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目的观察梭曼染毒大鼠主要脑区炎性因子IL-1β、TNF-α及核转录调节因子NF-κB在不同时间的表达变化。方法建立大鼠梭曼致惊厥模型,采用HE染色法观察染毒后不同时段动物脑内梨状皮质及海马CA1区病理学改变;采用实时荧光定量PCR(RT-qPCR)及Western印迹测定上述两个脑区IL-1β、TNF-α及NF-κB mRNA和蛋白在不同时间的表达变化。结果病理学结果显示,梭曼中毒所致惊厥大鼠脑内梨状皮质及海马CA1区在中毒后30 min即出现明显病理学损伤,染毒后7 d仍持续存在。RT-qPCR测定结果发现,梨状皮质及海马CA1区内TNF-α、IL-1βmRNA在染毒后30 min表达明显增加,染毒后6 h达峰值,染毒后48 h仍大量表达;NF-κB mRNA表达变化相对较晚,在染毒后6 h出现显著上调,染毒后48 h表达仍异常增高。Western印迹结果表明,海马CA1区TNF-α及IL-1β的蛋白表达总体与mRNA表达类似,但梨状皮质TNF-α及IL-1β的蛋白表达变化总体出现较晚;而NF-κB的蛋白表达在梭曼中毒早期即出现显著升高。结论梭曼中毒大鼠主要脑区内炎性因子IL-1β、TNF-α、NF-κB mRNA及蛋白表达水平在中毒早期出现显著上调,提示针对炎性反应的干预措施可能成为梭曼中毒所致脑损伤的新治疗手段。
Objective To observe the changes of the expression of IL-1β, TNF-α and NF-κB in major brain regions of soman-exposed rats at different time points. Methods The rat model of somatosensory convulsion was established. The pathological changes of piriform cortex and hippocampal CA1 region were observed by HE staining at different time points after injection. The expression levels of these two genes were detected by real-time fluorescence quantitative PCR (RT-qPCR) and Western blot The changes of IL-1β, TNF-αand NF-κB mRNA and protein in different brain regions at different time points were observed. Results The pathological results showed that the pathological lesions of piriform cortex and hippocampal CA1 area of rats with convulsion induced by soman ’s poisoning occurred at 30 min after the poisoning, and persisted after 7 d. The results of RT-qPCR showed that the expression of TNF-α and IL-1βmRNA in piriform cortex and hippocampal CA1 area increased significantly at 30 min and reached the peak at 6 h after exposure, The expression of -κB mRNA was relatively late, and was significantly up-regulated at 6 hours after exposure. The expression of -κB mRNA was still abnormally increased at 48 hours after exposure. Western blot results showed that the protein expression of TNF-α and IL-1β in CA1 area of hippocampus was similar to that of mRNA expression, but the changes of protein expression of piriform cortical TNF-α and IL-1β appeared later in general. The expression of soman early poisoning that appears significant increase. Conclusions The mRNA and protein expressions of IL-1β, TNF-α and NF-κB in the main brain regions of soman rats were significantly increased at the early stage of poisoning, suggesting that the interventions aimed at the inflammatory response may become Soman’s poisoning New treatment for brain damage.