摘要目的 探讨艾司洛尔对内毒素刺激大鼠腹腔巨噬细胞的保护作用及其相关机制.方法 对大鼠腹腔巨噬细胞进行体外培养;随机数字法分四组,分别为正常组、脂多糖(LPS)刺激组、艾司洛尔组、艾司洛尔与脂多糖(LPS)共同刺激组;用qRT-PCR及Western blot分别检测肿瘤坏死因子α(TNF-α)、β1肾上腺素受体(β1-AR)及p65的mRNA及蛋白含量;用EMSA检测各组NF-κB活性.结果 LPS刺激组中TNF-α mRNA与蛋白水平的表达明显高于正常组(P＜0.01),而艾司洛尔与LPS共同刺激组中TNF-α的表达明显低于LPS刺激组(P＜0.01).LPS刺激组中β1-AR的表达水平显著高于正常组(P＜0.01),艾司洛尔组β1-AR的表达水平少于正常组(P ＜0.05),而共同刺激组中β1-AR的表达水平较LPS刺激组减少(P＜0.01).与正常组比,LPS刺激组中p65的表达明显增多(P＜0.01),而共同刺激组中p65的表达明显低于LPS刺激组(P＜0.05).LPS刺激组中NF-κBDNA结合力明显高于正常组(P＜0.01),与LPS刺激组相比,艾司洛尔组与共同刺激组中NF-κBDNA结合力受到显著抑制,差异有统计学意义(P＜0.01).结论 艾司洛尔对LPS诱导大鼠腹腔巨噬细胞中TNF-α表达的减少可能通过对β1受体调控作用NF-κB通路实现.

abstractsObjective To investigate the protective effects of esmolol on peritoneal macrophages in rats with sepsis.Methods Rat peritoneal macrophages were deprived of serum and divided into four groups,namely normal group,LPS stimulated group,esmolol group and LPS plus esmolol group.Quantitative real-time PCR and Western blot were performed to assay the expressions and protein levels of TNF-α,β1-AR and p65.The activation of NF-κB was determined by electrophoretic mobility shift assay.Results The expression of TNF-α mRNA and protein level was significantly increased in LPS-treated group than those in normal group (P ＜ 0.01),and those in LPS plus esmolol group was significantly lower than those in LPS stimulated group (P ＜ 0.01).The expression of β1-AR mRNA and protein level were significantly higher in LPS-treated group than those in normal group (P ＜ 0.01),The expression of β1-AR mRNA and protein level in the esmolol group was lower than those in normal group (P ＜ 0.01),and those in LPS plus esmolol group was lower than those in LPS stimulated group (P ＜ 0.01).Besides,compared with the normal group,the expression of p65 mRNA in the LPS stimulated group was noticeably higher (P ＜ 0.01).The expression of P65 mRNA in LPS plus esmolol group was significantly lower than that in LPS-treated group (P ＜ 0.01).The NF-κB/DNA binding force in the LPS stimulated group was significantly higher than that in normal group (P ＜0.01).Compared with the LPS stimulated group,the NF-κB/DNA binding force in the LPS plus esmolol group was significantly lower (P ＜ 0.01).Conclusions The expression of TNF-α mRNA induced by LPS was decreased by esmolol in rat peritoneal macrophages which might be achieved via inhibiting NF-κB pathway by the modulation role of β1 adrenergic receptor.