The Mechanism Research Of Acupuncture Treatment For Ischemic Stroke Based On AMPK Levels

Objective:To establish a rat model of middle cerebral artery occlusion(MCAO),investigating the AMPK-based molecular mechanism of acupuncture intervention on ischemic stroke,and assessing the effects of acupuncture on intracerebral energy metabolism and the pathogenic significance in the disease of ischemic stroke,all in an effort to provide a scientific foundation for clinical application.Methods:1.Protective effect of acupuncture on ischemic stroke(1)Male SPF grade Sprague-Dawley rats that were 2-3 months old with healthy bodies were used to create the ischemic stroke model.(2)Rats that had successfully undergone surgery were divided into two groups,one receiving acupuncture and the other receiving a model,in accordance with a random number table.There were 10 rats in each group,and the rats in the sham operation group underwent the exact same procedure but with no suture plug.(3)The longa score was used to gauge the neurological health of the rats in each group.(4)Using hematoxylin eosin staining(HE)to observe the neuropathological and neuropathological changes in the brain of ischemic rats.(5)ATP levels were measured by enzyme-linked immunosorbent assay(ELISA).2.Mechanism of acupuncture on ischemic stroke injury(1)Using immunohistochemistry assay to detect the expression of AMPK and p-AMPK in brain tissue;(2)Western blot was used to detect the protein expression levels of AMPK and p-AMPK in brain tissue;(3)The real-time reverse transcription polymerase chain reaction(RT-qPCR)were used to detect the expression of AMPK mRNA in brain tissue.Results:1.Protective effect of acupuncture on ischemic stroke(1)Rats with middle cerebral artery occlusion showed contralateral neurological impairment.(2)The experimental results of modified Zea-Longa showed that the neurological scores of rats were significantly increased after modeling surgery.After the intervention,there was no significant change in the neurological function score of the model group.And the neurological function score of acupuncture group was significantly lower than before treatmentand model group.(3)HE staining showed:in the brain tissue of the sham operation group,the cytoplasm of nerve cells was abundant,the nucleus was large and round,and no histopathological changes were observed.Meningeal edema,pyknosis and hyperchromatism of nerve cell nuclei,ballooning degeneration,cytoplasmic vacuolation and loose arrangement of connective tissues could be seen in the brain tissue in the ischemic area of the model group.After acupuncture intervention,necrotic neurons were still observed in the ischemic area,showing a significant increase in fibroblasts.Compared with the model group,the number of cells with normal and complete cytoplasm and nuclear structure was significantly increased,and the cells were arranged in a relatively neat and orderly manner.(4)ELISA detection shows:Compared with the model group,acupuncture can significantly increase the level of ATP expression in the brain tissue of MCAO rats(P<0.05).2.Mechanism of acupuncture on ischemic stroke injury(1)Immunohistochemical results showed that in the brain tissue of rats in the sham operation group,a small amount of AMPK and p-AMPK expressions were primarily found in the cytoplasm,with some being present in the nucleus.Granules that were brownish yellow represented the favorable expression.Rats in the model group had higher levels of positive AMPK expression in their brain tissue compared to the control group,but this difference was not statistically significant(P>0.05),and p-AMPK was significantly increased(P<0.01).The acupuncture group’s AMPK level in its brain tissue were higher(P<0.05)than those of the model group and the sham surgery group,and its expression of p-AMPK was significantly higher(P<0.01)as well.(2)Results for the Western Blot:compared with the sham surgery group,the relative expression level of AMPK protein in the brain tissue of the model group had no significant change(P>0.05),while the level of p-AMPK protein was significantly increased(P<0.01);the relative expression levels of AMPK and p-AMPK proteins in the brain tissue of rats in the acupuncture group were significantly higher than those in the model group(P<0.01)(3)According to RT-qPCR findings,rats in the model group had somewhat higher levels of AMPK expression in their brain tissue compared to the sham acupuncture group(P<0.05),while rats in the acupuncture group had significantly higher levels of AMPK mRNA expression compared to the model group(P<0.01).Conclusions:Acupuncture can effectively alleviate the neurological impairment caused by cerebral ischemia and reduce the tissue damage of nerve cells.Its effect may be achieved by increasing the phosphorylation level of AMPK in the brain,promoting the expression of its protein and mRNA,enhancing brain tissue’s energy metabolism.