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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 the regulation of cytokine expression by nuclear transcription factor-κB (NF-κB) in sepsis and its clinical significance at different time points. Methods Thirty mice were divided into normal control group, sham operation group and cecal ligation (CLP) group. Serum cytokines (TNFα, IL- 6 and IL-10) and the corresponding cytokine gene expression of spleen cells. At the same time, the nuclear transcription factor NF-κB DNA binding activity was detected by electrophoretic mobility shift assay. Results The levels of cytokines TNFα, IL-6 and IL-10 in CLP group were higher than those in normal control group and sham group. The activity of NF-κB in CLP group was increased, positively regulated the expression of TNFα and IL-6, (P <0.01) was higher than that of TNFα (P <0.05), while the anti-inflammatory factor IL-10 had a slight decrease (P > 0.05). Conclusion With the passage of sepsis, the activity of NF-κB increased, the positive regulation of proinflammatory cytokines is higher than that of anti-inflammatory cytokines, which is an important reason for the imbalance of proinflammatory cytokines and anti-inflammatory cytokines. Different degrees of sepsis cytokines imbalance in the development trend of sepsis are also different.