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目的探讨叶酸对低氧诱导人单核细胞系THP-1细胞产生炎症介质的影响及对核因子-κB的效应。方法采用不同叶酸浓度(0.4μg/m L,4μg/m L和40μg/m L)预处理细胞30min,再在1%O_2条件下培养细胞24h诱导低氧损伤,收集细胞以及培养上清。通过MTT法检测不同浓度叶酸对细胞活性的影响;ELISA法和实时荧光定量PCR分别检测细胞炎性介质(IL-1β和IL-8)的蛋白和m RNA表达水平;Western blot测定细胞NF-κB p65的蛋白表达。结果0.4~40μg/m L浓度叶酸对THP-1细胞的活性没有影响(P>0.05);叶酸能降低低氧所致THP-1细胞产生炎症介质IL-1β和IL-8蛋白以及m RNA的高表达(P<0.05),同时下调NF-κB p65蛋白表达(P<0.05),并呈现剂量反应关系。结论叶酸能够缓解低氧所引起的细胞炎症反应,其作用可能与抑制THP-1细胞NF-κB通路的激活及其炎症介质的释放相关。
Objective To investigate the effect of folic acid on the inflammatory mediators induced by hypoxia in human monocytic THP-1 cells and its effect on nuclear factor-κB. Methods The cells were pretreated with different concentrations of folic acid (0.4μg / ml, 4μg / ml and 40μg / ml) for 30min. Cells were cultured under 1% O2 for 24 hours to induce hypoxia injury. Cells were harvested and culture supernatants were harvested. The effect of folic acid at different concentrations on cell viability was detected by MTT assay. The protein and m RNA expression of inflammatory mediators (IL-1β and IL-8) were detected by ELISA and real-time fluorescence quantitative PCR respectively. The expressions of NF-κB p65 protein expression. Results Folic acid had no effect on the activity of THP-1 cells in the concentration of 0.4 ~ 40μg / mL (P> 0.05). Folic acid reduced the production of IL-1β, IL-8 and m RNA in THP-1 cells induced by hypoxia (P <0.05), and down-regulated the expression of NF-κB p65 protein (P <0.05), and showed a dose-response relationship. Conclusion Folic acid can alleviate the cellular inflammatory response induced by hypoxia, which may be related to the inhibition of the activation of NF-κB pathway and the release of inflammatory mediators in THP-1 cells.