黄芪加红花对大鼠脑缺血再灌注后神经细胞凋亡的影响
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摘要
目的拟观察黄芪注射液和红花注射液对SD大鼠局灶性脑缺血再灌注模型脑细胞凋亡、天冬氨酸特异性半胱氨酸蛋白酶-8(Caspase-8)和天冬氨酸特异性半胱氨酸蛋白-3(Caspase-3)表达的影响,来探讨黄芪注射液和红花注射液治疗脑缺血性中风的作用机理。
方法将SD雄性大鼠随机分为假手术组、模型组、红花组、黄芪组、黄芪加红花组,各药物组腹腔注射相应药物,假手术组与模型组腹腔注射等量生理盐水;采用线栓法制备局灶性脑缺血再灌注模型。分别在缺血再灌注12h,24h,48h对其神经系统体征进行客观评分后断头处死。行HE染色观察各时间点脑组织病理学形态的改变;并运用TUNEL法观察缺血再灌24h凋亡的情况:免疫组化的方法检测Caspase-8,Caspase-3的表达,在光镜下分别计数其阳性细胞数。
结果与模型组比较,黄芪加红花组、红花组和黄芪组均能改善脑缺血再灌注模型鼠神经损伤症状;减轻脑缺血再灌注时的病理损伤;减少凋亡阳性细胞的表达;抑制Caspase-8、Caspase-3阳性细胞在各时间点的表达(p＜0.05),且黄芪加红花组与红花组、黄芪组比较,各时间点Caspase-8、Caspase-3阳性细胞数减少更明显,差异有显著性(p＜0.05)。
结论黄芪注射液协同红花注射液可促进脑缺血再灌注大鼠脑细胞凋亡的减少,其机制可能和抑制Caspase-8,Caspase-3的表达有关;黄芪注射液和红花注射液协同治疗效果优于单独运用。
Objective To study the therapeutic effects of radix astragali injection(RA) and safflor injection(SI) on Cysteinyl aspartate specific protease-3(Caspase-3),Cysteinyl aspartate specific protease-8(Caspase-8) and apoptosis in rats brains following local cerebral ischemia/ reperfusion.
Methods Male adult SD rats were randomly divided into five groups:the sham-operated group, the model group,the SI group,the RA group and the RA+SI group.The model of middle cerebral artery occlusion(MCAO) was established by thread ligation method,and rats were injected intraperitoneally corresponding medicine in different groups at 12h,30min before operation and every other 12h after operation.In sham-operated group and the model group, equally dose saline was injected intraperitoneally.Neurological system symptoms were evaluated at 12h,24h,48h after reperfusion.Then the rats were decapitated at 12h,24h,and 48h after reperfusion,and rats brains were taken for histopathological,TUNEL and immunohistochemistry examinations.Positive reacted cells of apoptosis,Caspase-8 and Caspase-3 are counted under light microscope at different time points of ischemia/reperfusion.
Results The score of neurological system damage symptoms and the examination of histopathological,TUNEL,immunohistochemistry show that the cerebral ischemic damage in the RA group,the SI group and the RA+SI group was significantly milder than that in the model group(p＜0.05).Positive immune reacted cell amounts of Caspase-8 and Caspase-3 are alleviate in RA+SI group as compared with the RA group and the SI group in 12h,24h,48h ischemia/ reperfusion(p＜0.05).
Conclusions The cooperation protective effect of RA and SI injection therapy against cerebral ischemia reperfusion injury might be associated with alleviating the positive expressions of Caspase-8 and Caspase-3.Moreover,The cooperation therapeutic effect of RA and SI is superior than the unity therapeutic effect of RA or SI.
方法将SD雄性大鼠随机分为假手术组、模型组、红花组、黄芪组、黄芪加红花组,各药物组腹腔注射相应药物,假手术组与模型组腹腔注射等量生理盐水;采用线栓法制备局灶性脑缺血再灌注模型。分别在缺血再灌注12h,24h,48h对其神经系统体征进行客观评分后断头处死。行HE染色观察各时间点脑组织病理学形态的改变;并运用TUNEL法观察缺血再灌24h凋亡的情况:免疫组化的方法检测Caspase-8,Caspase-3的表达,在光镜下分别计数其阳性细胞数。
结果与模型组比较,黄芪加红花组、红花组和黄芪组均能改善脑缺血再灌注模型鼠神经损伤症状;减轻脑缺血再灌注时的病理损伤;减少凋亡阳性细胞的表达;抑制Caspase-8、Caspase-3阳性细胞在各时间点的表达(p＜0.05),且黄芪加红花组与红花组、黄芪组比较,各时间点Caspase-8、Caspase-3阳性细胞数减少更明显,差异有显著性(p＜0.05)。
结论黄芪注射液协同红花注射液可促进脑缺血再灌注大鼠脑细胞凋亡的减少,其机制可能和抑制Caspase-8,Caspase-3的表达有关;黄芪注射液和红花注射液协同治疗效果优于单独运用。
Objective To study the therapeutic effects of radix astragali injection(RA) and safflor injection(SI) on Cysteinyl aspartate specific protease-3(Caspase-3),Cysteinyl aspartate specific protease-8(Caspase-8) and apoptosis in rats brains following local cerebral ischemia/ reperfusion.
Methods Male adult SD rats were randomly divided into five groups:the sham-operated group, the model group,the SI group,the RA group and the RA+SI group.The model of middle cerebral artery occlusion(MCAO) was established by thread ligation method,and rats were injected intraperitoneally corresponding medicine in different groups at 12h,30min before operation and every other 12h after operation.In sham-operated group and the model group, equally dose saline was injected intraperitoneally.Neurological system symptoms were evaluated at 12h,24h,48h after reperfusion.Then the rats were decapitated at 12h,24h,and 48h after reperfusion,and rats brains were taken for histopathological,TUNEL and immunohistochemistry examinations.Positive reacted cells of apoptosis,Caspase-8 and Caspase-3 are counted under light microscope at different time points of ischemia/reperfusion.
Results The score of neurological system damage symptoms and the examination of histopathological,TUNEL,immunohistochemistry show that the cerebral ischemic damage in the RA group,the SI group and the RA+SI group was significantly milder than that in the model group(p＜0.05).Positive immune reacted cell amounts of Caspase-8 and Caspase-3 are alleviate in RA+SI group as compared with the RA group and the SI group in 12h,24h,48h ischemia/ reperfusion(p＜0.05).
Conclusions The cooperation protective effect of RA and SI injection therapy against cerebral ischemia reperfusion injury might be associated with alleviating the positive expressions of Caspase-8 and Caspase-3.Moreover,The cooperation therapeutic effect of RA and SI is superior than the unity therapeutic effect of RA or SI.
引文
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