大鼠阻塞性睡眠呼吸暂停低通气综合征动物模型的建立

发布时间：2018-01-02 19:26

本文关键词：大鼠阻塞性睡眠呼吸暂停低通气综合征动物模型的建立　出处：《山东大学》2009年硕士论文　论文类型：学位论文

【摘要】： 目的:阻塞性睡眠呼吸暂停低通气综合征(obstructive sleep apnea hypopneasyndrome,OSAHS)是一种临床上的常见疾病,是睡眠时上呼吸道反复塌陷阻塞所引起的呼吸暂停和低通气,并出现睡眠时打鼾,白天嗜睡,睡眠结构改变和睡眠时血氧饱和度下降,累及全身各系统、多脏器,对人体健康造成严重危害,已引起国内外学者的广泛重视。本实验的目的是建立一个稳定、可靠的OSAHS动物模型,以进一步研究OSAHS的发病机制、病理生理改变和并发症。方法: 1.麻醉:将戊巴比妥钠用无菌生理盐水配成0.8%的戊巴比妥钠溶液,即配即用。手术操作和睡眠监测时分别采用不同剂量作腹腔内注射。 2.咽部注入透明质酸钠:大鼠麻醉后,在双侧舌腭弓、咽腭弓及舌根处注射透明质酸钠至大鼠出现呼吸暂停。 3.信号的采集:以经过改造的多导睡眠仪监测动物的脑电(electroencephalogram,EEG)、肌电(electromyogram,EMG)和口鼻气流(oronasalairflow,AIRFLOW)。 4.信号的分析:人工分析所监测到的脑电、肌电和口鼻气流。 4.1清醒与睡眠的区分:以多导睡眠仪监测脑电和肌电,采取以下方法判断动物是否入睡:清醒状态:脑电波为高频率,低幅度,有较高幅度的肌电活动;睡眠状态:脑电波为低频率,高幅度,同时伴有肌电活动幅度下降。 4.2呼吸暂停:以呼吸波消失≥2.5s作为一次呼吸暂停,计数每只动物于清醒期和睡眠期的呼吸暂停次数并计算每只动物的呼吸暂停指数(apnea index,AI) 4.3最长呼吸暂停时间(longest apnea time,Ltime):分析每只动物在清醒期和睡眠期的最长呼吸暂停时间。 5.统计学处理:采用SPASS10.0统计软件进行数据分析,注射前后动物AI及Ltime的比较使用配对t检验,清醒期与睡眠期AI及Ltime比较亦采用配对t检验。结果: 1.一般情况:注射后动物进食正常,注射后4周动物均成活,未见咽腔感染征象。 2.动物处于浅麻醉状态时,经过一定时间的潜伏期后,脑电和肌电活动与人类非快速动眼睡眠期(non-rapid eye movement,NREM)相似,即动物于麻醉状态时,脑电图呈现高幅,低频的特点,同时伴有肌电活动幅度的下降。本次实验没有监测到类似快速动眼睡眠期(rapid eye movement,REM)的脑电和肌电特点,即脑电图呈低幅,高频,肌电活动极度下降。 3.注射前和注射后4周,动物在监测过程中均出现伴有胸腹反常运动的呼吸暂停,即阻塞性睡眠呼吸暂停,但是注射前后AI及Ltime之间差异有统计学意义(P＜0.05)。 4.动物于清醒期和睡眠期均出现阻塞性睡眠呼吸暂停,但是清醒期与睡眠期AI及Ltime之间差异有统计学意义(P＜0.05)。结论:咽部注射透明质酸钠法所建立的大鼠OSAHS模型稳定、可靠,很好的模拟了人类OSAHS发病机制中的上呼吸道阻塞因素,与人类OSAHS相似,可应用于OSAHS的发病机制、病理生理改变和并发症等方面的研究。
[Abstract]:Objective: obstructive sleep apnea hypopnea syndrome (obstructive sleep apnea hypopneasyndrome, OSAHS) is a common clinical disease, sleep when the upper respiratory tract repeated collapse obstructive apnea and hypopnea, and snoring, daytime sleepiness, sleep structure changes and sleep desaturation, involving the whole body system, multiple organs, causing serious harm to human health, has aroused the attention of scholars at home and abroad. The purpose of this study is to establish a stable and reliable animal model of OSAHS, in order to further study the pathogenesis of OSAHS, and the change of physiological and pathological complications.
Method:
1. anesthesia: sodium pentobarbital was added to 0.8% pentobarbital sodium solution with sterile saline, which is ready to use. During operation and sleep monitoring, different doses of intraperitoneal injection were applied respectively.
The 2. pharyngeal injection of sodium hyaluronate in rats after anesthesia, the bilateral lingual palatal arch, pharyngopalatine arch and tongue at the injection of sodium hyaluronate to rats appear apnea.
3., signal acquisition: We monitored the electroencephalogram (EEG), electromyogram (EMG) and Oro nose flow (oronasalairflow, AIRFLOW) of the animals with a modified polyphone.
4. signal analysis: artificial analysis of the monitored electroencephalogram, electromyography, and mouth and mouth airflow.
4.1 distinguish between awake and sleep: polysomnography EEG and EMG, take the following methods to judge whether the animal to sleep: awake brain waves as high frequency, low amplitude, a higher rate of EMG activity; sleep EEG: low frequency, high amplitude, accompanied by EMG activity decline.
4.2: the respiratory apnea disappeared over 2.5s wave as an apnea, counting each animal in waking and sleeping apnea times and calculation of each animal apnea index (apnea index AI)
4.3 the longest apnea time (longest apnea time, Ltime): analysis of the longest apnea time in each animal during waking and sleep periods.
5. statistical processing: data were analyzed by SPASS10.0 statistical software. Paired t test was used for comparison of AI and Ltime before and after injection, and paired t test was used for comparison between wakefulness period and sleep period AI and Ltime.
Result:
1. general situation: after the injection, the animals were eating normally, and all the animals survived for 4 weeks after the injection, and there was no sign of the pharynx infection.
2. animal in a state of light anesthesia, after a certain time after the incubation period, the EEG and EMG activity and human non REM sleep (non-rapid eye, movement, NREM) is similar to that in animal anesthesia, EEG showed high amplitude, low frequency characteristics, accompanied with the decrease amplitude of EMG activity. This experiment did not similar to the monitoring of REM sleep (rapid eye, movement, REM) of the EEG and EMG features, namely EEG showed low amplitude, high frequency, EMG activity is extremely decreased.
3. before and 4 weeks after injection, there was apnea in the process of monitoring, which was accompanied by abnormal movement of chest and abdomen, namely obstructive sleep apnea. However, there was a significant difference between AI and Ltime before and after injection (P < 0.05).
4. animals had obstructive sleep apnea during both waking and sleep periods, but there was a significant difference between AI and Ltime (P < 0.05).
Conclusion: the OSAHS rat model of pharyngeal injection of sodium hyaluronate in the stable, reliable, good simulation of the pathogenesis of human upper respiratory tract obstruction on OSAHS factors, similar to human OSAHS pathogenesis can be applied to OSAHS, pathologic changes and complications of the study.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R766.9;R-332

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