海洛因成瘾大鼠脑组织超微结构和GFAP表达的变化
摘要
目的：模仿人类吸毒成瘾的临床实例，建立海洛因成瘾大白鼠模型，研究海洛因成瘾大白鼠脑神经组织超微结构及GFAP的表达变化，了解海洛因成瘾对大白鼠脑的损害作用。方法：采用递增法人为地给大白鼠皮下注射海洛因造成动物成瘾，动物设对照组和模型组，取2组大白鼠全脑作正中矢状切分为两半，半边脑做胶质原纤维酸性蛋白(GFAP)免疫反应，另半边脑取前额皮质、海马、下丘脑等部位脑组织做电镜观察。结果：海洛因成瘾大白鼠脑内多部位神经元胞体、轴突、树突都出现变性、凋亡、胀亡等超微病理结构改变；星形胶质细胞增生肥大，亦可见退变星形胶质细胞，少突胶质细胞、小胶质细胞也增生；神经毯出现肿胀、变性等超微病理结构改变；广泛性出现血脑屏障(BBB)通透性增加的超微结构变化，BBB周围星形胶质细胞水肿； GFAP表达水平明显增强；上述病理改变未显示出部位上的特异性。对照组电镜超微结构影像和GFAP表达水平正常。结论：海洛因成瘾大白鼠脑组织出现广泛性损害，神经元胞体与突起、神经胶质细胞、神经毯、血脑屏障(BBB) 均出现超微病理结构改变，其中神经元以变性为主，神经元凋亡是海洛因成瘾致脑神经元死亡的主要形式，星形胶质细胞明显增生肥大，GFAP表达水平明显增强，本实验结果未显示出海洛因成瘾对脑组织损害有部位上的特异性。
Objective: To imitate the processes of human heroin addiction, we set up the model of heroin addiction. The study on the changes of ultrastructune and expression of GFAP in the brain tissues of heroin-addicted rats were done to make clear the lesions which have been made in the brain tissues by heroin. Method: Appling the method of gradually increase of dosage, rats were given subcutaneous injection of heroin and the addiction of model was established. Rats were divided into control group and model group. The brains were taken out from two groups rats and were divided into two halfs along the median sagittal plane. The expression of GFAP were tested by immnohistochemical method in the half brain. The ultrastructural changes of brain tissues in the prefroneal cortex hippocampus and hypothalamus which were taken out from the another half brain were observed under eletronic microscope. Results: Degeneration, apoptosis, oncosis were found in the neuronal somas, axons, dendrites of the heroin addicted rats brains. Astrocytes were hyperplasia and hypertrophy. Meanwhile, the degenerative astrocytes were also found. Oligodenodrocytes and microglial cell were hyperplasia too. The swellon and degenernatial neuropil were found. A series of ultrastructural changes of permeability-increase of blood-brain barrier (BBB) of heroin-addicted rats existed. The glial perivascular feet and astrocytes surrounding the basement membrance were swellon. Expression levels of GFAP increased sighificantly. There was not position differentiae among these pathological changes in this experience. Ultrastructural image and expression
levels of GFAP in the brain of control group were normal. Conclusion: Widespread pathology ultrastruceural changes have been found in the heroin-addicted rats brain tissue, including neuronal somas and processes, glial cells ,neuropil and BBB, of which degeneration is a main form of pathology and apoptosis is the main form of neuron death .Accordingly,astrocytes were hyperplasia and expression level of GFAP increased significantly. This experim ent didn’t demonstrated that there was position differentiae in the lesions of brain tissue which were made by heroin addiltion.
