Stroke is one of three important diseases threaten healthy of human being. Ischemia stroke with high mortality and disability rate is accout for 55%-80% of stroke. Cerebral ischemia reperfusion injury is one mainly pa...Stroke is one of three important diseases threaten healthy of human being. Ischemia stroke with high mortality and disability rate is accout for 55%-80% of stroke. Cerebral ischemia reperfusion injury is one mainly pathological mechanism of ischemia stroke. The functional disturbance of the network of neuro-endocrine and immune (NEI) induced by cerebral ischemia reperfusion heighten abnormally glucocorticiod (GCS) in hippocampus and plasm ,and aggravate pathological impairment after cerebral ischemia reperfusion. Recently investigation showed that excessive GCS is one of important pathological factors causing neuron apoptosis in major brain regions such as hippocampus related to cognitive function, which is responsible for vascular dementia (VaD) after stroke. At present, the mechanism induced neuron apoptosis by higher GCS is remained unkown. JianNaoAn with function of BuShenHuoXueHuaTan, is one effective compound of Chinese herbal drug on curing VaD. The proceding experimental result has confirmed the function of JianNaoAn down-modulated genes expression of IL-1 ,CRF related to NEI and relieved impairment of nerve cells induced by cerebral ischemia -reperfusion. We investigate the action about excessive GCS causing neurons apoptosis after cerebral ischemia and the influence of JianNaoAn on this pathological machenism through radioimmunoassay (RIA), immunohistochemistry, in situ hybridization and TUNEL experimental techniques. Furthermore the protective function of JianNaoAn on neuron after cerebral ischemia-reperfusion and potentially pharmacological methanism to prevent and cure post-stroke dementia were revealed. At 2,6,12,24 h of different ischemia-reperfusion points, the concentration of cortisiol of the JianNaoAn treated group in plasm and hippocampus decreased significantly compared with the model group (P<0.05). The number of positive cells is reduced significantly compared with the model group by TUNEL staining method, especially in reperfusion 12 h,the distinction in number of positive cell between the JiaNaoAn treated group and the model group is very significant (P <0.01) . The proteins and mRNA expression of NMDAR,cytochrome C and caspase 3, which are the key genes in the apoptosis pathway of depended mitochondrion, are down-modulated in the treatment group and the antagonist group ,and there were distinct difference compared with the model group. However,there is no significant difference about the expression of glucocorticoid receptor between the treatment group and the model group. By the apoptosis pathway of dependent mitochondrion that excessive GCS caused neurons apoptosis in hippocampus is confirmed in our experiment. But JianNaoAn can reduce selectively GCS and inhibit the activity of the apoptosis pathway of dependent mitochondrion induced by excessive GCS, and then decrease the degree of apoptosis in hippocampus after cerebral ischemia- reperfusion. The protective action is one function of preventing neurons from injury resulted from cerebral ischemia-reperfusion. Meanwhile, the function of BuShenHuoXueHuaTan method on regulating the disturbance of network of NEI induced by cerebral ischemia-reperfusion may be an important biological mechanism protecting neurons, preventing and treating vascular dementia.展开更多
文摘Stroke is one of three important diseases threaten healthy of human being. Ischemia stroke with high mortality and disability rate is accout for 55%-80% of stroke. Cerebral ischemia reperfusion injury is one mainly pathological mechanism of ischemia stroke. The functional disturbance of the network of neuro-endocrine and immune (NEI) induced by cerebral ischemia reperfusion heighten abnormally glucocorticiod (GCS) in hippocampus and plasm ,and aggravate pathological impairment after cerebral ischemia reperfusion. Recently investigation showed that excessive GCS is one of important pathological factors causing neuron apoptosis in major brain regions such as hippocampus related to cognitive function, which is responsible for vascular dementia (VaD) after stroke. At present, the mechanism induced neuron apoptosis by higher GCS is remained unkown. JianNaoAn with function of BuShenHuoXueHuaTan, is one effective compound of Chinese herbal drug on curing VaD. The proceding experimental result has confirmed the function of JianNaoAn down-modulated genes expression of IL-1 ,CRF related to NEI and relieved impairment of nerve cells induced by cerebral ischemia -reperfusion. We investigate the action about excessive GCS causing neurons apoptosis after cerebral ischemia and the influence of JianNaoAn on this pathological machenism through radioimmunoassay (RIA), immunohistochemistry, in situ hybridization and TUNEL experimental techniques. Furthermore the protective function of JianNaoAn on neuron after cerebral ischemia-reperfusion and potentially pharmacological methanism to prevent and cure post-stroke dementia were revealed. At 2,6,12,24 h of different ischemia-reperfusion points, the concentration of cortisiol of the JianNaoAn treated group in plasm and hippocampus decreased significantly compared with the model group (P<0.05). The number of positive cells is reduced significantly compared with the model group by TUNEL staining method, especially in reperfusion 12 h,the distinction in number of positive cell between the JiaNaoAn treated group and the model group is very significant (P <0.01) . The proteins and mRNA expression of NMDAR,cytochrome C and caspase 3, which are the key genes in the apoptosis pathway of depended mitochondrion, are down-modulated in the treatment group and the antagonist group ,and there were distinct difference compared with the model group. However,there is no significant difference about the expression of glucocorticoid receptor between the treatment group and the model group. By the apoptosis pathway of dependent mitochondrion that excessive GCS caused neurons apoptosis in hippocampus is confirmed in our experiment. But JianNaoAn can reduce selectively GCS and inhibit the activity of the apoptosis pathway of dependent mitochondrion induced by excessive GCS, and then decrease the degree of apoptosis in hippocampus after cerebral ischemia- reperfusion. The protective action is one function of preventing neurons from injury resulted from cerebral ischemia-reperfusion. Meanwhile, the function of BuShenHuoXueHuaTan method on regulating the disturbance of network of NEI induced by cerebral ischemia-reperfusion may be an important biological mechanism protecting neurons, preventing and treating vascular dementia.
