亚硒酸钠联合高压氧对颅脑损伤模型鼠细胞凋亡与病理形态的影响
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摘要
目的：观察亚硒酸钠结合高压氧对颅脑损伤模型鼠的脑组织病理形态学与神经细胞凋亡的影响,分析急性脑创伤的病理和病理生理变化,初步探讨其作用机制,并为稀土元素硒在颅脑损伤方面的治疗提供一定的理论依据。方法：40只SD大鼠随机分为对照组、脑损伤组和治疗组,治疗组又分为(亚硒酸钠组、高压氧组、亚硒酸钠联合高压氧处理组)。亚硒酸钠提前一月进行喂养,然后应用Feeney改良自由落体颅脑损伤装置,参照中度损伤模型的相应指标,建立大鼠闭合性颅脑损伤模型,最后以NSS再将损伤程度统一为中度颅脑损伤程度。涉及高压氧的组别伤后1h、24h、48h做高压氧治疗,各组均以72小时统一留取标本,分别测定脑组织含水量；石蜡切片HE染色光镜下观察大脑皮层以海马CA1区形态变化；TUNEL法检测各组大鼠大脑皮质凋亡细胞表达；免疫组化法检测各组大鼠大脑皮质与海马CA1区caspase-3细胞数的表达；电镜下观察各组大鼠大脑皮质与海马CA1区超微结果变化。数据采用SPSS13.0软件处理,统计处理组间比较采用。One-wayANOVA,相关性分析采用Spearman法。P<0.05为差异有显著性。结果：①脑含水量测定显示：脑损伤组和对照组大鼠比较,脑含水量明显增加(P<0.01)；治疗组与脑损伤组大鼠比较,脑含水量有不同程度的下降(P<0.05),其中联合处理组的下降幅度最多。②光镜下显示：脑损伤组和对照组比较,皮质与海马区胶质细胞显著增多；治疗组和脑损伤组比较,胶质细胞均有不同程度的减少,联合处理组减少最为明显。③细胞凋亡测定：空白组仅有极少数神经细胞呈TUNEL阳性。脑损伤组和对照组比较,皮质凋亡神经细胞数显著增多(P<0.01)；治疗组和脑损伤组比较,均有不同程度的减少(P<0.05),联合处理组减少最为明显(P<0.01)。④Caspase-3阳性细胞表达：空白组仅有极少数Caspase-3阳性细胞表达。脑损伤组和对照组比较,皮质和海马区阳性细胞数显著增多(P<0.01)；治疗组和脑损伤组比较,均有不同程度的减少(P<0.05),联合处理组减少最为明显(P<0.01)。⑤电镜超微结构变化显示：脑损伤组和对照组比较,皮质与海马区细胞结构破坏严重；治疗组和脑损伤组比较,程度减轻,联合处理组最为明显。结论：①亚硒酸钠与高压氧处理均能不同程度的抑制颅脑损伤后引起的神经细胞凋亡。②联合处理组的效果优于两种单一的处理组,说明两种处理方法有协同治疗作用。③亚硒酸钠在颅脑损伤后的病理改变中有一定的保护性作用。
bjective:To observe the effects of sodium selenite combined with hyperbaric oxygen on the rat model of traumatic brain injury in pathological brain tissue and nerve cell apoptosis, analysis pathology and pathophysiology of acute brain trauma coupled with initially investigate its mechanism, and provide a theoretical basis for the rare earth elements of selenium in the treatment of traumatic brain injury. It has to enhance the layer of treatment of acute brain injury. Methods:Fifty adult male SD rats were randomly divided into five groups(n=10 each).The five groups were treated differently as followings: shaminjured group(n=10)、trauma group(n=10)、TBI+selenium group (n=10)、TBI+HBO group(n=10)、TBI+selenium+HBO group(n=10).The experimental TBI model was established by Feeney's method except sham-injured group, followed by NSS score. The groups which blended with selenium were feeded on Sodium Salinity from one month ago. The else groups which combined with HBO therapy, meanwhile, were regularly taken HBO treatment at 1、24 and 48h after TBI.The brain samples were harvested at 72h after TBI for measuring brain water content、apoptosis cells by TUNEL and caspase-3 Protein by immuno-chemical method. The areas of cortex and hippocampus CA1 were observed,utilizing light microscope and electron microscope. All values were expressed as meansd士SD. One-way analysis of variance was used to assess group means, Spearman method to correlation analysis. P<0.05 was considered statistically significant. Statistics were using SPSS13.0. Results:①Brain water content:The water content in the trauma Group were obviously increased than that in the Shamed-operation Group(P<0.01). But the water content in the therapy Groups reduced obviously (p<0.05) than that in the trauma Group, especially in the combined treatment Group.②Displaying under the Light microscope:The glia cells of cortex and hippocampus CA1 in the trauma Group were obviously increased than that in the Shamed-operation Group(P<0.01),but the ones in the therapy Groups reduced obviously (p<0.05) than that in the trauma Group, especially in the combined treatment Group.③TUNEL Staining:there are few TUNEL-positive neurons in Shamed-operation group.After TBI, the TUNEL-positive neuronal cells increased significantly in cortex and hippocampus CA1. Therapy Group sharply decreased the number of TUNEL-positive neuronal cells induced by TBI accordingly index respectively compared with trauma Group, which is, however,more than that of Shamed-operation group(P<0.05).④Expression of caspase-3:Caspase-3 expression in cerebrum increased significantly after TBI,especially in the areas surrounding the injured cortex and hippocampus CA1. The results of the Caspase-3 positive cells in cortex and hippocampus CA1 decreased in therapy groups including TBI+selenium group、TBI+HBO group、TBI+selenium+HBO group.According to the results of trauma Group, both selenium and HBO can suppressed the expression of caspase-3 caused by TBI, especially in the combined treatment Group.⑤Displaying under the Electron microscope:The normal cell structures of cortex and hippocampus CA1 in the trauma Group were gravely damaged than that in the Shamed-operation Group(P<0.01),but the ones in the therapy Groups recovered obviously (p<0.05) than that in the trauma Group,especially in the combined treatment Group. Conclusion:①It is objectively predicted that Both Sodium Selenite and HBO can of inhibited nervous cell apoptosis caused by brain injury in the varying degrees②The very fact that combined treatment group is better than the two kinds of single one regarded two kinds of treatment as a effectively synergistic therapeutic measure.③Sodium selenite partly plays a protective effect on certain pathological changes in the brain injury.
