The Protection Of Danxingtongluotang(DX) On Cerebral Iachemia-reperfution

Objective: The modern medical research thought that the cerebral ischemia- reperfusion injury is an important pathophsiology mechanism that causes the various cerebral disease,and apoptosis is one of the important mechanisms of the cell death in the process of the organ and the organization ischemia-reperfution. Therefore the intervention apoptosis as to prevent and controlischemia-reperfusion injury is already to arouse the demestic and foreign widespread interests again . In the different time after the ischemia,there are many kinds of reactivitigene successively to express. Among these,p53gene, as a transcription factor,promote cell's apoptosis through activating a series of downrivers gene to press. Caspase–3 expression leads the cerebral ischemia tardive nurve cell death and the nurve cell apoptosis.Danxingtongluo(DXTL)is a mixture of some herbs,such as Danshen, Dannanxing, Sanqi,etc.They have functions of cleaning away stagnated blood and heat,restoring normal functioning of body to consolidated the constitution and resolving mass.In clinical we can get a fine therapeutic effet by using it to the prognosis of ischmia cerebrovascular disease.In the prophase experiment,it is indicated that DXTL can depress the neurocyte aptosis of ischemia-reperfusion rat by reducing ischemia-reperfusion model rat NO contents,increasing SOD and evaluating the expression of gene bcl-2.The purpose of this study was to observe cerebral ischemia-reperfusion model rat P53 and caspase-3 gene expression and observe serum LDH contents in the brain,and discuss the protective functions and further to explor the mechanism of DXTL.Methods:forty healthy male Wistar rats were randomly divided into 5 groups, 8 rats per group: sham operation group , ischemia/reperfusion (I/R) group, DXTLD high dosage group, DXTLD low dosage group and Nimodipine(NIMO) group. DXTLD groups were given to DXTLD at dose of 15ml/kg,7.5ml/kg;NIMO at10ml/kg,once a day. Saline were given to sham operation group and I/R group at 7.5ml/kg weight. After 7-days of intragastricd, the rats of all the groups except sham operation group were made into cerebral ischemia reperfusion injury models by thread embolism method. All the rats were killed and their brains were taken out after 2h ischemia and 24h reperfusion injury. The HSP70 expression and Caspase-3 expression in brain tissue were measured with immunohistochemical method .HE dyeing observe the histomorphology change in rats of each group. Photocolorimetry measure the content of LDH in serum.Results:1.histomorphology observation: groupA(sham-operated):normal cellular structure.group B(the cerebral ischemia-reperfusion model): the cellula nervosa struvture are fuzzy,cell body swelling,the Nissl's body decreased or vanished, varying degree the nucleus anachromasis, karyopyknosis and karyolysis, nuclear body is irregular, has obviously soften focus of infection, the peripheral obviously neutrocyte infiltrates. In the treatment groups and nimmodipine group the neurocyte arrangement lightly disorder. And we can also see the nucleus anachromasis, karyopyknosis and karyolysis,but they were decreased compared with the model group .The group C and D were the most significant.2.Effect of DXTL on expression of caspase-3 and p53: the group A can be seen caspase-3 positive cell and proteinum expressed weakly;after the cerebrain ischemia (MCAO)2 hours followed 24 hours reperfusion ,caspase-3 and p53 positive cell were increased remarkably in group B.Compared with model group ,the treatment group were decreased significantly .The group C and D were decreased compared with the group E. But the C group had no significant with difference with the group D.3.Seral LDH level of model group were increased significantly compared with sham-operation group (P<0.05);the treatment group were increased compared with the sham-operation group,but it decreased remakably compared with the model group(P<0.05) .Conclusion: Danxingtongluotang has the obvious protection against ischemia- reperfusion injury the effect may be related to decreasing the seral LDH contents, depress the apoptosis by reducing the expressive levels of caspase-3 and p53 positive cell .