| Preface:Nowdays the ischemia cerebral vascular disease has threatened the health of human being. The ischemia vascular disease has the character of high morbidity, high mortality rate and high cripples rate. Chinese medicine has long history of preventing and controlling stroke. The Chinese medicine classic work《the yellow Emperor Canon of Internal Medicine》has the record of cause, treatment principle and Prevention health care for stroke. Especially acupuncture were used to treat stroke. Recently Clinical researches demonstrate that puncturing the skull point can improve moving ability of the paralytic limb and make the scale of the patient neurology dysfunction better. The MCAO rats were adopted in this research and the technology of TUNEL,RT-PCR,ELISA,immunohistochemistry and electron-microscope were used. the preventing effect of puncturing the head point were observed systematically and the mechanism of the acupuncture anti cerebral ischemia were probed through the cell apoptosis, the express of the associated apoptotic gene, inflammation reaction and neuronal micro structure. This research had provided the scientific evidence for the head needle treating the ischemia cerebral vascular disease clinically.Materials and Methods:The health rats were randomly divided into 3 groups. A group(sham-operated group);B group(ischemia/reperfusion group);C group (acupuncture treatment group). The middle cerebral artery occlulsion(MCAO) were established by using the longas method. The infarct volume were studied by TTC staining after 24 hours of ischemia and reperfusion, scores of neurological functions deficits were evaluated after 3h,24h,72h following ischemia /reperfusion, neuronal apoptosis were detected by the TUNEL staining, the expression of bcl-2 and bax were observed by immunohistochemistry after 3h,24h,72h following ischemia /reperfusion, the IL-1βwas detected by using the ELISA method, the expression of caspase-3mRNA,caspase-8mRNA were detected by RT-PCR methods, the neural micro structure was observed by electron microscope. All data were expressed as mean±SD. Statistical analysis were carried out by spss10.0.Results:1,Infarct volumesThere were not infarcts volume in A group. The infarcts in B,C group were mainly in frontal-parietal cortex, caudates nucleus and putamen nucleus. The infarcts volumes of C group were significantly less than B group(P<0.05)2,Evaluation of neurological functions deficits Scores of neurological functions deficits of C group were less than that of B group after 3h,24h,72h following ischemia/reperfusion. Compared with the Scores of neurological functions of B group, the difference between C group and B group is significant after 24h,72h following ischemia/reperfusion.3,Neuronal apoptosisApoptotic neurons were mainly located around ischemic tissues, that was ischemic penumbra. There were few apoptotic neuronal cells in A group. The numbers of apoptotic cell increased after 3h following cerebral ischemia/reperfusion, then gradually increased as time went on, at 24h reached the peak, then decreased gradually. The number of apoptotic neuronal cells in A group were different from them in both B and C group significantly. The number of apoptotic neuronal cells in C group is significant less than that in C group(p<0.01,P<0.05)4,ImmunohistochemistryThere were a few expression in A group. The expression of B and C group were mainly located in ischemic penumbra. The expression of bcl-2 and bax began to increase after 3h following cerebral ischemia/reperfusion, reached the peak at 24h, the peak of bcl-2 continuing to 72h following cerebral ischemia/reperfusion, the expression of bax began to decrease after 72h following cerebral ischemia. The expression of bcl-2 in C group is significantly enhanced than that in B group, the expression of bax in C group is weaker than that in B group.5,ElisaThere were a few IL-1βin A group. IL-1βbegan to increase after 3h following cerebral ischemia/reperfusion, reached the peak at 72h. the number of IL-1βin C group is less than that in B group significantly.6,Reverse transcription-polymerase chain reaction (RT-PCR)There were a little expression of caspase-3mRNA and caspase-8 mRNA. The expression of caspase-3mRNA and caspase-8 mRNA gradually increased as time went on following ischemia/reperfusion, reached the peak at 24h following cerebral ischemia/reperfusion. The expression of caspase-3mRNA and caspase-8 mRNA in D group is less than that in B group significantly.7,Electron-microscopeThe neurons,gliocyte and structure of capillaries were in integrity in A group. There were a lot of vacuole in neuron and organelle disappeared in B group. There were also a lot of vacuole in gliocyte and the cavity of capillaries became smaller, vascular endotheliocyte became bigger in B group. In C group there were a few vacuole and a lot of organelle such as endoplasmic reticulum,mitochondria,ribosomes, nucleus was big and chromatin was uniform in neuron; endoplasmic reticulum was expansion mildly, the number of golgi body became more in gliocyte; the cavity of capillaries became bigger and endotheliocyte became smooth.Conclusion:1,Acupuncturing the head point-line of anterior oblique line of vertex-teporal and posterior oblique line of vertex-teporal can improve the scores of neurological functions deficits, decrease the infarct volume and make the structure of neuron keeping integrity significantly. this result indicate that acupuncturing the head point have protective effect on cerebral ischemia/reperfusion.2,Acupuncturing the head point has the protective effect on cerebral ischemia/reperfusion by decreasing neuronal apoptosis in ischemia penumbra.3,Acupuncturing the head point can significantly increase bcl-2 and decrease bax 4,Acupuncturing the head point can reduce inflammation reaction, lower IL-1βof ischemic penumbra.5,Acupuncturing the head point can significantly decrease the expressions of caspase-3mRNA and caspase-8 mRNA following cerebral ischemia/reperfusion and inhibit the neuronal apoptosis.
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