Objective: To imitate the processes of human heroin addiction, we set up the model of heroin addiction. The study on the changes of ultrastructune and expression of GFAP in the brain tissues of heroin-addicted rats were done to make clear the lesions which have been made in the brain tissues by heroin. Method: Appling the method of gradually increase of dosage, rats were given subcutaneous injection of heroin and the addiction of model was established. Rats were divided into control group and model group. The brains were taken out from two groups rats and were divided into two halfs along the median sagittal plane. The expression of GFAP were tested by immnohistochemical method in the half brain. The ultrastructural changes of brain tissues in the prefroneal cortex hippocampus and hypothalamus which were taken out from the another half brain were observed under eletronic microscope. Results: Degeneration, apoptosis, oncosis were found in the neuronal somas, axons, dendrites of the heroin addicted rats brains. Astrocytes were hyperplasia and hypertrophy. Meanwhile, the degenerative astrocytes were also found. Oligodenodrocytes and microglial cell were hyperplasia too. The swellon and degenernatial neuropil were found. A series of ultrastructural changes of permeability-increase of blood-brain barrier (BBB) of heroin-addicted rats existed. The glial perivascular feet and astrocytes surrounding the basement membrance were swellon. Expression levels of GFAP increased sighificantly. There was not position differentiae among these pathological changes in this experience. Ultrastructural image and expression
levels of GFAP in the brain of control group were normal. Conclusion: Widespread pathology ultrastruceural changes have been found in the heroin-addicted rats brain tissue, including neuronal somas and processes, glial cells ,neuropil and BBB, of which degeneration is a main form of pathology and apoptosis is the main form of neuron death .Accordingly,astrocytes were hyperplasia and expression level of GFAP increased significantly. This experim ent didn’t demonstrated that there was position differentiae in the lesions of brain tissue which were made by heroin addiltion.
引文
1. 姜佐宁主编. 药物依赖—临床特征与现代治疗(第二版) .北京:人民卫生出版社,2003,3:8-23.
2. 张锐敏主编.海洛因等阿片类物质依赖的临床与治疗.太原:山西科学技术出版社,1996,6:7-108.
3. 秦伯益.阿片受体与阿片类依赖的防治. 中国新药杂志,1995,4(6):3-6.
4.Koob GF, Sanna PP, Bloom FE. Neuroscience of addition. Nenrm.1998,21:467-476.
5. Buttner A. Mall G. Penning R. et al. The Neuro pathology of heroin abuse. Forensie sci Int 2000,113(1-3):435-442.
6.张开镐.药物依赖性的动物实验方法(一).中国药物依赖性杂志，1999,8(1):23-26.
7.包新民,舒斯云.大鼠脑立体定位图谱.北京:人民卫生出版社,1991.
8. Maldonado R, Negus S, Koob GF. Precipitation of morphine withdrawal syndrome in rats by administration of mu-delta-and Kappa-selective opioid antagonists [J]. NeuroPharmacology, 1992, 31(12):1231~1241.
9. 徐贵丽, 刘泽源, 苏国丽,等. 大鼠海洛因依赖催促戒断模型的建立[J]. 中国药物滥用防治杂志, 2000, (3)2:9~10.
10. 武忠弼主编.超微病理诊断学.上海:上海科学技术出版社,2003: 396-443.
11.季凤清.缺氧复氧诱导大鼠大脑皮层神经细胞凋亡的形态学研究.解剖科学进展.2001,7（4）：213-215.
12.Li Y, Sharor VG, Jiang N. ultrastructural and ligt microscopic evidence of apoptosis after middle cerebral artery occlusion in the rat. Am J pathol, 1995,146:1045-1051.
13.Majno G,Joris I.Apoptosis,oncosis,and necrosis.A overview of cell death.Am J Pathol,1995,146:3-15.
14.吴人亮，李娜萍．细胞凋亡、胀亡和坏死－关于细胞死亡的新认识．中华病理学杂志，2002,31(5):255-256.
15.李利华.海洛因成瘾者神经内分泌及免疫系统超微病理变化研究.法医学杂志,2001,17(3):145-147.
16.徐建春.海洛因依赖及戒断后雄性大鼠性腺轴组织病理学研究. 中国药物依赖通报.1997,6(3):140-144.
17.阮奕文,姚志彬.老年大鼠蓝斑胶质细胞超微结构的变化.广东解剖学通报,1995,17(1):31-34.
