高压氧对急性有机磷中毒大鼠脑保护作用的实验研究

发布时间：2018-08-07 21:19

【摘要】：目的：通过建立大鼠急性有机磷中毒（AOPP）模型，研究缺氧诱导因子-1(HIF-1)在脑组织中的表达和血清中丙二醛（MDA）含量的变化，，以及高压氧对它们的干预作用，探讨高压氧防治急性有机磷中毒性脑损伤的作用及其机制，为拓展高压氧治疗的范围提供理论依据。方法：健康雄性SD大鼠60只，随机分为正常对照组、中毒组、常规治疗组和高压氧治疗组，正常对照组6只，其余3组各18只。中毒组、常规治疗组和高压氧治疗组分3个时间点（建模后1，3，7h）进行标本采集，每个时间点各6只。采用颈背部皮下累计注射法造模，每次敌敌畏用量4mg/kg，共注射5次，保证大鼠不间断抽搐3小时。常规治疗组在造模后立即予长托宁0.12mg/kg、氯解磷定30mg/kg大腿外侧肌肉注射，高压氧治疗组在常规治疗之后即行高压氧治疗，正常对照组注射等容量的生理盐水。各组在造模完成后1、3、7h处死大鼠，下腔静脉取血，离心分离血清，采用硫代巴比妥酸比色法试剂盒检测MDA的含量；断头取脑，大脑皮层放入4％的多聚甲醛中固定，石蜡包埋、切片，HE染色、尼氏染色观察脑组织病理改变及IHC检测HIF-1蛋白表达；海马组织用于荧光定量PCR检测HIF-1mRNA的表达。所得数据用SPSS19.0软件统计分析。结果：与中毒组相比，高压氧治疗组的脑组织病理损伤有所减轻。中毒组各时间点，脑组织HIF-1的表达和血清MDA含量都依次增高，显著高于正常对照组（P0.05）；经高压氧治疗后，脑组织HIF-1的表达和血清MDA含量逐渐下降（P0.05），也低于常规治疗组1h、3h（P0.05）。相关分析发现，HIF-1mRNA相对表达量与血清MDA含量（r=0.909，P=0.000）具有显著的正相关性。结论：高压氧对急性有机磷中毒性脑损伤的保护作用优于常规治疗组，其作用机制与抗氧化损伤和抑制HIF-1的表达有关；且高压氧干预越早越好。
[Abstract]:Objective: to establish (AOPP) model of acute organophosphorus poisoning in rats, to study the expression of hypoxia inducible factor 1 (HIF-1) in brain tissue and the change of malondialdehyde (MDA) content in serum, and the effect of hyperbaric oxygen on it. To investigate the effect of hyperbaric oxygen on acute organophosphorus poisoning brain injury and its mechanism, and to provide theoretical basis for expanding the range of hyperbaric oxygen therapy. Methods: sixty male Sprague-Dawley rats were randomly divided into normal control group, poisoning group, routine treatment group and hyperbaric oxygen treatment group, normal control group (n = 6), and other 3 groups (n = 18 each). Poisoning group, routine treatment group and hyperbaric oxygen treatment group were collected at three time points (1: 3 hours after modeling), 6 rats in each time point. The model was made by subcutaneous injection of dichlorvos at a dose of 4 mg / kg for 5 times in order to ensure continuous convulsion in rats for 3 hours. The routine treatment group was treated with Changtonin 0.12 mg / kg immediately after the model was established, and 30mg/kg was injected intramuscularly on the lateral side of thigh. The hyperbaric oxygen group was treated with hyperbaric oxygen immediately after routine treatment, and the normal control group was injected with normal saline of the same volume. The rats in each group were killed at 1: 3h after modeling, blood was collected from inferior vena cava, serum was isolated by centrifugation, the content of MDA was detected by thiobarbituric acid colorimetric kit, the brain was taken off the head, the cerebral cortex was fixed in 4% polyformaldehyde, and embedded in paraffin. The pathological changes of brain tissue and the expression of HIF-1 protein were detected by IHC, and the expression of HIF-1mRNA was detected by fluorescence quantitative PCR in hippocampal tissue. The data were analyzed by SPSS19.0 software. Results: compared with the poisoning group, hyperbaric oxygen treatment group reduced the pathological injury of brain tissue. At each time point, the expression of HIF-1 and the content of serum MDA in the poisoning group increased in turn, which were significantly higher than those in the normal control group (P0.05); after hyperbaric oxygen treatment, the expression of HIF-1 and the content of serum MDA decreased gradually (P0.05), and were also lower than those in the routine treatment group for 1 hour and 3 hours (P0.05). Correlation analysis showed that there was a significant positive correlation between the relative expression of HIF-1 mRNA and the level of serum MDA (r 0.909 Pu 0.000). Conclusion: the protective effect of hyperbaric oxygen on acute organophosphorus poisoning brain injury is better than that of routine treatment group, and its mechanism is related to antioxidant damage and inhibition of HIF-1 expression, and hyperbaric oxygen intervention is better as early as possible.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R595.4

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