Objective: To study the role of Novaferon on TNF-a production and expression of NF-ΚB mRNA in monocytes isolated from normal human peripheral blood and to provide theoretical basis for treatment of immunological diseases with Novaferon.rnMethods: Monocytes were isolated from the peripheral blood in 30 healthy volunteers and divided into 5 groups: group A was blank control, group B was stimulated by LPS without Novaferon intervention, group C by LPS together with Novaferon intervention, group D by LPS before Novaferon intervention, which group E by LPS after Novaferon intervention. We detected the concentration of TNF-a after LPS stimulation and Novaferon intervention in the supernatant by ELISA and expression of NF-ΚB mRNA by RT-PCR.rnResults: Novaferon inhibited TNF-α production by monocytes isolated from healthy volunteers induced by LPS in vitro in group D compared with group B [(1446.76±72.36) pg/mL vs (946.46±46.12) pg/mL, P<0.0l], and the rate was 29.7%. There was no significant change in TNF-α concentration in group C and E compared with group B [(1446.76+72.36) pg/mL vs (1275.62±87.75) pg/mL, P>0.05; (1446.76+72.36) pg/mL vs (1383.62+86.96) pg/mL, P>0.05]. There was significant change in NF-ΚB mRNA expression in group D compared with group B (0.2829±0.0365 vs 0.4994±0.0604, P<0.0l). There was no significant change in NF-ΚB mRNA expression in group C and group E compared with group B (0.47l6±0.0616 vs 0.4994±0.0604, P>0.05; 0.4767±0.0600 vs 0.4994±0.0604, P>0.05).rnConclusion: Novaferon can suppress TNF-α secretion by monocytes induced by LPS in vitro, and it can affect the immunity function of monocytes, which may be associated with the downregulation of NF-ΚB mRNA expression in monocytes.%目的:探讨乐复能在体外对LPS介导的健康人外周血单核细胞分泌TNF-a及NF-κB mRNA表达的影响,以期为乐复能治疗克罗恩病等免疫性疾病提供理论依据.方法:分离30例健康人外周血单核细胞并进行体外培养,分别按以下5种方法(5组)进行体外实验:A组为空白对照组；B组为单纯LPS刺激组；C组为LPS与乐复能同时加入组；D组为先加入LPS刺激,后加入乐复能组；E组为先加入乐复能,后加入LPS刺激组.干预后用ELISA法检测培养液内TNF-α浓度,然后采用RT-PCR方法检测单核细胞内NF-κB mRNA表达情况.结果:基础状态下,体外培养的健康人单核细胞分泌少量TNF-α[(470.23±35.24) pg/mL)],加入LPS刺激后,TNF-a的分泌明显增加[(1446.76±72.36) pg/mL)],在LPS刺激后再加入乐复能,TNF-α的分泌明显减少[(1446.76±72.36) pg/mL vs (946.46±46.12)pg/mL,P＜0.01],下降约29.7％.而乐复能在LPS刺激前或与LPS同时加入培养细胞内时,对TNF-α分泌无影响[(1446.76±72.36) pg/mL vs (1275.62±87.75) pg/mL,P＞0.05; (1446.76±72.36) pg/mL vs (1383.62±86.96) pg/mL,P＞0.05].乐复能明显下调经LPS诱导的单核细胞内NF-1B mRNA表达(0.2829±0.0365 vs 0.4994±0.0604,P＜0.01),而对于未提前接受LPS刺激的单核细胞,乐复能对其NF-κB mRNA表达无影响(0.4716±0.0616 vs 0.4994±0.0604,P＞0.05; 0.4767±0.0600vs 0.4994±0.0604,P＞0.05).结论:乐复能在体外能抑制LPS介导的健康人外周血单核细胞分泌TNF-a,具有调节单核细胞免疫功能的作用,其抑制TNF-a分泌功能可能与其下调单核细胞内NF-κB表达有关.
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