利多卡因对肿瘤坏死因子α诱导中性粒细胞凋亡的影响(英文)

Effects of lidocaine on neutrophil apoptosis induced by tumor necrosis factor alpha

背景:利多卡因被认为有抗炎作用,而其作用机制不清。核转录因子κB是炎症反应的关键环节。目的:观察利多卡因对人体中性粒细胞核转录因子κB激活和中性粒细胞凋亡的影响。设计、时间及地点:对比观察,于2006-10/2007-02在江苏省麻醉学重点实验室完成。材料:肿瘤坏死因子α(O127:B8)购于Sigma公司;外周血标本健康者,受试者知情同意。方法:人体中性粒细胞分为5组:生理盐水对照组、肿瘤坏死因子α组、肿瘤坏死因子α+利多卡因1.0mmol/L组、肿瘤坏死因子α+利多卡因2.0mmol/L组、肿瘤坏死因子α+利多卡因4.0mmol/L组,共同孵育3h。主要观察指标:以反转录聚合酶链反应及免疫印迹法检测利多卡因对核转录因子κBmRNA和I-κBmRNA表达及蛋白含量的影响。孵育12h和24h,以流式细胞术分析对中性粒细胞凋亡的影响。结果:利多卡因组核转录因子κBmRNA表达显著降低,而I-κBmRNA表达显著增加。并且利多卡因2.0mmol/L组和4.0mmol/L组显著优于1.0mmol/L组(P<0.05),而2.0mmol/L组和4.0mmol/L组之间差异不显著(P>0.05)。利多卡因在12h和24h都可以显著抑制肿瘤坏死因子α诱导的外周血中性粒细胞凋亡(P<0.05),而4.0mmol/L组显著优于1.0mmol/L组(P<0.05)。结论:利多卡因1.0,2.0,4.0mmol/L都可以显著下调肿瘤坏死因子α诱导的外周血中性粒细胞P65mRNA的表达,并且在12和24h可以部分逆转肿瘤坏死因子α诱导的中性粒细胞凋亡。

BACKGROUND： Lidocaine is deemed to can attenuate the inflammatory response, but the mechanisms are poorly understood. Nuclear factor- κB （NF- κB） is a key process of imflammatory response. OBJECTIVE： To determine the effect of tidocaine on activation and apoptosis of NF-κ B in human neutrophils in vitro. DESIGN, TIME AND SETTING： The controlled experiment was conducted at the Jiangsu Provincial Key Laboratory of Anesthesiology from October 2006 to February 2007. MATERIALS： Tumor necrosis factor （TNF）-α （0127：B8） was purchased from Sigma, USA. Peripheral blood samples were obtained from healthy donors after their informed consent. METHODS： Human neutrophilic granulocytes were assigned into five groups： （1） saline control, （2） TNF- α, （3） TNF-α ＋ lidocaine 1.0 mmol/L, （4）TNF- α ＋ lidocaine 2.0 mmol/L, and （5） TNF- α ＋ lidocaine 4.0 mmol/L groups. Incubation was performed for 3 hours. MAIN OUTCOME MEASURES： The effects of lidocaine on expression of NF- κB mRNA and I- κB mRNA in the cytosol extracts were analyzed by reverse transcription-polymerase chain reaction （RT-PCR）, and content of p65 protein were analyzed by Western blot. Neutrophilic granulocyte apoptosis was detected on flow cytometry after incubation 12 hours and 24 hours. RESULTS： The expression of NF- κB mRNA in the nuclear extracts was significantly decreased and I- κB mRNA in the cytosol extracts was markedly increased in the lidocaine group, The expression of NF-κB was significantly better in the 2.0 mmol/L and 4.0 mmol/L lidocaine groups than in the 1.0 mmol/L lidocaine group （P 〈 0.05）. No significant difference was detected between the 2.0 mmol/L lidocaine group and the 4.0 mmol/L lidocaine group （P 〉 0.05）. Lidocaine could significantly inhibit peripheral blood neutrophilic granulocyte apoptosis induced by TNF- α （p 〈 0.05）. The inhibitory effect was significantly better in the 4.0 mmol/L lidocaine group than in the 1.0 mmol/L lidocaine group （P 〈 0.05）. CONCLUSION： Lidocaine （1.0, 2.0 and 4.0 mmol/L） can down-regulate the expressions of NF-κB subunit p65 mRNA and the content of p65 protein in human polymorphoneclear neutrophils, and can significant reverse the reduction of apoptosis induced by TNF- α.