18.Kajhara H, Tsutsumi E, Kinoshita A, et al. Activated astrocytes with glycogen accumulation in ischemic penumbra during the early stade of brain infarction immunohiseochtmical and electon microscopic studies. Braiu Res, 2001,909:92-101.
19.Wang LC.Astroglial differentiation is required for support of rewrite outgrowth. J Nenrosci. 1994,14(5):3195.
20.刘静,贾克勇,李玲等.不同声压级次声作用小鼠后脑胶质纤维酸性蛋白的含量和意义.中国临床康复.2001,5(7):13-15.
21.马建荣,姜冰,马志明.正常大白鼠经伽玛刀照射后脑组织胶质纤维酸性蛋白的变化.湖南医科大学学报.2001,26(4):6-8
22.Nawashiro H, Messing A, Azzam N. Mice lacking GFAP are hypersensitivity to traumatic cerebrospinal injury. Nenro Report, 1998,19(8):1691
23.Hinkle DA. Baldwin SA, Scheff SW, et al. GFAP and S-100 beta expression in the cortex and hippocampus in response to wild cortical
contusion. J Neurotrauma, 1997,14(10):729.
24.Tanaka H, Katoh A, Oguro K, et al. Disturborce of hippocampus lang-term potentiation after transient ischemia in GFAP deficient Mice. J Neurosi Res, 2002,67:11-20.
25.Temburni MK, Jacob MH. New function for glia in the brain [J]. Proc Natl Acad Sci USA, 2001,98(7):3631-3632.
26.张光鹏,羊惠君,袁琼兰等.大鼠脑缺血再灌注对神经元损伤与星形胶质细胞的反应.神经解剖学杂志,2000,16(3):
27.段德新,刘之荣.缺血性脑损害与胶质细胞反应.国外医学(老年医学分册),2000,21(5):42-44
28.李霞斯,卞晓红,刘之荣.慢性脑灌注不足脑内小胶质细胞的反应.西北国防医学杂志.2000,21(2):23-25.
29.贾克勇,陈景藻,刘先珍等.次声作用后鼠脑超微结构与血脑屏障的改变.中华生物医学康复杂志,1999,21(3):30-33.
30.闵珍,陆纯英,顾祖维等.低浓度铅对大鼠血脑屏障超微结构的损害.中华劳动卫生职业病杂志,1998,16(6):8-11.
31.张开镐.药物成瘾的病理生理学基础.中国药物依赖性杂志,2002,11(2):81-85.
32.Nestler EJ, Aghajamian GK. Molecular and cellular basis of addiction. Science, 1997:278-303.
33.Pizza PV, Moal ML. Pathophysiological basis of vulerability to drug abuse role of an ineeration between stress, glucocorticoide and dopaminergic neurons. Am Rev Pharmacol Toxicol, 1996,36:359.
2. 张锐敏主编.海洛因等阿片类物质依赖的临床与治疗.太原:山西科学技术出版社,1996,6:7-108.
3. 秦伯益.阿片受体与阿片类依赖的防治. 中国新药杂志,1995,4(6):3-6.
4.Koob GF, Sanna PP, Bloom FE. Neuroscience of addition. Nenrm.1998,21:467-476.
5. Buttner A. Mall G. Penning R. et al. The Neuro pathology of heroin abuse. Forensie sci Int 2000,113(1-3):435-442.
6.张开镐.药物依赖性的动物实验方法(一).中国药物依赖性杂志，1999,8(1):23-26.
7.包新民,舒斯云.大鼠脑立体定位图谱.北京:人民卫生出版社,1991.
8. Maldonado R, Negus S, Koob GF. Precipitation of morphine withdrawal syndrome in rats by administration of mu-delta-and Kappa-selective opioid antagonists [J]. NeuroPharmacology, 1992, 31(12):1231~1241.
9. 徐贵丽, 刘泽源, 苏国丽,等. 大鼠海洛因依赖催促戒断模型的建立[J]. 中国药物滥用防治杂志, 2000, (3)2:9~10.
