云南白药对大鼠创伤性颅脑损伤治疗作用的研究
摘要
本研究在建立稳定可靠的液压打击创伤性颅脑损伤(TBI)模型的基础上,通过云南白药对中重型颅脑损伤后大鼠神经功能损伤评分(NSS),脑水肿程度,神经细胞凋亡指数(AI)及相关基因Caspase-3表达的影响来探讨云南白药对TBI大鼠模型的治疗作用。
第一部分大鼠中重度创伤性颅脑损伤模型的建立和云南白药对大鼠创伤性颅脑损伤后神经功能及脑含水量的影响
目的在建立稳定可靠的液压打击TBI模型基础上探讨云南白药对中重度颅脑损伤后大鼠神经功能损伤评分及脑水肿程度的影响。
方法24只SD大鼠随机分为空白对照组、模型对照组、云南白药治疗组,各8只。云南白药治疗组和模型对照组采用液压打击法,造成中重度TBI模型。云南白药治疗组给予云南白药灌胃(100mg/kg体重,造模前一天与造模后5天内每天1次),空白对照组和模型对照组只给予等量温开水灌胃。术后第0天,第3天,第5天进行神经损害程度评分(NSS),第5天处死大鼠,取脑组织标本进行病理学检查,并计算干湿重比。
结果1)2.0-2.2atm致伤压力下造成的大鼠中重度TBI模型稳定可靠,死亡率可控。2)云南白药治疗组NSS评分较模型对照组在第0天无明显差别(P>0.05),在第3天,第5天显著降低,统计结果有显著性差异(P<0.05)。3)第5天云南白药治疗组脑组织病理改变及细胞形态学改变较模型对照组轻,脑含水量明显低于模型对照组,统计结果显示有显著性差异(P<0.05)。结论1)选择2.0-2.2atm致伤压力下液压打击TBI模型稳定可靠,可满足本实验研究需要。2)云南白药可以减低TBI后大鼠NSS评分,减轻脑水肿程度,对TBI具有治疗作用。
第二部分云南白药对中重度创伤性颅脑损伤后神经细胞凋亡及相关基因Caspase-3表达的影响
目的探讨云南白药对中重度颅脑损伤后大鼠神经细胞凋亡及相关基因Caspase-3表达的影响。
方法42只SD大鼠随机分组,空白对照组6只,模型对照组与云南白药治疗组各分为24h、72h、120h组,每组6只。云南白药治疗组和模型对照组采用液压打击法,造成中重度TBI模型。云南白药治疗组给予云南白药灌胃(100mg/kg体重,造模前一天与造模后每天1次),空白对照组和模型对照组只给予等量温开水灌胃。术后24h、72h、120h处死大鼠,灌注取脑组织标本免疫组化法检测皮质区和海马区细胞凋亡指数及Caspase-3的表达。
结果1)云南白药治疗组细胞凋亡指数在24h、72h、120h均低于模型对照组,且具有显著性差异(P<0.05),并明显高于空白对照组(P<0.05)。2)云南白药治疗组Caspase-3阳性细胞数在24h、72h、120h均低于模型对照组,且具有显著性差异(P<0.05),也明显高于空白对照组(P<0.05)。
结论中重度TBI后存在神经细胞凋亡和凋亡相关蛋白Caspase-3表达增加。云南白药可显著抑制TBI后大鼠神经细胞凋亡及Caspase-3基因表达,可起到对TBI的治疗作用。
On the basis of stable and reliable fluid percussing traumatic brain injury(TBI) models, we studied the therapeutic effects of Yunnan Baiyao on the model rats by testing Neurological Severity Score(NSS),cerebral edema, apoptosis index and related Caspase-3 gene expression.
Part One:The foundation of moderate or severe TBI model and the effect of Yunnan Baiyao on NSS and cerebral edema in TBI model rats.
Objective:On the basis of stable and reliable fluid percussing traumatic brain injury(TBI) models, we studied the brain protective effect of Yunnan Baiyao on the model rats by testing Neurological Severity score(NSS) and cerebral edema.
Methods:24 SD rats were randomly assigned into three groups:control group, model control group and Yunnan Baiyao(YB) treatment group. Each group contained 8 rats. Model control group and Yunnan Baiyao(YB) treatment group were established moderate or severe TBI model according to fluid percussing method. The YB treatment group was treated with Yunnan Baiyao (100mg/kg,q.d) and warm water, the control group and model control group were only treated with same amount of warm water. Neurological Severity Score (NSS) was evaluated after TBI at 0、3、5day respectively. All the rats were killed at the 5th day, the pathological change and brain water content were tested.
