参附注射液对脂多糖诱导的大鼠肺泡巨噬细胞核因子-κB的激活和细胞因子产生的影响  被引量:6

Effects of Shen-fu Injection on Lipopolysaccharide-induced NF-κB Activity and Cytokine Release in Rat Alveolar Macrophages

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作  者:王进[1] 乔礼芬[2] 李永胜[1] 杨光田[1] 

机构地区:[1]华中科技大学同济医学院附属同济医院急诊科,武汉430030 [2]华中科技大学同济医学院附属同济医院呼吸科,武汉430030

出  处:《华中科技大学学报(医学版)》2009年第1期15-18,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基  金:湖北省科技攻关计划资助项目(No.2003AA301C51)

摘  要:目的研究脂多糖(LPS)诱导的大鼠肺泡巨噬细胞核因子-κB(NF-κB)的激活和细胞因子的释放以及参附注射液(SF)的干预作用,进一步探讨SF对肺脏保护机制。方法通过支气管肺泡灌洗获取大鼠肺泡巨噬细胞(AMs)。对获取的AMs进行LPS刺激(10 ng/ml,2 h)或预先用SF(5μl/ml,10μl/ml)孵育30 min,然后加入LPS(10 ng/ml)分别刺激2 h。用RT-PCR法检测AMs中肿瘤坏死因子-α(TNF-α)的基因表达水平。ELISA法检测培养上清中TNF-α和IL-8的水平。Western blot法检测AMs中NF-κB抑制蛋白-α(IκBα)和NF-κB的水平。结果LPS能够增加AMs中TNF-αmRNA表达水平和培养上清中TNF-α和IL-8的水平。同时LPS促进了IκBα的降解,诱导了NF-κB的激活。与LPS组比较,SF能够减少AMs中TNF-αmRNA表达水平和培养上清中TNF-α和IL-8的水平;抑制LPS诱导的IκBα的降解和NF-κB的激活。结论SF通过抑制AMs中IκBα的降解,减少了NF-κB的激活,从而减少了LPS诱导的大鼠AMs细胞因子的产生。Objective To study the effects of Shen-fu injection (SF) on lipopolysaccharide (LPS)-induced nuclear factor kappa B (NF-κB) activation and cytokine production in rat alveolar macrophages (AMs). Methods AMs were collected from bronchoalveolar lavage fluid from rats. The AMs were incubated with SF (5 and 10 μl/ml) for 30 min, and then stimulated for 2 h by LPS. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression levels of tumor necrosis factor (TNF)-α mRNA. Enzyme-linked immunosorbent assay was used to detect the protein levels of TNF-α and IL-8. Western blot was used to detect the levels of IκBα and NF-κB. Results LPS increased TNF-α mRNA expression, TNF-α and IL-8 levels in the supernatants, and promoted the degradation of IκBα and NF-κB activation. As compared with LPS group, SF decreased TNF-α mRNA expression, TNF-α and IL-8 release, and inhibited IκBα degradation and NF-κB activation. Conclusion SF could attenuate LPS-induced cytokine production in rat AMs by inhibiting IκBα degradation and NF-κB activation.

关 键 词:核因子-ΚB 参附注射液 细胞因子 肺泡巨噬细胞 

分 类 号:R285.5[医药卫生—中药学] Q786[医药卫生—中医学]

 

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