10. 武忠弼主编.超微病理诊断学.上海:上海科学技术出版社,2003: 396-443.
11.季凤清.缺氧复氧诱导大鼠大脑皮层神经细胞凋亡的形态学研究.解剖科学进展.2001,7（4）：213-215.
12.Li Y, Sharor VG, Jiang N. ultrastructural and ligt microscopic evidence of apoptosis after middle cerebral artery occlusion in the rat. Am J pathol, 1995,146:1045-1051.
13.Majno G,Joris I.Apoptosis,oncosis,and necrosis.A overview of cell death.Am J Pathol,1995,146:3-15.
14.吴人亮，李娜萍．细胞凋亡、胀亡和坏死－关于细胞死亡的新认识．中华病理学杂志，2002,31(5):255-256.
15.李利华.海洛因成瘾者神经内分泌及免疫系统超微病理变化研究.法医学杂志,2001,17(3):145-147.
16.徐建春.海洛因依赖及戒断后雄性大鼠性腺轴组织病理学研究. 中国药物依赖通报.1997,6(3):140-144.
17.阮奕文,姚志彬.老年大鼠蓝斑胶质细胞超微结构的变化.广东解剖学通报,1995,17(1):31-34.
18.Kajhara H, Tsutsumi E, Kinoshita A, et al. Activated astrocytes with glycogen accumulation in ischemic penumbra during the early stade of brain infarction immunohiseochtmical and electon microscopic studies. Braiu Res, 2001,909:92-101.
19.Wang LC.Astroglial differentiation is required for support of rewrite outgrowth. J Nenrosci. 1994,14(5):3195.
20.刘静,贾克勇,李玲等.不同声压级次声作用小鼠后脑胶质纤维酸性蛋白的含量和意义.中国临床康复.2001,5(7):13-15.
21.马建荣,姜冰,马志明.正常大白鼠经伽玛刀照射后脑组织胶质纤维酸性蛋白的变化.湖南医科大学学报.2001,26(4):6-8
22.Nawashiro H, Messing A, Azzam N. Mice lacking GFAP are hypersensitivity to traumatic cerebrospinal injury. Nenro Report, 1998,19(8):1691
23.Hinkle DA. Baldwin SA, Scheff SW, et al. GFAP and S-100 beta expression in the cortex and hippocampus in response to wild cortical
contusion. J Neurotrauma, 1997,14(10):729.
24.Tanaka H, Katoh A, Oguro K, et al. Disturborce of hippocampus lang-term potentiation after transient ischemia in GFAP deficient Mice. J Neurosi Res, 2002,67:11-20.
25.Temburni MK, Jacob MH. New function for glia in the brain [J]. Proc Natl Acad Sci USA, 2001,98(7):3631-3632.
26.张光鹏,羊惠君,袁琼兰等.大鼠脑缺血再灌注对神经元损伤与星形胶质细胞的反应.神经解剖学杂志,2000,16(3):
27.段德新,刘之荣.缺血性脑损害与胶质细胞反应.国外医学(老年医学分册),2000,21(5):42-44
28.李霞斯,卞晓红,刘之荣.慢性脑灌注不足脑内小胶质细胞的反应.西北国防医学杂志.2000,21(2):23-25.
29.贾克勇,陈景藻,刘先珍等.次声作用后鼠脑超微结构与血脑屏障的改变.中华生物医学康复杂志,1999,21(3):30-33.
30.闵珍,陆纯英,顾祖维等.低浓度铅对大鼠血脑屏障超微结构的损害.中华劳动卫生职业病杂志,1998,16(6):8-11.
31.张开镐.药物成瘾的病理生理学基础.中国药物依赖性杂志,2002,11(2):81-85.
32.Nestler EJ, Aghajamian GK. Molecular and cellular basis of addiction. Science, 1997:278-303.
33.Pizza PV, Moal ML. Pathophysiological basis of vulerability to drug abuse role of an ineeration between stress, glucocorticoide and dopaminergic neurons. Am Rev Pharmacol Toxicol, 1996,36:359.