铁代谢紊乱在七氟醚和异氟醚致小鼠认知功能障碍中的作用

发布时间：2018-01-04 12:28

本文关键词：铁代谢紊乱在七氟醚和异氟醚致小鼠认知功能障碍中的作用　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本研究拟评价铁代谢紊乱在七氟醚和异氟醚致小鼠认知功能障碍中的作用。方法:清洁级健康雄性昆明小鼠24只,12月龄,体重45~55 g,采用随机数字表法,将其分为3组(n=8):对照组(Con组)、七氟醚组(Sev组)和异氟醚组(Iso组)。Sev组吸入2%七氟醚6 h,Iso组吸入1.4%异氟醚6 h,载气为纯氧,Con组仅吸入纯氧6 h。各组小鼠于麻醉处理前进行Morris水迷宫定位航行训练,历时5 d,每天4次。麻醉结束后12 h再次进行Morris水迷宫实验,通过摄像机和计算机记录各组小鼠水迷宫运动轨迹,逃避潜伏期和寻找平台的总路程。随后,腹腔注射0.4%戊巴比妥钠(1 ml/100 g)麻醉小鼠,剪开胸腔暴露心脏,使用0.9%氯化钠对小鼠进行血液灌流,剪下头部,剥取脑组织,分离出大脑皮层和海马组织。采用Western blot法定性检测各组小鼠皮层和海马区铁代谢相关蛋白ferritin-L、ferritin-H、TfR1、FPN1、DMT1(+IRE)以及磷酸化Tau蛋白表达水平,通过化学发光凝胶成像仪获取相应的蛋白条带图,然后使用Image-Pro Plus 6.0软件分析各蛋白条带的积分光密度值。以目的条带积分光密度值与内参蛋白条带积分光密度值的比值反映目的蛋白的表达水平。采用SPSS13.0软件进行统计分析,以上正态分布的计量资料以均数±标准差(？)表示,组间比较采用单因素方差分析,P0.05认为差异有统计学意义。结果:1 Morris水迷宫实验结果:麻醉处理前,三组小鼠寻找平台的总路程和逃避潜伏期差异无统计学意义(P0.05);与Con组小鼠相比,Sev组和Iso组小鼠寻找平台的总路程和逃避潜伏期均延长(P0.05)。2铁代谢相关蛋白表达水平的检测结果:与Con组相比,Sev组小鼠的大脑皮层及海马中ferritin-L、ferritin-H、FPN1表达均上调,TfR1表达下调(P0.05),DMT1(+IRE)表达差异无统计学意义(P0.05);与Con组相比,Iso组小鼠大脑皮层和海马中ferritin-L、ferritin-H、TfR1、FPN1和DMT1(+IRE)的表达水平差异无统计学意义(P0.05)。3磷酸化Tau蛋白表达水平的检测结果:与Con组相比,Sev组和Iso组小鼠大脑皮层和海马中磷酸化Tau蛋白表达均上调(P0.05)。结论:七氟醚导致小鼠认知功能障碍的机制与铁代谢紊乱有关,而异氟醚导致小鼠认知功能障碍的过程中并没有铁代谢异常的发生。
[Abstract]:Objective: to evaluate the effect of iron metabolism disorder on cognitive dysfunction induced by sevoflurane and isoflurane in mice. The rats were randomly divided into 3 groups: control group (Con group), sevoflurane group (group Sev) and isoflurane group (Iso group) and isoflurane group (group Iso) inhaled sevoflurane for 6 h. The Iso group inhaled 1.4% isoflurane for 6 hours, the carrier gas was pure oxygen for 6 hours only. The mice in each group were trained in Morris water maze navigation for 5 days before anesthesia. Morris water labyrinth experiment was conducted again 12 hours after anesthesia, and the motion track of water maze was recorded by camera and computer. Then, 0.4% pentobarbital sodium (1 ml/100 g) was injected intraperitoneally to anesthetize the mice and cut open the chest cavity to expose the heart. The mice were perfused with 0.9% sodium chloride, the head was cut off and the brain tissue was removed. The cortical and hippocampal tissues were isolated, and the iron metabolism-related protein ferritin-L in cortex and hippocampus of each group was detected qualitatively by Western blot assay. The expression level of DMT1 (IRE1) and phosphorylated Tau protein were obtained by chemiluminescent gel imager. Then use Image-Pro Plus. The integrated optical density of each protein band was analyzed by software 6.0. The expression level of the target protein was reflected by the ratio of the integrated optical density value of the target band to the integral optical density value of the protein band of the internal reference protein. The expression level of the target protein was reflected by SPSS13.0 soft. Statistical analysis was carried out. The measurement data of the above normal distribution are calculated as mean 卤standard deviation? The results showed that the differences were statistically significant by univariate ANOVA (P0.05). Results: 1 Morris water maze experiment results: before anesthesia treatment. There was no significant difference in the total distance and escape latency between the three groups. Compared with Con group mice. The expression of P0.052iron metabolism-related protein in Sev group and Iso group was significantly longer than that in Con group. The results were as follows: compared with Con group, the total distance and escape latency of mice in Sev group and Iso group were prolonged. In Sev group, the expression of ferritin-Lnferritin-Hfritin-HFPN1 was up-regulated in cerebral cortex and hippocampus of mice, and the expression of TfR1 was down-regulated (P0.05). There was no significant difference in the expression of DMT1 (IRE1). Compared with Con group, ferritin-Lnferritin-HfR1 was found in cerebral cortex and hippocampus of mice in Iso group. There was no significant difference between FPN1 and DMT1 (IREs) in the expression of P0.05.3 phosphorylated Tau protein. The results were as follows: compared with Con group. The expression of phosphorylated Tau protein in cerebral cortex and hippocampus of Sev and Iso mice were up-regulated. Conclusion: the mechanism of cognitive dysfunction induced by sevoflurane is related to iron metabolism disorder. There was no abnormal iron metabolism during the cognitive impairment induced by isoflurane in mice.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614

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