国际麻醉学与复苏杂志   2014, Issue (5): 10-10
    
异氟烷对血管性痴呆老年大鼠海马中胆碱能系统的影响
huzhichao, 许鹏程1()
1.河南省商丘市第一人民医院麻醉科
Effects of isoflurane on the cholinergic system in hippocampus in aged rats with vascular dementia
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摘要:

目的 研究异氟醚对血管性痴呆(vascular dementia, VaD)老年大鼠海马中胆碱能系统的影响。 方法 采用四血管法(4-vessel occlusion, 4-VO)制作VaD老年(15月龄)大鼠模型,选用60只SD老年大鼠,雌雄各半,制作成VaD老年大鼠。根据异氟烷吸入浓度(0、1.5%、2.0%)和吸入后开始测试胆碱能递质的天数(1 d和7 d)将上述大鼠分为6组,即:1 d未吸入异氟烷组(Ⅰ1,0),1 d吸入1.5%异氟烷组(Ⅰ1,1.5%),1 d吸入2.0%异氟烷组(Ⅰ1,2.0%),7 d未吸入异氟烷组(Ⅰ7,0),7 d吸入1.5%异氟烷组(Ⅰ7,1.5%),7 d吸入2.0%异氟烷组(Ⅰ7,2.0%)。吸入异氟烷后在第1天和第7天分别进行Morris水迷宫测试。水迷宫测试完成后,立即处死大鼠,在冰面取出大鼠的海马,放入-80 ℃的冰箱中保存。然后检测海马中的乙酰胆碱含量、乙酰胆碱转移酶及乙酰胆碱酯酶的活力。 结果 吸入1.5%和2.0%异氟烷1 d后,海马中乙酰胆碱含量[(82±10) μg/g和(65±13) μg/g]和乙酰胆碱转移酶活力[(98±11) IU/g和(81±13) IU/g]均明显低于未吸入异氟烷组[乙酰胆碱含量(99±13) μg/g和乙酰胆碱转移酶(115±11) IU/g],吸入2.0%异氟烷模型大鼠下降更为明显(P<0.05);随着吸入异氟烷浓度不同而乙酰胆碱酯酶[(0.75±0.33)、(0.85±0.35) U/mgprot和(0.55±0.22) U/mgprot]的变化差异无统计学意义(P>0.05)。模型大鼠吸入异氟烷后第7天,各组大鼠海马中乙酰胆碱含量[(99±13)、(97±13) μg/g和(98±12) μg/g]和乙酰胆碱转移酶[(118±12)、(128±15) IU/g和(116±15) IU/g]及乙酰胆碱酯酶活力[(0.71±0.29)、(0.70±0.11) U/mgprot和(0.66±0.38) U/mgprot]没有明显变化(P>0.05)。 结论 异氟烷能使VaD老年大鼠海马中乙酰胆碱含量及乙酰胆碱转移酶活力短暂下降 ,而对乙酰胆碱酯酶无明显影响。

关键词: 血管性痴呆; 老年; 异氟烷; 学习记忆; 乙酰胆碱; 乙酰胆碱转移酶; 乙酰胆
Abstract:

Objective To investigate the effects of isoflurane at different concentrations on the cholinergic system on the hippocampus of aged rats with vascular dementia(VaD). Methods The model of the old VaD rats was established with the method of 4-vessel occlusion(4-VO) in 60 aged SD rats (30 males and 30 females). The animals were divided into 6 groups according to the days(days 1 and 7)after isoflurane inhalation and the concertrations(0, 1.5% and 2.0%):1 d 0 isoflurane group(Ⅰ1,0), 1 d 1.5% isoflurane group(Ⅰ1,1.5%),1 d 2.0% isoflurane group(Ⅰ1,2.0%),7 d 0 isoflurane group(Ⅰ7,0),7 d 1.5% isoflurane group(Ⅰ7,1.5%),7 d 2.0% isoflurane group(Ⅰ7,2.0%). On day 1 and day 7 after isoflurane inhalation,morris water maze testing was performed. After Morris water maze testing was completed,the aged rats with VaD were killed at once. Then the hippocampus of the aged rats with VaD were dislodged on ice and reserved in icebox at subzero 80 temperature with iquid nitrogen frozening in order to detect the content of acetylcholine,choline acetyltransferase and acetylcholinesterase. The content of acetylcholine in hippocampus is detected by Elisa method,and the content of choline acetyltransferase is detected by kit of choline acetyltransferase and the content of acetylcholinesterase is detected by kit of acetylcholinesterase. Results On the 1st day after isoflurane inhalation,the content of acetylcholine [Ⅰ1,1.5%(82±10) μg/g and Ⅰ1,2.0%(65±13) μg/g] and choline acetyltransferase [Ⅰ1,1.5%(98±11) IU/g and Ⅰ1,2.0%(81±13) IU/g] in hippocampus were also markedly lower than those of 0 isoflurane group[the content of acetylcholine (99±13) μg/g and the content of choline acetyltransferase (115±11) IU/g](P<0.05),of which the 2.0% isoflurane group had the lowest(P<0.05). There were not significant different of acetylcholinesterase with different concertrations of isoflurane inhalation(P>0.05). On the 7th day after isoflurane inhalations, no significant differences were observed the content of acetylcholine[(99±13),(97±13) μg/g and (98±12) μg/g], choline acetyltransferase [(118±12),(128±15) IU/g and (116±15) IU/g] and acetylcholinesterase [(0.71±0.29),(0.70±0.11) U/mgprot and(0.66±0.38) U/mgprot] in hippocampus among three groups(P>0.05). Conclusions After VaD rats suck in isoflurane, the content of acetylcholine and choline acetyltransferase in the hippocampus were decreased but only temporary, but the contents of acetylcholinesterase was not influenced.

Key words: Vascular dementia; Old age; Isoflurane; Learning and memory; Acetylcholine; Choline acetyltransferase; Acetylcholinesterase; Hippocampus