| Objectiue:The aim of this study was to observe the effect of acupuncture on the content of Fas,Fas L in serum and caspase-3 and caspase-9 positive cells in brain tissue.and to explore the mechanism of acupuncture in alleviating cerebral ischemia / reperfusion injury.Methods:A total of 100 Sprague-dawley male rats,220g-300 g were randomized into three groups: sham-operated group,model group and acupuncture group.Only the left CCA,ECA and ICA were separated by sham surgery.MACO model was prepared in model group and acupuncture group.After the successful modeling of the rats in the acupuncture group,the rats were bound and pricked at 6h,24 h,48h and 72 h,respectively.The rats in the sham-operated group and the model group were bound but not pricked at the same time period.After the model was induced,the rats were assessed by the five-point scoring method and the stroke index scoring method at 2h,24 h,48h and 72 h after waking.TTC staining was used to examine the changes of cerebral infarction,HE staining was used to observe the changes of cerebral cortex,ELISA was used to detect The changes of Fas and fasl in rat serum,TUNEL was used to detect apoptosis,and immunofluorescence was used to detect caspase-3 and caspase-9 positive cells.Rusults: 1.Effect of acupuncture on neurological function score in MACO rats At 2h,24 h,48h and 72 h after the model was established,the neurological function of the rats was assessed by the five point scoring method and the stroke index scoring method.At 2h and 24 h,there was no significant difference between acupuncture group and model group(P > 0.05);at 48 h and 72 h,the analysis of difference was meaningful(P < 0.05).It can be concluded that acupuncture can significantly improve the neurological score of MACO rats 48 hours after intervention.2.Effects of acupuncture on the change of cerebral infarction in MACO rat After TTC staining,it was found that all the brain tissues in the sham-operated group were red,and no white infarcts were seen.After brain tissue staining in both the model group and the acupuncture group,the right hemisphere of the brain was red,while the left hemisphere of the brain was white,and the infarcts in the acupuncture group were significantly smaller than those in the model group.3.Effects of acupuncture on brain histomorphological changes in MACO rats Gross morphology of brain was observed by naked eye.In the sham-operated group,the left and right hemispheres were symmetrical,light red and evenly distributed.In the model group,the left and right hemispheres of brain were asymmetric,the color of infarcted brain tissue was pale,and a large area of brain tissue was swollen.In the acupuncture group,the left and right hemispheres were symmetrical,the infarcted side was pale,and the swelling of brain tissue was lighter than that in the model group.The changes of cerebral cortex in each group were observed after HE staining.In the Sham-operated group,nerve cells in the brain tissue were arranged in a dense and orderly manner.In the model group,a large number of nerve cells in the cerebral tissue of the infarcted side were denatured and necrotic,atrophic,with enlarged peripheral space,eosinophilic staining in cytoplasm,vacuolar denatured,loose,and hyperchromatic nuclei.In acupuncture group,the nerve cells in the infarct side of the brain were rank in a dense and orderly manner with a small surrounding space.4.Effects of acupuncture on serum Fas/Fasl levels in MACO rats Compared with the sham-operated group,the level of Fas and Fas L in the serum of the model group and the acupuncture rent rats increased(p<0.05);compared with the model group,the level of Fas and Fas L in the acupuncture group decreased significantly(p<0.05).5.Effect of acupuncture on the number of TUNEL positive cells of cerebralcortex in MACO rats There was a small amount of apoptotic cells in the sham-operated group,but more apoptotic cells in the model group and acupuncture group,and the analysis of difference was meaningful(P<0.05).However,compared with the model group,the expression of apoptotic cells in the acupuncture group decreased significantly(P<0.05).6.Effect of acupuncture on the expression of caspase-9,caspase-3 of cerebralcortex in MACO rats Compared with the sham-operated group,the expression of caspase-9 and caspase-3 positive cells in the model group and acupuncture group increased significantly,and the analysis of difference was meaningful(P < 0.05).However,compared with the model group,the expression of caspase-9 and caspase-3 positive cells in the acupuncture group decreased significantly(P < 0.05).Conclusion: 1.Acupuncture can effectively improve neurological function deficit in MACO rats.2.Acupuncture can reduce the cerebral infarction volume and the degeneration and necrosis of brain nerve cells in MACO rats.3.After acupuncture treatment,the content of Fas and Fasl in the serum of rats decreased,and the expression of caspase-3 and caspase-9 positive cells in the brain tissue of rats decreased,It was suggested that the intrinsic and extrinsic apoptotic pathways play an important role in the brain injury after cerebral ischemia and reperfusion in rats. |
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