不同时间脓毒症核转录因子-κB对细胞因子表达调节的实验研究

Correlation between nuclear transcription factor (NF-κB) and cytokines at 4hs or 18hs in mice with sepsis

目的探讨在不同时间段内脓毒症核转录因子-κB(NuclearTranscriptionFactor,NF-κB)对细胞因子表达的调节及其临床意义。方法将30只小鼠分为正常对照组、假手术(sham)组和盲肠结扎(CLP)组,采用酶联免疫吸附法(ELISA)和RT-PCR法分别检测血清细胞因子(TNFα、IL-6和IL-10)总量和脾细胞相应细胞因子基因表达量。同时采用电泳迁移率滞后分析法检测核转录因子NF-κBDNA结合活性。结果CLP组细胞因子TNFα、IL-6和IL-10高于正常对照组和sham组;CLP组有活性的NF-κB增多,以正调节促炎细胞因子TNFα和IL-6表达为主,TNFα和IL-6基因表达增高,且在4～18h调节IL-6(P<0.01)的作用高于TNFα(P<0.05),而对抗炎因子IL-10的强化调节作用略有下降(P>0.05)。结论随着脓毒症时间的推移,活性NF-κB增多,对促炎因子的正调节作用高于抗炎因子,是造成促炎因子与抗炎因子不平衡的重要原因。不同时间脓毒症细胞因子失衡程度不同,脓毒症的发展趋势亦不同。

[ Objective] To investigate correlation between Nuclear Transcription Factor（NF-κB） and cytokines at 4 hs or 18 hs in mice with CLP. [Methods] 30 C57BL/6 mice were randomly divided into normal control group, sham group and CLP group. ELISA was used to measure the levels of TNFα, IL-6 and IL-10 in sera. RNA from the spleen cells was isolated to determine the expression of TNFα, IL-6 and IL-10 mRNA with reverse transcription polymerase chain reaction. The activity of NF-κB was measured by using electrophresis mobility assay. [Results] The levels of TNFα, IL-6 and IL-10 are significantly higher in mice with CLP than in those of the normal and sham controls; Activity of NF-κB is increased in CLP mice after both 4hs and 18 hs. TNFα and IL-6 were up-regulated by active NF-κB at 4 hs following septic challenge （P 〈0.05）. Strong positive correlations existed between activity of NF-κB and expression of IL-6 （P 〈0.05）, while IL-10 levels shows a tendency to slightly decrease （P 〉0.05）. [Conclusions] More hours in mice with CLP, more active NF-κB is produced. The balance between proinflammatory and anti-inflammatory cytokines is lost with increasing the quantity of NF-κB. The deteriorism of septic challenge is decided by the severity to unbalance.