孕期PM2.5暴露影响子代大脑发育中HMGB1/NF-κB通路的作用及机制研究

摘要

目的探讨孕期PM2.5暴露影响子代大脑发育中HMGB1/NF-κB p65通路的作用.方法40只昆明小鼠孕鼠随机分5组,每组8只.气管滴注法向对照组注30 μL PBS,PM2.5低、中、高剂量组各注0.25920、1.56695、3.45600 μg/ μL PM2.5混悬液30 μL,孕期每3天注1次,共7次.空白组无处理.ELISA法检测子代大脑皮层肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)、高迁移率族蛋白1(HMGB1)、核因子κB p65(NF-κB p65)浓度;Westernblot 、RT-PCR法检测HMGB1、NF-κB p65蛋白及mRNA变化.结果ELISA示与对照组相比,高、中剂量组IFN-γ、NF-κB p65升高而HMGB1下降(P＜0.05),高剂量组TNF-α升高(P＜0.05).Western blot示高、中剂量组与对照组相比,HMGB1下降且NF-κB p65上升(P＜0.05).PCR示HMGB1 mRNA无变化,高剂量组NF-κB p65 mRNA高于对照组(P＜0.05).结论孕期PM25暴露使子代鼠大脑皮层产生炎症,HMGB 1/NF-κB通路可能起重要作用.%Objective To investigate the mechanism of HMGB1/NF-κB signal pathway affecting cerebral development of offspring after exposure to PM2.5 during pregnancy.Methods Fourty pregnant Kunming mice were randomly divided into 5 groups,8 rats in each group.PM2.5 low-dose group,medium-dose group and high-dose group were instilled 30 μL PM2.5 suspension of 0.25920,1.56695,3.45600 μg/μL respectively by improved technique for rapid intratracheal instillation.The control group was instilled PBS of the same volume.The blank group received no treatments.Mice were instilled tracheally once every 3 days for 7 times during pregnancy.ELISA was used to measure the levels of TNF-α,IFN-γ,HMGB1 and NF-κB p65 in the cerebral cortex of offspring mice.Western blot and RT-PCR were used to detect the protein and mRNA expression of HMGB1 and NF-κB p65.Results ELISA showed that compared with the control group,the levels ofIFN-γ,NF-κB p65 and HMGB1 in the high and middle dose groups were significantly lower (P ＜ 0.05);high-dose group TNF-α increased (P ＜ 0.05).Western blot showed that HMGB1 decreased and NF-κB p65 increased in the high and middle dose groups compared with the control group (P ＜ 0.05).PCR showed HMGB1 mRNA unchanged;high-dose group NF-κB p65 mRNA was higher than the control group (P ＜ 0.05).Conclusion PM2.5 exposure during pregnancy may cause inflammation in the cerebral cortex of offspring mice,and HMGB1/NF-κB pathway may play an important role.