Influence Of Buyang Huanwu Decoction On Neurologic Protection And Expression Of Akt In Ischemia-reperfusion Rats

ObjectiveCerebrovascular disease is common and the mortality and morbidity are very high. Ischemic cerebrovascular disease account for about 70% to 80%. The pathophysiology of ischemic brain injury is very complicated. Platelet dysfunction is one of the most important inducing factors of ischemic cerebrovascular disease. Buyang Huanwu Decoction is a classical prescription in the treatment of ischemic cerebrovascular disease. Phosphatidyl inositol 3 kinase/protein kinase B(PI3K/Akt) signal transduction pathway is an important cellular pathway in the body and it is closely related to platelets activation.This study is based on rats focal cerebral ischemia model. Through platelet activating factor(PAF), CD40 L, CD62 P detection, discussing the effects of Buyang Huanwu decoction on the activity of platelets. By the research of Akt protein expression of PI3K/Akt signal pathway, exploring the mechanism of Buyang Huanwu Decoction inhibiting ischemic injury. Providing new ideas for the treatment of ischemic cerebrovascular disease with Chinese medicine and scientific basis for clinical medication. Methods 1.Animal experimentThe 100 specific pathogen free SD male rats were randomly divided into 5 groups: sham operation group, model group, positive control group(6.8mg·kg-1) and Buyang Huanwu Decoction(high, low dose)group(26,13g·kg-1). Each group contained 20 rats,The sham operation group and model group were given distilled water.Using modified nylon embolization method to duplicate middle cerebral artery occlusion(MCAO) reperfusionrat rats model after 14 days’ administration. For 1 hour of cerebral ischemia and 6 hours of reperfusion, detecting the scores of nerve function. 2. Testing items and methodsUsing TTC staining method to detect the area of cerebral infarction; using HE staining to observe neurons changes of hippocampus CA1 and cortex area;using WST-1 method to detect brain tissue SOD activity, using TBA method to detect the MDA contents in serum; using Elisa method to detect the content changes of PAF, CD40 L and CD62 P in serum;using IHC method to detect the protein expression of p- Akt(Ser473) and Akt(Ser473) in brain hippocampus;using WB method to detect the protein expression of p-Akt and Akt in hippocampus. ResultsMCAO rat models were prepared successfully. High and low dose of Buyang Huanwu Decoction group could reduce the nerve function score of MCAO reperfusion rats, reduce the cerebral infarction area, improve the area of pathological damage in hippocampus CA1 and cortex area, improve the structure of the nerve cells and nucleus pycnosis situation, relieve the damage of nervous system, elevate total SOD activity in brain tissue, reduce the MDA content in serum, improve the free radical balance, lower the PAF, CD40 L, CD62 p levels in serum, inhibit the activation of platelets; increase the content of p-Akt protein in brain tissue, and activate the PI3K/Akt signal pathway. Buyang Huanwu Decoction in high dose had better effects. ConclusionsBuyang Huanwu Decoction pretreatment can inhibit platelet activation, raise the expression of p-Akt effectively, It prompts that Buyang Huanwu Decoction has certain protective effect on brain tissues of MCAO reperfusion rats. The mechanism may be related to the inhibition of platelet activation and the regulation of PI3K/Akt signal pathway.