利多卡因对脂多糖诱导小鼠巨噬细胞caspase-1和IL-1β表达的影响

Effects of lidocaine on caspase-1 and IL-1β lipopolysaccharide-induced mice macrophages

目的观察利多卡因对脂多糖诱导小鼠巨噬细胞凋亡、含半胱氨酸的天冬氨酸蛋白水解酶-1(caspase-1)和白细胞介素-1β(IL-1β)表达的影响,探讨利多卡因在脂多糖诱导相关炎症保护作用的机制。方法将单层培养于6孔板的小鼠巨噬细胞随机分为对照组、利多卡因组、脂多糖组和利多卡因+脂多糖组。采用流式细胞术测定细胞凋亡率,RT-PCR法检测细胞中caspase-1和IL-1β的mRNA表达水平,ELISA法检测培养液中caspase-1和IL-1β的蛋白浓度。结果与对照组相比,利多卡因组细胞凋亡率、caspase-1和IL-1β的mRNA表达及蛋白表达无明显变化(P>0.05),脂多糖组细胞凋亡率显著增高(P<0.05),caspase-1和IL-1β的mRNA表达水平以及蛋白表达水平显著增高(P<0.05);利多卡因+脂多糖组与脂多糖组相比细胞凋亡率显著降低(P<0.05),caspase-1和IL-1β的mRNA表达水平以及蛋白表达水平显著降低(P<0.05)。结论利多卡因预处理可抑制脂多糖诱导小鼠巨噬细胞的凋亡和caspase-1、IL-1β过表达。利多卡因抗炎作用可能与抑制免疫细胞凋亡、减少炎性因子释放有关。

Objective To observe the protective effect of lidocaine on cysteinyl aspartate specific proteinase-1（ caspase-1） and interleukin-1β（ IL-1β） in lipopolysaccharide-induced inflammation in mice and to investigate its mechanism.Methods Monolayer cultured mouse macrophages in six-well plates were randomly divided into 4 groups： control group,lidocaine group,lipopolysaccharide group,and lidocaine ＋ lipopolysaccharide group. Cell apoptosis rate was measured with flowcytometry. Expressions of caspase-1 and IL-1β mRNA were detected with realtime PCR（ RT-PCR）. The protein concentrations of caspase-1 and IL-1β in the culture medium were assessed with enzyme-linked immunosorbent assay（ ELISA）. Results Compared with control group,in lidocaine group,the apoptosis rate,mRNA expression and protein expression of caspase-1 and IL-1β had no significant change（ P〉0. 05）; in the lipopolysaccharide group,the apoptosis rate,mRNA expression and protein expression of caspase-1 and IL-1β were significantly higher（ all P〈0. 05）. Compared with the lipopolysaccharide group,in lidocaine ＋ lipopolysaccharide group,apoptosis was significantly reduced（ P〈0. 05）,mRNA and protein expression levels of caspase-1 and IL-1β expressions were markedly lower（ P〈0. 05）. Conclusion Lidocaine can reduce apoptosis in murine macrophages lipopolysaccharideinduced inflammatory response. Protective effects of lidocaine on tissue inflammation may be related to inhibition ofcaspase-1 and IL-1β.