【摘要】 目的探讨NOD样受体热蛋白结构区域3（NOD-like receptor pyrin domain containing 3, NLRP3）炎性小体通路在B族维生素干预孕期PM_2.5致脑损伤中的表达变化,明确B族维生素改善PM_2.5污染所致危害的作用机制。方法建立孕期PM_2.5损伤动物模型,并给予B族维生素干预。将孕鼠随机分为PBS对照组,PM_2.5模型组和VB干预组。分离孕鼠大脑皮层,采用RT-qPCR技术、Western blot技术、ELISA及免疫荧光染色方法检测孕鼠大脑皮层炎性小体通路NLRP3、cleaved caspase-1和IL-1β的表达变化。结果 RT-qPCR结果显示,模型组孕鼠大脑皮层NLRP3、IL-1βmRNA表达水平明显高于对照组;而干预组明显低于模型组,但与对照组无明显差异。Western blot和ELISA检测结果显示,模型组孕鼠大脑皮层NLRP3、cleaved caspase-1和IL-1β的蛋白表达较对照组明显增加,而干预组则明显低于模型组。免疫荧光染色方法显示,干预组孕鼠大脑皮层中NLRP3、cleaved caspase-1免疫反应阳性细胞数较模型组明显减少,与对照组无明显差异。结论孕期给予B族维生素干预可以抑制孕鼠大脑皮层NLRP3炎性小体通路激活,对PM_2.5暴露所致孕鼠大脑皮层损伤可能有一定的保护作用。更多还原

【Abstract】 Objective To investigate the expression changes of proteins in NOD-like receptor pyrin domain containing 3（NLRP3） inflammasome pathway in brain injury induced by PM_2.5 during pregnancy after intervention with B vitamins, and to elucidate the mechanism by which B vitamins improves the brain injury induced by PM_2.5. Methods The animal model of brain injury induced by PM_2.5 during pregnancy was established and intervened with B vitamins. Pregnant mice were randomly divided into the PBS control group, PM_2.5 model group, and VB intervention group. The cerebral cortex of pregnant mice was isolated, and the expression of NLRP3, cleaved caspase-1 and IL-1β in the inflammasome pathway of the cerebral cortex of pregnant mice was detected by RT-qPCR, Western blot, ELISA and immunofluorescence staining. Results RT-q PCR results showed that the expression levels of NLRP3 and IL-1β in the cerebral cortex of pregnant mice in the model group were significantly higher than those in the control group, while those in the intervention group were significantly lower than those in the model group, but there was no significant difference between the intervention group and the control group. Western blot and ELISA results showed that the protein expression of NLRP3, cleaved caspase-1 and IL-1β in the cerebral cortex of pregnant mice in the model group was significantly higher than that in the control group, while that in the intervention group were significantly lower than that in the model group. Immunofluorescence staining showed that both the number of NLRP3 positive cells and the number of cleaved caspase-1 positive cells in the cerebral cortex of pregnant mice in the intervention group were significantly smaller than those in the model group, and there was no significant difference between the intervention group and the control group. Conclusion Intervention with B vitamins during pregnancy inhibits the activation of NLRP3 inflammasome pathway in the cerebral cortex of pregnant mice, which may have a protective effect on the cerebral cortex injury of pregnant mice induced by PM_2.5 exposure.更多还原