摘要
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-α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-α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-κB 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-κBmRNA表达无影响(0.4716±0.0616vs0.4994±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-α,具有调节单核细胞免疫功能的作用,其抑制TNF-α分泌功能可能与其下调单核细胞内NF-κB表达有关。
Objective： To study the role of Novaferon on TNF-a production and expression of TNF-a B mRNA in monocytes isolated from normal human peripheral blood and to provide theoretical basis for treatment of immunological diseases with Novaferon.
Methods： 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-kB mRNA by RT-PCR.
Results： Novaferon inhibited TNF-a 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〈0.01], and the rate was 29.7%. There was no significant change in TNF-a 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-kB mRNA expression in group D compared with group B （0.2829±0.0365 vs 0.4994±0.0604, P〈0.01）. There was no significant change in NF-kB mRNA expression in group C and group E compared with group B （0.4716±0.0616 vs 0.4994±0.0604, P〉0.05; 0.4767±0.0600 vs 0.4994±0.0604, P〉0.05）.
Conclusion： Novaferon can suppress TNF-a 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-kB mRNA expression in monocytes.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2013年第1期66-69,共4页
Journal of Central South University ：Medical Science