Results:1) Fluid percussing TBI model with the pressure 2.0-2.2atm is stable and reliable.2) The NSS had no significant differences between the YB treatment group and model control group at 0d(0> 0.05)。But compared with model control group, the NSS in YB treatment group greatly decreased at 3,5d (P<0.05).3) The pathological change of edema in the YB treatment group was better than the model control group. Compared with model control group, the brain water content in YB treatment group greatly decreased (P<0.05).
Conclusion:1) Fluid percussing TBI model with the pressure 2.0-2.2atm is suitable for this study.2) Yunnan Baiyao can reduce the NSS and brain water content of the TBI model rats, which may have therapeutic effect on TBI.
Part Two:The effects of Yunnan Baiyao on Caspase-3 expression and neurons apoptosis in Rats after Traumatic Brain Injury.
Objective:To explore effects of Yunnan Baiyao on Caspase-3 expression and neurons apoptosis in Rats after TBI.
Methods:42 SD rats were randomly assigned into seven groups:control group, 24h、72h、120h model control group and 24h、72h、120h Yunnan Baiyao(YB) treatment group. Each group contained 6 rats. Model control group and Yunnan Baiyao(YB) treatment group were established moderate or severe TBI model according to fluid percussing method. The YB treatment group was treated with Yunnan Baiyao (100mg/kg,q.d) and warm water, the control group and model control group were treated with same amount of warm water. Brains of rats were collected at different time (24h、72h、120h) after perfusion, then we determined the change of Caspase-3 expression and neurons apoptosis in tissues samples of frontal cortex and district in cornu-ammonis by immunohistochemistry.
Results:1)The expression of Caspase-3 in YB treatment group was significant lower than that in model control group (P<0.05), and was significant higher than that in control group (P<0.05) at 24h、72h、120h.2)The apoptosis index(AI) in YB treatment group was significant lower than that in model control group (P<0.05), and was significant higher than that in control group (P<0.05) at 24h、72h、120h.
Conclusion:There are neuronal apoptosis and change of Caspase-3 expression after moderate or severe TBI. Yunnan Baiyao can significantly restrain the neuronal apoptosis and the expression of Caspase-3 gene, which might have therapeutic effects for treating brain injury rats.
第一部分大鼠中重度创伤性颅脑损伤模型的建立和云南白药对大鼠创伤性颅脑损伤后神经功能及脑含水量的影响
目的在建立稳定可靠的液压打击TBI模型基础上探讨云南白药对中重度颅脑损伤后大鼠神经功能损伤评分及脑水肿程度的影响。
方法24只SD大鼠随机分为空白对照组、模型对照组、云南白药治疗组,各8只。云南白药治疗组和模型对照组采用液压打击法,造成中重度TBI模型。云南白药治疗组给予云南白药灌胃(100mg/kg体重,造模前一天与造模后5天内每天1次),空白对照组和模型对照组只给予等量温开水灌胃。术后第0天,第3天,第5天进行神经损害程度评分(NSS),第5天处死大鼠,取脑组织标本进行病理学检查,并计算干湿重比。
结果1)2.0-2.2atm致伤压力下造成的大鼠中重度TBI模型稳定可靠,死亡率可控。2)云南白药治疗组NSS评分较模型对照组在第0天无明显差别(P>0.05),在第3天,第5天显著降低,统计结果有显著性差异(P<0.05)。3)第5天云南白药治疗组脑组织病理改变及细胞形态学改变较模型对照组轻,脑含水量明显低于模型对照组,统计结果显示有显著性差异(P<0.05)。结论1)选择2.0-2.2atm致伤压力下液压打击TBI模型稳定可靠,可满足本实验研究需要。2)云南白药可以减低TBI后大鼠NSS评分,减轻脑水肿程度,对TBI具有治疗作用。
第二部分云南白药对中重度创伤性颅脑损伤后神经细胞凋亡及相关基因Caspase-3表达的影响
目的探讨云南白药对中重度颅脑损伤后大鼠神经细胞凋亡及相关基因Caspase-3表达的影响。
方法42只SD大鼠随机分组,空白对照组6只,模型对照组与云南白药治疗组各分为24h、72h、120h组,每组6只。云南白药治疗组和模型对照组采用液压打击法,造成中重度TBI模型。云南白药治疗组给予云南白药灌胃(100mg/kg体重,造模前一天与造模后每天1次),空白对照组和模型对照组只给予等量温开水灌胃。术后24h、72h、120h处死大鼠,灌注取脑组织标本免疫组化法检测皮质区和海马区细胞凋亡指数及Caspase-3的表达。
结果1)云南白药治疗组细胞凋亡指数在24h、72h、120h均低于模型对照组,且具有显著性差异(P<0.05),并明显高于空白对照组(P<0.05)。2)云南白药治疗组Caspase-3阳性细胞数在24h、72h、120h均低于模型对照组,且具有显著性差异(P<0.05),也明显高于空白对照组(P<0.05)。
结论中重度TBI后存在神经细胞凋亡和凋亡相关蛋白Caspase-3表达增加。云南白药可显著抑制TBI后大鼠神经细胞凋亡及Caspase-3基因表达,可起到对TBI的治疗作用。
On the basis of stable and reliable fluid percussing traumatic brain injury(TBI) models, we studied the therapeutic effects of Yunnan Baiyao on the model rats by testing Neurological Severity Score(NSS),cerebral edema, apoptosis index and related Caspase-3 gene expression.
