Mechanism Research Of Fire Needle On Improving Neural Function In Rats With Cerebral Infarction Based On The Regulation Of Autophagy

ObjectiveObserve the effect of fire needle on the behavior,the percentage of cerebral infarction volume,the morphology and structure of cerebral vascular endothelial cells,and the expression of key proteins in autophagy in rats with cerebral infarction at different time points.Explore the mechanism of fire needle in improving the brain nerve function after cerebral infarction.Methods150 SD rats were randomly divided into three groups: sham operation group(1d,3d,7d),model group(1d,3d,7d),fire needle group(1d,3d,7d).The rats in the fire needle group and the model group were used to prepare MCAO model by the modified autothrombotic method.In the fire needle group,he’s fine fire needle was used to acupuncture Baihui(GV20),Quchi(LI11),Hegu(LI4),Zusanli(ST36)and Taichong(LR3)points,while the sham operation group and model group were not given therapeutic intervention.At the corresponding time,the rats in each group were assessed with longa behavioral score.The whole brain was stained by TTC staining,then photographed,and the image was measured with Image-Pro Plus 6.0software,and then the infarct volume percentage was calculated.The micromorphology and ultrastructure of vascular endothelial cells in each group of rats were observed with optical microscope and transmission electron microscope.Western blot was used to detect the expression of autophagy-related proteins(PI3KC3、Beclin-1、LC3Ⅱ、AKT).Results(1)Behavioral score: The scores of rats in the sham operation group were all 0,indicating that there was no neurological impairment.Compared with it,the longa score of nerve function in the model group was significantly higher at three time points,the difference was statistically significant(P=0.000<0.05),and there were symptoms of nerve defect such as limb flexion and weakness on the opposite side,turning and dumping to the opposite side during walking,and the score gradually decreased at 3d and 7d over time,but the difference was not statistically significant.The Longa score of the fire needle group decreased significantly over time,and it was significantly lower than that of the model group at 7d,and the difference was statistically significant(P=0.015<0.05).(2)Cerebral infarction volume percentage: The brain tissue staining of the sham operation group showed no white infarcted area.In contrast,the infarcted area of the model group and the fire needle group was clearly visible.Over time,the percentage of infarct volume in the model group increased to some extent at 3d and 7d,but the difference was not statistically significant.The percentage of infarcted area in the fire needle group also increased slightly at 3d and 7d,and the infarct percentage at the three time points was smaller than that of the model group,but the difference was not statistically significant.(3)Micromorphology and ultrastructure of vascular endothelial cells: In the sham operated group,the cerebral vascular endothelial cells of the rats were tightly and orderly connected,completely and seamlessly lining the inner wall of the vessels.The cells are smooth and fusiform,with intact membrane,rich and translucent cytoplasm,complete organelle components,large nucleus,intact nuclear membrane,small proportion of nucleoplasm and obvious nucleolus.In the model group,the vascular endothelial cells fell off from the inner wall of the blood vessel,arranged disorderly and gathered in the lumen.The cells were swollen and abnormal in shape,the integrity of the cell membrane was destroyed,the organelles were seriously damaged and lost,and the nucleus was swollen.However,a certain degree of change occurred over time: the degree of aggregation was reduced,the degree of swelling of cells and nuclei was reduced,the integrity of the cell membrane was gradually restored,and the nuclei and organelles were restored to a certain extent.In the fire needle group,after the fire needle treatment,the morphology of vascular endothelial cells basically returned to fusiform,the integrity of cell membrane was restored,the damaged organelles and nuclei were improved,and gradually recovered.(4)Occurrence of autophagy: Transmission electron microscopy showed that autophagy of vascular endothelial cells occurred obviously in the fire needle group,and the number of autophagy increased significantly at 3d and 7d.After 1 day of middle cerebral artery occlusion,compared with the sham operation group,the PI3KC3,Beclin-1,and LC3Ⅱproteins in the model group were all increased,and the expression levels increased more significantly over time,and the difference was statistically significant(P<0.05).It is suggested that these proteins are activated after the middle cerebral artery is occluded,indicating that autophagy participates and runs through the dynamic process of acute cerebralinfarction.After fire needle therapy intervention for 1d to 3d,the autophagy-related proteins PI3KC3,Beclin-1,and LC3Ⅱ all increased significantly,and the trend continued to 7d,the difference was statistically significant(P<0.05).ConclusionFire needle therapy can promote the recovery of neural function in rats with cerebral infarction,and its mechanism may be related to up regulating the expression of autophagy related proteins PI3KC3 、 Beclin-1 and LC3 Ⅱ,regulating the autophagy of vascular endothelial cells,and improving the injury of vascular endothelial cells.