bjective:To observe the effects of sodium selenite combined with hyperbaric oxygen on the rat model of traumatic brain injury in pathological brain tissue and nerve cell apoptosis, analysis pathology and pathophysiology of acute brain trauma coupled with initially investigate its mechanism, and provide a theoretical basis for the rare earth elements of selenium in the treatment of traumatic brain injury. It has to enhance the layer of treatment of acute brain injury. Methods:Fifty adult male SD rats were randomly divided into five groups(n=10 each).The five groups were treated differently as followings: shaminjured group(n=10)、trauma group(n=10)、TBI+selenium group (n=10)、TBI+HBO group(n=10)、TBI+selenium+HBO group(n=10).The experimental TBI model was established by Feeney's method except sham-injured group, followed by NSS score. The groups which blended with selenium were feeded on Sodium Salinity from one month ago. The else groups which combined with HBO therapy, meanwhile, were regularly taken HBO treatment at 1、24 and 48h after TBI.The brain samples were harvested at 72h after TBI for measuring brain water content、apoptosis cells by TUNEL and caspase-3 Protein by immuno-chemical method. The areas of cortex and hippocampus CA1 were observed,utilizing light microscope and electron microscope. All values were expressed as meansd士SD. One-way analysis of variance was used to assess group means, Spearman method to correlation analysis. P<0.05 was considered statistically significant. Statistics were using SPSS13.0. Results:①Brain water content:The water content in the trauma Group were obviously increased than that in the Shamed-operation Group(P<0.01). But the water content in the therapy Groups reduced obviously (p<0.05) than that in the trauma Group, especially in the combined treatment Group.②Displaying under the Light microscope:The glia cells of cortex and hippocampus CA1 in the trauma Group were obviously increased than that in the Shamed-operation Group(P<0.01),but the ones in the therapy Groups reduced obviously (p<0.05) than that in the trauma Group, especially in the combined treatment Group.③TUNEL Staining:there are few TUNEL-positive neurons in Shamed-operation group.After TBI, the TUNEL-positive neuronal cells increased significantly in cortex and hippocampus CA1. Therapy Group sharply decreased the number of TUNEL-positive neuronal cells induced by TBI accordingly index respectively compared with trauma Group, which is, however,more than that of Shamed-operation group(P<0.05).④Expression of caspase-3:Caspase-3 expression in cerebrum increased significantly after TBI,especially in the areas surrounding the injured cortex and hippocampus CA1. The results of the Caspase-3 positive cells in cortex and hippocampus CA1 decreased in therapy groups including TBI+selenium group、TBI+HBO group、TBI+selenium+HBO group.According to the results of trauma Group, both selenium and HBO can suppressed the expression of caspase-3 caused by TBI, especially in the combined treatment Group.⑤Displaying under the Electron microscope:The normal cell structures of cortex and hippocampus CA1 in the trauma Group were gravely damaged than that in the Shamed-operation Group(P<0.01),but the ones in the therapy Groups recovered obviously (p<0.05) than that in the trauma Group,especially in the combined treatment Group. Conclusion:①It is objectively predicted that Both Sodium Selenite and HBO can of inhibited nervous cell apoptosis caused by brain injury in the varying degrees②The very fact that combined treatment group is better than the two kinds of single one regarded two kinds of treatment as a effectively synergistic therapeutic measure.③Sodium selenite partly plays a protective effect on certain pathological changes in the brain injury.
引文
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