| Cerebrovascular disease is harmful to the health of middle-aged and older for a class of diseases. As the population ages, the incidence of cerebrovascular disease rate increased year by year trend, China is the world's high incidence of cerebrovascular disease among the countries. According to statistics:In our country, there are 2,000,000～1,600,000 new cases of ischemic-stroke every year. 80% of which have varying degrees of dysfunction, 50%～70% of which left over Varying degrees of cognitive impairment, about 25% of which develop into dementia. Each year which die from ischemic stroke in our country as many as 750,000 of the total stroke mortality 3/4, the disease has become a human health and the serious threat to quality of life. Finding effective means of treatment and to explain its mechanism is still the field of brain science research hot spots.1. Literature researchWe found that stroke has various etiological factors, complicated pathogenesis, sudden onset and different clinical manifestations through researched on the etiology and pathogenesis theory of stroke from ancient to modern literatures. Lots of experimental studies have showed the underlying mechanism of acupuncture in the treatment of cerebral ischemia. Ischemic stroke was mainly due to regional cerebral circulatory disturbance caused by mental retardation, physical movement dysfunction as the main clinical manifestations of common diseases. Ischemic brain tissue can be divided into severe ischemia without reperfusion necrosis of the center is located in the surrounding areas and low-perfusion of ischemic penumbra (ischemic penumbra, IP). With the extension of ischemic time, the gradual expansion of necrotic area, while the penumbra shrinking. Ischemic lesions in the brain areas, neuronal necrosis and apoptosis of co-existence of necrosis located in the central area of ischemia, apoptosis was seen in the ischemic penumbra, apoptosis may determine the final infarct volume. Therefore, as soon as possible ischemic penumbra function, resist apoptosis and rescue the damaged cells is the key to the treatment of cerebral ischemia.Apoptosis is activated by cellular genes, enzymes and signal transduction pathways regulating is a "waterfall" process in the active cell death process. Among them, in particular, JAK (Janus kinase)/STAT (signal transducer and activator of transcription) family of control plays a decisive role in this. The current study found that:apoptosis may be related to STAT1 and STAT3, but activation related to cell survival and repair may be associated with STAT5 activation.Acupuncture in the treatment of cerebral ischemic disease has a unique effect. Our research is based on the study acupuncture intervention in a rat model of cerebral ischemia in the brain signal transduction systems, explore the lesions the region that the signal transduction system STAT family members and their mRNA expression levels and related factors modulation mechanism, a better understanding of acupuncture treatment for cerebral ischemia, interfere with the internal mechanisms of apoptosis, have a certain reference value.2. Experiments researchPurposeTo observe the effects of electroacupuncture (EA) on the signal transduction system STAT family members in the marginal zone of focal cerebral ischemia in rats at different time courses so as to further explores its underlying mechanism in the treatment of cerebral ischemia.MethodsA total of 300 male SD rats were divided randomly into sham group, model group, EA group, AG490 group and AG490+EA group. According to different drawing materials time there was 2hours, 1day, 3days, 1week and 3weeks time point after cerebral ischemia in each group, with 12 cases in each time point. Heat-coagulation-induced occlusion of the middle cerebral artery was performed to establish the model of focal cerebral ischemia in model group, EA group, AG490 group and AG490+EA group, but the middle cerebral arteries were exposed without occlusion in sham group. EA was immediately applied to Baihui (GV20) and Dazhui (GV14) for 30min in EA group and AG490+EA group. The treatment was given once daily. The sham group, model group and AG490 group did not receive acupuncture. Main outcome measures are following:(1) Neurological severty scores in every group.(2)The expression of neuronal apoptosis, in the marginal zone of focal cerebral ischemia in rats at different time points, was assayed with TUNEL staining method in different groups.(3) In the marginal zone of focal cerebral ischemia in rats at different time points, STAT1, STAT3 and STAT5 were measured by using immunohistochemical method, in stiu Hybridization (INH) and western blotting test.Results:(1)There had significant differences in the neurological deficit scores of model group and sham group at all stages, which showed that the models were made successfully. Cerebral ischemia in rats when 1d by the rest of the group scores were neurologic deficits in high level. Model group with the highest score, respectively EA group, AG490 group, and group compared AG490+EA significantly difference significance (P<0.01), and then three groups between the difference were not statistically significant (P>0.05). Model 3d rats, nerve dysfunction, partly mitigated rats contralateral to the lesions sham group compared with dumping, there were significant differences still (P<0.01), and EA group, AG490 group and AG490+EA group rats for nerve function recover gradually, score significantly lower model group (P<0.01), but the difference between three groups still no statistically significant (P>0.05). Ischemia in rats after each 1w leds, neurologic deficits condition improved, scores are relatively 3d group drops, and between the two slots in groups, and their respective sham group with between comparing the difference in a statistically significant (P<0.01). Three weeks later, model group and EA group of neural defect scale, but continue to decline group compared with no obvious difference 1w leds, sham group compared with all had significant difference (P<0.01), model group, EA group, AG490 group and AG490+EA group were no significant differences between groups (P>0.05).