| Objective:A rat model of ischemia-reperfusion was established to investigate whether the ele ctroa-cupuncture method of "tongdutiaoshen" could improve the neurological function o f isc-hemia-reperfusion rats by regulating the expression of autophagy responsibility-rel ated proteins P62,Beclin-1 and LAMP2.The mechanism of autophagy in ischemia-rep erfusion disease was observed by histophysiological staining and electron transmission techni-ques,which provided the theoretical basis and therapeutic target for the "tongdu tiaoshen" acupuncture therapy for the accumulation of clinical ischemia-reperfusion dise ase.Methods:60 SPF sprague-dawley rats were randomly divided into 30 models,15 in the bla nk group and 15 in the sham group.A modified Longa Artery Occlusion Reperfusion(MCAO/R)model was constructed to simulate Middle Cerebral Artery Occlusion(MC AO/R)in rats,and Reperfusion was performed after 2-h retrograde thrombectomy.In t he sham operation group,15 rats were successfully isolated and then suture without e mbolization.Zea Longa neurobehavioral score was performed after modeling to assess whet-her the modeling experiment was successful.The 30 rats with successful modelin g we-re divided into model group(n=15)and electroacupuncture group(n=15).The sec ond day after modeling,electroacupuncture acupuncture intervention began,electroacup unctu-re group(n=15)was treated with electroacupuncture,frequency of electroacupunc ture is 1/20 hz,density wave,30min/ time,1 time per day,a total of 7 days of interv ention,blank group(n=15),sham operation group(n=15),model group(n=15)only th e sa-me conditions for grabbing daily without other intervention.The neurological defici ts an-d improvements were assessed by Zea-longa5 scale on day 1 and day 7 of the i nterv-ention.After the intervention,TTC(triphenyl tetrazolium chloride)staining was u sed to observe the effects of electricity on the ischemic side tissue shape and cerebral infa-rction area of ischemia-reperfusion rats.The morphological and structural changes of nerve cells in the ischemic cortex of rats after electroacupuncture intervention wer e ob-served by transmission electron microscopy.The expressions of autophagy-related prot-einsP62,beclin-1 and LAMP2 in the ischemic cortex of rats in each group were detectedby Western Blot.Results:1.The results of neurobehavioral score showed that the blank group and the sha m oper-ation group had no symptoms of neurological defects,and both were 0 points.Comp-ared with the blank group and the sham group,the neurological function score of the model group and the electroacupuncture group significantly increased after mo deling(P<0.05).After 7 days of intervention,the neurological function score of the m odel gr-oup was significantly higher than that of the blank group and the sham group(P<0.05).Compared with the model group,the neurological function score of electroa cupunct-ure group was significantly reduced(P<0.05).The results showed that the met hod of "tongdutiaoshen" acupuncture could effectively improve the neurological defect of ischemia-reperfusion rats.2.The results of cerebral infarction area showed that there were no ischemic infar ction lesions in the blank group and the sham operation group,and there was no sign ifica-nt difference between the groups after model construction(P>0.05).After model constr-uction,compared with the blank group and the sham operation group,the mode l grou-p and the electroacupuncture group had significant ischemic infarction foci(P<0.05).Compared with the model group,the infarcted area of the electroacupuncture grou p was significantly reduced(P<0.05).3.Ultrastructure of the ischemia-side cortex was observed by transmission electron micr-oscopy: the morphological structure of nerve cells in the ischemia-side cortex of the i-schemia-reperfusion rats was observed structurally.Compared with the blank gro up an-d the sham group,the nerve cells in the model group were blurred,the cell me mbra-ne was damaged,the mitochondria were swollen,and even appeared to be vacuo us.C-ompared with the model group,the nerve cells in the electroacupuncture group had aclearer contour and a small amount of autophagosome formation was observed in the cells,suggesting that autophagy regulation may play an important role in the imp rovement of brain injury.4.Western Blot results showed that the expression of beclin-1 protein was signific antly increased in both the model group and the electroacupuncture group compared w ith the blank group and the sham group(P<0.05),and the difference was statistically sign-ificant(P<0.05).Compared with the blank group and the sham group,the expre ssion of P62 protein in the model increased,and that in the electroacupuncture group decrea-sed,with statistically significant differences(P<0.05).Compared with the model group,the expression of electroacupuncture histone decreased,and the difference was statisti-cally significant(P<0.05).Compared with the blank group and the sham group,LAMP2 protein expression in the model decreased,and that in the electroacupuncture groupincreased,with statistically significant differences(P<0.05).Compared with the model group,the expression of electroacupuncture histone increased,and the difference was s-tatistically significant(P<0.05).The results showed that electroacupuncture inter ventionin ischemia-reperfusion rats to improve the neurological function of rats was cl osely related to the regulation of autophagy-related protein expression.Conclusion:The method of "tong du tiao shen" acupunctur can improve the range of cerebral infarction in rats with cerebral ischemia/reperfusion injury,positively regulate the ultr astructure of cerebral nerve cells,regulate the expression levels of autophagy-related pr oteins P62,Beclin 1 and LAMP2 a,and thus protect the cerebral nerve function,whic h may be one of the mechanisms by which acupuncture improves the nerve function of MCAO/R rats. |
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