulated peripheral blood mononuclear cells （PBMCs） from patients with atopic dermatitis （AD）.Methods Peripheral blood samples were collected from 6 patients with AD,and PBMCs were isolated by a routine method.Then,the PBMCs were stimulated by the Staphylococcus aureus exotoxin toxic shock syndrome toxin-1 （TSST-1） in the absence or presence of lidocaine at varying concentrations.The 3H-TdR incorporation method was performed to detect the proliferation of monocytes,and enzyme-linked immunosorbent assay （ELISA） to quantify the levels of T helper type 1 （Th1） and Th2 cytokines released by PBMCs.Human HaCaT keratinocytes were co-cultured with lidocaine-and TSST-1-stimulated PBMCs from patients with AD for 72 hours,then,Western blot was conducted to examine the expression of filaggrin protein in HaCaT cells.Results TSST-1 （100 μg/L） significantly enhanced the proliferation of PBMCs from patients with AD （stimulation index =75 ± 2.12,P 〈 0.05）,as well as the release of tumor necrosis factor-α （TNF-α）,interferon （IFN）-γ,interleukin （IL）-2,IL-12,IL-4,IL-5 and IL-13 by the PBMCs （all P 〈 0.05）.Compared with the blank control group,100 μmol/L lidocaine significantly inhibited the TSST-1-stimulated proliferation of PBMCs from patients with AD （stimulation index =58 ± 3.14,P〈 0.05）,as well as the release of IL-4,IL-5,IL-13,TNF-α and IFN-γ by the stimulated PBMCs （all P 〈 0.05）.Western blot showed that 100 μmol/L lidocaine significantly blocked the down-regulation of filaggrin expression in HaCaT cells （P 〈 0.01）.Conclusion Lidocaine has a significant inhibitory effect on the activation of TSST-1-stimulated PBMCs from patients with AD.

关 键 词：特应性皮炎 金黄色葡萄球菌 外毒素类 利多卡因 单核细胞 

分 类 号：R758.2[医药卫生—皮肤病学与性病学]