(2) Ischemia after 2h model group, EA group, AG490 group, the apoptotic cells AG490+EA group that started to increase, and time compared sham group was significantly difference (P<0.01), but no between-group difference obvious (P>0.05). Ischemia one day each byte continued to rise, in model group, including the level was the highest, EA group, AG490 group and the AG490+ EA group were significantly lower than model group (P<0.01), but the differences between the three groups were not obvious (P>0.05). Three days after each in ischemia and begins to decline, EA group, AG490 group and AG490 +EA group were far below the time of the model group (P<0.01), three groups were in the lower level, no obvious difference is between groups (P>0.05). Three weeks after the group to ischemia, had already closed to normal, no obvious difference between groups (P>0.05).(3) The expression of STATs, in the marginal zone of focal cerebral ischemia in rats at different time points, were influenced by EA.①The time of the sham group were only a small amount of STAT1 expression. Ischemia after two hours the STAT1 of model group, EA group, AG490 group and the AG490+EA group was beginning to rise, and the content of time group compared sham group all had significant difference (P<0.01), but no between-group difference obvious (P>0.05). Ischemia after a day, the STAT1 in groups express all continued to rise, among them with model group and the expression level AG49 group STAT1 higher, EA group and AG490+EA group of STAT1 express obviously lower than the above differences between the two groups, are statistically significant (P<0.01). Three days after the STAT1 ischemic each expression level began to drop, EA group and AG490+EA group were lower than the model group and time AG490 group (P<0.01). One week after the ischemia each intervention group can still detect STAT1 expression, to ischemia STAT1 protein three weeks after each already close to normal, no obvious difference between groups (P>0.05).②The time of the sham group were only a very small number of p-STAT3 expression. After two hours, model group, EA group, AG490 group and AG490+EA group of p - STAT3 expression level was beginning to rise, compared with sham group of time all had significant difference (P<0.01), which model group and EA group were significantly higher in increased significantly, the time AG490 group, AG490+EA group (both to P<0.01). And the model group and EA group, AG490 group and AG490+EA group differences between groups were not obvious (P>0.05). Ischemia After One day, the group of p- STAT3 expression level were rising- peak, model group in the most significant, and EA group followed it, AG490 group and AG490+EA groups even reached the peak in the group, also in the lower level. Three days after each in ischemia expression level of STAT3 p- began to decline, EA group, AG490 group and AG490+EA group were significantly lower than model group (P<0.01), and three groups no obvious difference between groups (P>0.05). Prolonged decline in the week after ischemia, the content of p-STAT3 had already closed to sham group level, no obvious difference between the groups (P>0.05). After three weeks to ischemia, the content of p-STAT3 in each group had already returned to normal level, no obvious difference between groups (P>0.05).③In the experiment each time the sham group only detected a small amount of p-STAT5 protein expression. Ischemia 2h model group, EA group, AG490 group and AG490+EA group that saw a large p-STAT5 expression, including model group and EA group in higher expression level, AG490 group and AG490+EA group expression was a bit low, but the model group and EA group, AG490 group and AG490+EA group, respectively between local area group no obvious difference (P>0.05), And with the same section of the sham group had significant differences (all P<0.01). Ischemia one day of the intervention group, p-STAT5 protein expression was the same with ischemia two hours, and ischemia three days later the intervention group, p-STAT5 levels began to decline, in which higher levels of EA group, the model next highest, AG490 group and AG490+EA group was lower than the expression of more than two groups (all P<0.01), to three weeks after ischemia in each group p-STAT5 expression has been near normal levels, no significant difference between groups (all P>0.05).3. ConclusionIn summary:EA can improve neurological severty scores in rats with cerebral ischemia level, its function in ischemic brain injury may be related to the internal mechanism to improve cerebral ischemic lesions electrical system regulation of STAT have a certain relationship, inhibition the content of STAT1 and p-STAT3 expression, increased expression of p-STAT5. Through this way of their protection, formed the repair mechanisms, launched suppressing lesions area half dark take cell apoptosis, restore damaged brain cells function. Preliminary validation of the "acupuncture stimulation - signal conditioning- starting from the homeostatic regulation of cerebral ischemia and its key role of local self-repair mechanism," the scientific hypothesis, for the acupuncture treatment of ischemic cerebrovascular disease provides a scientific basis and theoretical guidance. |
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