七氟醚对小鼠海马区BDNF-TrkB-CREB信号通路表达影响及其氧化应激机制研究
Effects of sevoflurane on the expression of BDNF-TrkB-CREB signaling pathway in mouse hippocampus and its mechanism of oxidative stress
苏凤龙 1刘军 2郭艳苓 3杨颖 3韩玉琴1
作者信息
- 1. 河北省承德市妇幼保健院麻醉科,河北承德 067000
- 2. 河北省承德市妇幼保健院院办,河北承德 067000
- 3. 河北省承德市妇幼保健院产科,河北承德 067000
- 折叠
摘要
目的 从动物实验角度分析七氟醚对小鼠海马区BDNF-TrkB-CREB信号通路表达影响,并探讨其氧化应激机制.方法 以32只6～7周龄SPF级健康雄性C57BL/6小鼠为研究对象,遵循随机化原则及等额原则分为对照组/实验组,每组16只.对照组予生理盐水,腹腔注射,连续注射5 d;采用颈椎脱臼法将小鼠处死.实验组行七氟醚吸入诱导,七氟醚挥发罐开至8％,氧流量6 L/min,监测七氟醚的呼出浓度;待小鼠麻醉成功后采用颈椎脱臼法将小鼠处死.取固定后的小鼠海马组织,依次放入梯度酒精溶液逐步脱去组织水分,再使用二甲苯处理至组织透明.采用ELISA法测定小鼠海马组织氧化应激指标(MDA/GSH-Px/SOD)水平;采用RT-PCR检测BDNF/TrKb/CREB mR-NA表达水平;采用Western Blot检测BDNF/TrKb/CREB蛋白表达水平.结果 与对照组相比,实验组小鼠海马组织中 BDNF mRNA、Trkb mRNA、CREB mRNA 基因表达量显著升高(P＜0.05).实验组 BDNF、Trkb、CREB 蛋白表达水平均显著高于对照组(P＜0.05).与对照组比,实验组小鼠海马组织中MDA水平降低,GSH-Px和SOD水平升高(P＜0.05).对照组小鼠海马组织结构破坏严重,可见大量变性和坏死的神经细胞,且有大量炎性细胞浸润;实验组小鼠海马组织结构完整,神经细胞排列整齐有序,血管丰富,未见炎性细胞浸润、水肿等现象.结论 七氟醚能够上调小鼠海马组织中BDNF mRNA、Trkb mRNA、CREB mRNA水平,而该机制可能与小鼠海马组织抗氧化能力的提升以及保护小鼠海马组织结构不受损伤有关.
Abstract
Objective To analyze the effect of sevoflurane on the expression of BDNF-TrkB-CREB signaling path-way in the hippocampus of mice from the perspective of animal experiments,and to explore its oxidative stress mecha-nism.Methods Thirty-two SPF healthy male C57BL/6 mice aged 6～7 weeks were selected as the research objects,and were divided into control group/experimental group according to the principle of randomization and equal amount,with 16 mice in each group.The control group was given normal saline,intraperitoneally injected,and injected continuously for 5 days;the mice were sacrificed by cervical dislocation.The experimental group was induced by sevoflurane inhala-tion,the sevoflurane vaporizer was opened to 8％,the oxygen flow was 6 L/min,and the exhaled concentration of sevoflurane was monitored;after the mice were successfully anesthetized,the mice were sacrificed by cervical disloca-tion.The fixed mouse hippocampus tissue was taken,successively put into gradient alcohol solution to gradually remove the tissue moisture,and then treated with xylene until the tissue was transparent.The levels of oxidative stress indica-tors(MDA/GSH-Px/SOD)in mouse hippocampus were detected by ELISA;the mRNA expression levels of BDNF/TrKb/CREB were detected by RT-PCR;the protein expression levels of BDNF/TrKb/CREB were detected by Western Blot.Results Compared with the control group the gene expressions of BDNF mRNA,Trkb mRNA and CREB mRNA in the hippocampus of the experimental group were significantly increased(P＜0.05).The protein expression levels of BDNF,Trkb and CREB in the experimental group were significantly higher than those in the control group(P＜0.05).Compared with the control group,the level of MDA in the hippocampus of the mice in the experimental group was de-creased,and the levels of GSH-Px and SOD were increased(P＜0.05).The hippocampal tissue structure of the mice in the experimental group was complete,with neat and orderly arrangement of nerve cells,abundant blood vessels,and no inflammatory cell infiltration,edema,etc.;While there is a lot of inflammatory cell infiltration in the control group.Con-clusion Sevoflurane can up-regulate the levels of BDNF mRNA,Trkb mRNA and CREB mRNA in the hippocampus of mice,and the mechanism may be related to the improvement of the antioxidant capacity of the hippocampus of mice and the protection of the structure of the hippocampus of mice from damage.
关键词
七氟醚/小鼠/海马区/BDNF-TrkB-CREB信号通路/氧化应激/机制研究Key words
sevoflurane/mice/hippocampus/BDNF-TrkB-CREB signaling pathway/oxidative stress/mechanism re-search引用本文复制引用
出版年
2024
NETL
NSTL
万方数据