乙醇灌胃对大鼠海马区胶质纤维酸性蛋白、S-100B蛋白表达的影响

Effect of gastric administration of alcohol on the expressions of glial fibrillary acidic protein and S-100B protein in hippocampus of rats

目的:观察乙醇灌胃致大鼠海马区神经胶质细胞内胶质纤维酸性蛋白和S-100B蛋白表达的改变。方法:实验于2004-09/2005-09在延边大学医学院附属医院神经内科学教研室完成。选用3月龄Wistar雄性大鼠60只,完全随机分为正常组、灌胃1,2,4,6,8周组6组,每组10只,各灌胃组按8g/(kg·d)乙醇灌胃,依周数喂养结束后取大鼠脑组织,常规制片,行胶质纤维酸性蛋白和S-100B蛋白的免疫组化染色,光镜下进行形态学观察;采用HPIAS-1000计算机彩色病理图文分析系统对胶质纤维酸性蛋白和S-100B阳性细胞计数行定量分析,进行各灌胃组与正常组的比较。结果:60只Wistar大鼠中灌胃8周组于喂养第7周死亡1只(死亡原因为乙醇误灌入肺内,造成急性肺水肿而死亡),其余各组动物全部进入结果分析。①形态学改变:正常神经胶质细胞胞体轮廓清晰,星状突起纤细,胶质纤维酸性蛋白分布于胞浆及星状突起中。S-100B阳性染色也见于胶质细胞中,均匀分布于胞浆中。随灌胃周数增加,胶质纤维酸性蛋白阳性细胞胞体变大、形态各异,突起增多明显且粗大不规则,S-100B阳性细胞体积有所增大,胞浆丰富显色较深。②灌胃1,2,4,6,8组大鼠胶质纤维酸性蛋白和S-100B免疫反应产物阳性细胞计数值均较正常组显著增高,差异有显著性意义犤胶质纤维酸性蛋白阳性细胞计数:(76.49±6.43),(84.20±3.17),(92.60±4.81),(138.20±5.52),(142.12±6.41),(29.32±2.42)个,F=883.62,P<0.05犦;犤S-100B阳性细胞计数:(196.96±11.47),(182.22±17.28),(215.88±15.50),(239.70±14.95),(254.92±23.42),(115.53±11.62)个,F=99.88,P<0.05犦。结论:乙醇使大鼠海马区胶质纤维酸性蛋白和S-100B的表达增加,增加的程度与乙醇灌胃时间有关。随着灌胃时间的延长,大鼠海马区胶质纤维酸性蛋白和S-100B的表达增多。

AIM： To investigate the effects of intragastric administration of alcohol on protein expression changes inglial fibrillary acidic protein （GFAP） and S-100B in neuroglia cells of rat hippocampus. METHODS： The experiment was performed in the Department of Neurology, Affiliated Hospital of Medical College of YaMbian University between September 2004 and September 2005. Sixty male Wistar rats of three-month-old of rats were selected and randomly divided into six groups with 10 rats in each group： normal group, 1-week, 2-week, 4-week, 6-week, 8-week perfusion group. Rats in all perfusion groups were given intragastric administration of alcohol group at 8 g/kg per day, and the brain tissues of rats were obtained according to the feeding time, which were then routinely made into slices to immunohistochemically stain GFAP and S-100B. The morphological changes were studied under optical microscope. The number of positive cells of GFAP and S-100B was analyzed by HPIAS-1000 analysis system, and the those in all perfusion groups were compared with those in the normal group. RESULTS： Of 60 Wistar rats, one rat in the 8-week perfusion group died in the 7^th week （died for acute pulmonary edema caused by mis-perfusion of alcohol into the lung）, and all the rests were involved in the analysis of results.①The changes in morphology： The figures of normal neuroglia cells were clear, and the stellate prominences were slim. GFAP was distributed among cytoplasm and stellate prominences. The positive staining of S-100B was also found in neuroglial cells with uniform distribution in cytoplasm. With the prolongation of time, the positive cells of GFAP became big, which were different from each other in morphology. The positive cells volume of S-100B was bigger, and the staining of cytoplasm was darker than ever.②The number of immunoreaetions positive cells of GFAP and S-100B in all perfusion groups were significantly increased than that in the normal group [number of positive GFAP cells： （76.49±6.43）, （84.20±3.17）, （92.60±4.81）, （138.20±5.52）, （ 142.12±6.41 ）, （29.32±2.42）, F=883.62,P 〈 0.05]; [number of positive cells of S-100B： （196.96±11.47）, （ 182.22 ±17.28）, （215.88±15.50）, （239.70±14.95）, （254.92 ±23.42）, （115.53 ± 11.62）, F=99.88 ,P 〈 0.05]. CONCLUSION： Intragastric administration of alcohol can increase the protein expressions of GFAP and S-100B in rat hippocampus, and the increase is related with the time of perfusion. With the time of perfusion increases, the expressions of GFAP and S-100B inerease in rat hippocampus.