Part One:The foundation of moderate or severe TBI model and the effect of Yunnan Baiyao on NSS and cerebral edema in TBI model rats.
Objective:On the basis of stable and reliable fluid percussing traumatic brain injury(TBI) models, we studied the brain protective effect of Yunnan Baiyao on the model rats by testing Neurological Severity score(NSS) and cerebral edema.
Methods:24 SD rats were randomly assigned into three groups:control group, model control group and Yunnan Baiyao(YB) treatment group. Each group contained 8 rats. Model control group and Yunnan Baiyao(YB) treatment group were established moderate or severe TBI model according to fluid percussing method. The YB treatment group was treated with Yunnan Baiyao (100mg/kg,q.d) and warm water, the control group and model control group were only treated with same amount of warm water. Neurological Severity Score (NSS) was evaluated after TBI at 0、3、5day respectively. All the rats were killed at the 5th day, the pathological change and brain water content were tested.
Results:1) Fluid percussing TBI model with the pressure 2.0-2.2atm is stable and reliable.2) The NSS had no significant differences between the YB treatment group and model control group at 0d(0> 0.05)。But compared with model control group, the NSS in YB treatment group greatly decreased at 3,5d (P<0.05).3) The pathological change of edema in the YB treatment group was better than the model control group. Compared with model control group, the brain water content in YB treatment group greatly decreased (P<0.05).
Conclusion:1) Fluid percussing TBI model with the pressure 2.0-2.2atm is suitable for this study.2) Yunnan Baiyao can reduce the NSS and brain water content of the TBI model rats, which may have therapeutic effect on TBI.
Part Two:The effects of Yunnan Baiyao on Caspase-3 expression and neurons apoptosis in Rats after Traumatic Brain Injury.
Objective:To explore effects of Yunnan Baiyao on Caspase-3 expression and neurons apoptosis in Rats after TBI.
Methods:42 SD rats were randomly assigned into seven groups:control group, 24h、72h、120h model control group and 24h、72h、120h Yunnan Baiyao(YB) treatment group. Each group contained 6 rats. Model control group and Yunnan Baiyao(YB) treatment group were established moderate or severe TBI model according to fluid percussing method. The YB treatment group was treated with Yunnan Baiyao (100mg/kg,q.d) and warm water, the control group and model control group were treated with same amount of warm water. Brains of rats were collected at different time (24h、72h、120h) after perfusion, then we determined the change of Caspase-3 expression and neurons apoptosis in tissues samples of frontal cortex and district in cornu-ammonis by immunohistochemistry.
Results:1)The expression of Caspase-3 in YB treatment group was significant lower than that in model control group (P<0.05), and was significant higher than that in control group (P<0.05) at 24h、72h、120h.2)The apoptosis index(AI) in YB treatment group was significant lower than that in model control group (P<0.05), and was significant higher than that in control group (P<0.05) at 24h、72h、120h.
Conclusion:There are neuronal apoptosis and change of Caspase-3 expression after moderate or severe TBI. Yunnan Baiyao can significantly restrain the neuronal apoptosis and the expression of Caspase-3 gene, which might have therapeutic effects for treating brain injury rats.
引文
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