一氧化氮和白介素-10抑制核因子-κB活化的实验研究

Experimental study of Nitric Oxide and Interleukin-10 inhibiting activation of nuclear factor-κB

目的 探讨一氧化氮 (NO)和白介素 - 10 (IL - 10 )对内毒素 (LPS)诱导的肺泡巨噬细胞核因子 -κB(NF -κB)活化的调节 ,为临床运用提供理论依据。方法 用支气管肺泡灌洗法收集肺泡巨噬细胞 (PAM)进行培养 ,分正常对照组、LPS组、NO +LPS组和IL - 10 +LPS组。用凝胶电泳迁移率改变分析 (EMSA)法和ELISA法分别检测提取物中NF -κB活性和细胞培养上清中TNF -α含量。结果 LPS组NF -κB活性和TNF -α含量在刺激后 0 5～ 4h显著高于正常对照组 (P <0 0 1) ;NO+LPS组和IL - 10 +LPS组的NF -κB活性和TNF -α含量与LPS组相比均明显下降 ,尤在刺激后 1h最显著 (P <0 0 1)。结论 LPS诱导PAM的NF -κB活化 ,导致TNF -α基因表达增强 ;NO和IL - 10可抑制NF -κB活化 ,减少TNF -α的释放 ,缓解LPS诱导的ALI。

Objective To investigate activation of nuclear factor-κB(NF-κB)in lipopoysaccharide-induced pulmonary alveolar macrophages(PAM)stimulated with NO and interleukin-10 Methods PAM collected by BALT were cultured and divided into four groups:control group;LPS-stimuted group;NO+LPS group and IL-10+LPS group The NF-κB activity of nuclear protein extract from the PAM and the concentration of TNF-α in the supernatant were measured by electrophoretic mobility shift assay(ESMA)and ELISA Results The activity of NF-κB and the level of TNF-α significantly increased after LPS stimulation;Compared with LPS-stimulated group,both NF-κB activity and concentration of TNF-α were significantly lowered in NO+LPS group and IL-10+LPS group,especially at 1 hour ( P <0 01) Conclusion LPS might activity NF-κB in PAMs and induce the increase of transcription and expression of TNF-α gene;NO and IL-10 could inhibit the activation of NF-κB and reduce release of TNF-α