| Stroke is a disease endangers the lives and health of the middle-aged and elderly in our country seriously.The disease with high morbidity,high morbidity and high mortality characteristics,but also a higher rate of disease recurrence. Each year 200 million people in China have a new stroke,150 million people died of stroke,the cerebral infarction accounted for stroke from 60 to 80%,Because of its high fatality rate and disability rate for the country and many families caused heavy economic burden and life, so it has become a hot topic of modern medical research.Numerous studies have demonstrated,Ischemic brain injury is a series of pathophysiological changes more complex links process, early mainly produce free radicals, brain edema, excitatory amino acids toxicity mainly; Give priority to with inflammation and neuronal apoptosis in the medium term; Late on nerve regeneration, glial cell proliferation and angiogenesis brain reconstruction performance.After cerebral ischemia, angiogenesis is one of the important pathophysiological process of cerebral microcirculation reconstruction. The degree of angiogenesis and neural functional recovery after stroke was positively correlated. Vascular endothelial growth factor(VEGF) is found that the most powerful angiogenesis factor. Studies show that cerebral ischemia and hypoxia can induce the expression of vascular endothelial growth factor and its receptor, application of exogenous VEGF can promote cerebral ischemia half dark with the formation of new blood vessels, and reduce the cerebral infarction volume; ring oxidase- 2(cox-2) is speed limit important enzymes in the process of arachidonic acid into prostaglandin. In recent years a large number of studies have shown that cox-2 is involved in one of the important factor of angiogenesis; Angiogenin(angiopoietins, Ang) is gens specific effects on vascular endothelial cell growth factor,with great relevance of vascular endothelial apoptosis, angiogenesis, late maturity, stability and reconstruction.In recent years, many studies have confirmed that the three have an important role in angiogenesis, is a new field of international in recent years, but the one of traditional Chinese medicine, acupuncture effect on promoting angiogenesis research is relatively small, especially about needle head of promoting angiogenesis research rarely reported.Objective:Based on the above analysis, so the research on the model of focal cerebral ischemia in rats, based on the modified line bolt prepared MCAO model, quasi born awake brain begin to understand, removing stasis and promoting new as the therapeutic principle, take the top temporal after slash and top temporal front slash, by observing the head against focal cerebral ischemia reperfusion rats brain tissue expression of VEGF and cox-2,Ang-1,so as to explore possible mechanisms of angiogenesis of the head acupuncture game focal cerebral ischemia-reperfusion injury in rats.Methods: 100 grade clean SD male rats were randomly divided into model group(According to ischemia-reperfusion points 6h,24 h,48h, 96 h four groups), Head needle group(According to ischemia-reperfusion points 6h,24 h,48h,96 h four groups, Plus electroacupuncture respectively), The control group(Anesthesia in rats after cut the neck skin,only separation CCA and IPA to PPA, not plug wire plug), normal control group(normal rats not processing,feed water to eat),Each group of 10. Groups of rats with Zea-Longa grading method, along with the corresponding phase assess animal neurologic deficits. Immunohistochemical SABC method were used respectively to detect factors at each time point after cerebral ischemia reperfusion Ⅷ related antigen(F Ⅷ R : Ag) expression;By RT-PCR technique to detect ischemic penumbra zone groups of rats had Ang-1m RNA expression level;Using immunohistochemical method to detect ischemic penumbra zone groups of rats had the expression of VEGF and ES level;Using enzyme-linked immunosorbent(ELISA) method, fluorescence quantitative PCR detection testing ischemic penumbra zone groups of rats had cox-2 expression level.Results:(1)The rats after focal cerebral ischemia reperfusion, head of the needle group and the control group rats showed nerve dysfunction, mainly has: cerebral infarcts on lateral forefoot adduction, shoulder-in,tail dangling left fore cannot unbend or close to the chest wall, to the right when walking in circles,dumping,decreased muscle tone and so on. Using the Zea-Longa grading methods,24 h needle group and the control group rats nerve functions are gradually restored after CI/RP,but head needle group restore better than the control group rats,the most significant difference in the 96h;(2)Using the immunohistochemical SABC method, the head needle group compared with model group, FⅧR:Ag in the expression of cerebral microvascular in 24 h after ischemia began to increase(P<0.05),and continues to rise;The difference of 96 h after cerebral ischemia has significant statistical significance(P<0.01), acupuncture therapy with the extension of time can make its continue to rise. Cerebral ischemic penumbra thus head needle can effectively promote the capillaries of the newborn, and has obvious accumulative accumulation effect;(3)Using immunohistochemical method,normal control group, control group of Ang-1 have expression of equivalent, compared with model group, the scalp needle Ang-1 set of each point in time are relatively increased, including cerebral ischemia reperfusion 48 h Ang-1m RNA content reaches a peak, compared with model group, the difference was statistically significant(P<0.01, P<0.05);(4)Using immunohistochemical method to detect VEGF and ES, equivalent expression VEGF in normal control group and sham group, compared with model group, the scalp needle group rats at each time point tissue VEGF levels increase, ES content decreased, the brain ischemia reperfusion 48 h VEGF levels up to a peak, 24 h ES content of the resources,compared with model group,the difference was statistically significant(P<0.01, P<0.05);(5)Using enzyme-linked immunosorbent(ELISA)method, fluorescence quantitative PCR detection, the normal control group,control group of cox-2 small amounts of expression,the model group,the scalp needle group rats group at each time point cox-2 m RNA and protein content were reduced, the differences were statistically significant(P <0.05); the content of cox-2m RNA at 24 h, the content of cox-2 protein at 96 h in cerebral ischemia reperfusion injury were the most significant(P <0.01).Conclusion:Take the temporal after slash, temporal front slash, use head needle treatment of focal cerebral ischemia reperfusion rats,can increase the expression of FⅧR:Ag,VEGF,Ang-1,reduce the expression of ES 、cox-2,promote angiogenesis of cerebral ischemic area,reduce cerebral ischemic injury,have protective effect on ischemia brain tissue. |
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