Effects Of Modified Buyanghuanwu Decoction On Expression Of BDNF And NGF In Rat Brain After Ischemia/reperfusion

ObjectiveIschemic cerebral vascular disease,considered as "stroke"in Traditional Chinese Medicine,is one of the top 3 disease which has severe harm to human health,and has high incidence,mortality and disability rate.Modern medical research has convinced that series of cascade reactions will be triggered after ICVD.As excitatory amino acid toxicity,oxygen free radical injury,cell apoptosis,inflammation,et al.Which will cause neural cells injury.BDNF,NGF can repair injured neurons,maintain the function of central and peripheral nervous system.Therefore,the increased BDNF,NGF concentration of cerebral ischemia can promote the recovery of neural function and remission of clinical symptoms.Traditional Chinese Medicine preventing stroke has long history,the medical classic of the yellow emperon,2000 years ago had record about stroke.The buyanghuanwu decoction is the classical prescription for stroke first created by Wang Qingren in Qing dynasty,and recored in Yilingaicuo.The main function is upplementing qi and activating blood circulation and dredging meridian,which has obvious effect in treating stroke.The experiment observe the BDNF and NGFconcentration change after using Modified Buyanghuanwu decoction in focal cerebral ischemic/reperfusion rats,and explore the neuron protection mechanism of ischemic cerebral injury.Methods and Materials1.Institution of animal modelsSeventy clean level healthy male SD rats,randomly devided into sham operation group,model group and treating group.The last two group then be devided into three subgroups which are 12h,24h and 48h after ischemia respectively,10 rats in each group.Preparation focal cerebral ischemic model by suture method occlusing the left middle cerebral artery.The conditions sham operation is the same to other two groups besides not occlusing by thread.2.Brain tissue sliceAfter successfully constracting the model,rats are sacrificed at different time point.The brain behind optic chiasma is taken out,and then situated in 4℃,4% paraformaldehyde prosphate buffer solution for 24h.Regular dehydration,waxing, embedding and pressing into 7μm thick coronal slice.Then placing on the glass slide and preserving it in 60℃temperature box for 2 hours.3.Immunohistochemistry stainingUsing SABC method,and immunohistochemistry techonology,observing the immunoreactive cell of BDNF and NGF,statisticalling the number of immunoreactive cell with image analysis software.4. Analysis of the dataThe result is displayed as Mean±standard devition, P< 0.05 is considered as statistically significant by t-test.ResultsCompared with the sham-operated group, the expression of BDNF and NGF increased at 12hours,24hours and 48hours after ischemia(P< 0.01). Compared with the ischemia group, the expression of BDNF increased at different stages after ischemia(P < 0.01). The expression of NGF is also(P< 0.05), and reached the climax after 48 hours.ConclusionsThe BDNF and NGF has increased in focal cerebral ischemia/reperfusion rats after using Modified Buyanghuanwu Decoction,promoting the recovery of injuried neurons.