| Objective: To study the effect of Qingnao Yiyuan Decoction on the expression of neuropeptides NPY and CGRP after focal cerebral ischemic injury in rats,and to explore the mechanism of Qingnao Yiyuan Decoction in preventing and treating focal cerebral ischemic injury.Methods: 240 SD rats were stratified according to body weight,and then divided into two groups,A and B,120 rats in each group,and each group was divided into 4 groups using random number table method: blank group group,sham operationgroup,modelgroup,Qingnao Yiyuan Tanggroup.Except for the blank group,the rats in each group made the middle cerebral artery embolism model according to the Longa intraluminal thread embolization method.Among them,the sham operation group inserted a wire plug 8-10 mm into the internal carotid artery of the rat.This depth is not enough to block the blood flow of the middle cerebral artery.The remaining steps are the same as the operation group.According to Longa’s 5-point system,the rats were scored for neurological deficits and divided into 5subgroups according to the five sacrifice time points of 1d,3d,7d,14 d,and28d after MCAO operation,with 6 rats in each subgroup.The blank groupwas fed normally and eat freely.Rats in the sham operation group and model group were given an equal volume of distilled water after surgery,and the Qingnao Yiyuan decoction group was given an equal volume of Qingnao Yiyuan decoction,which lasted until each time point of taking materials.HE staining method was used to detect the morphological changes of rat nerve cells;immunohistochemistry method was used to detect the expression level of NPY immunopositive cells in ischemic brain tissue sections of each group of rats;Western blot method was used to detect CGRP Protein content,and SPSS24.0 statistical software package for data processing.Results:(1)Neurological deficit score:(1)There was no neurological deficit in rats in the sham operation group at all time points.(2)Compared with the sham operation group,the neurological deficit scores of the model group and Qingnao Yiyuan group were significantly increased,and the differences were statistically significant(P <0.01).(3)Compared with the model group,the neurological deficit symptoms of rats in Qingnao Yiyuan group were improved to different degrees at the same time point,and the difference was statistically significant(P <0.01).(2)HE staining:(1)Blank group,sham operation group: no edema of brain tissue,cells are neatly arranged,and their morphological structure is basically normal.(2)Model group: Rat neurons shrunken and deformed,arranged loosely in triangles,scattered widely in the necrotic red neurons,most neurons were degenerated and necrotic,the gaps around the cells increased,the cell body deformed and shrunk,and the nucleus was solidly contracted Infiltration and necrosis of inflammatory cells,cell dissolution disappears or a small amount of solid nucleus remains.(3)Qingnao Yiyuan Decoction group: After administration of the drug,a small amount of neuronal necrosis,nuclear solid shrinkage and other pathological features can be seen in the infarcted area of the brain tissue,which is significantly improved compared with the model group,and the neuron cells in the cortical area are closely arranged and form a spherical shape,Cell arrangement tends to be normal,and occasionally some cells are deformed.It shows that Qingnao Yiyuan Decoction can promote the repair of neuron cells,or it can regenerate nerve cells and repair nerve damage.(3)Immunohistochemical method to detect NPY:(1)A small amount of NPY immunopositive cells were observed in the blank group and sham-operated rat brain tissue at each time point,and there was no statistically significant difference between the two groups at the same time(P> 0.05).(2)At the same time point,compared with the sham operation group,the number of NPY positive cells in the model group and Qingnao Yiyuan decoction group increased significantly,and the difference was statistically significant(P <0.01).(3)At the same time point,compared with the model group,the number of NPY immunopositive cells in the Qingnao Yiyuan Decoction group was significantly lower than that in the modelgroup,the difference was statistically significant(P <0.01).(4)Western blot detection of CGRP:(1)CGRP is expressed in a small amount in the brain tissue of the blank group and the sham operation group.There is no significant difference between the two(P> 0.05).(2)At the same time point,compared with the sham operation group,the CGRP protein expression of the model group and the Qingnao Yiyuan decoction group were increased(P <0.01),the difference was statistically significant.(3)Compared with the model group,the relative expression of CGRP protein in the brain tissue of the Qingnao Yiyuan decoction group was significantly higher than that of the model group,and the difference was statistically significant(P <0.01).Conclusion: Qingnao Yiyuan Decoction can improve the damage of cerebral ischemic tissue in rats,and its mechanism may be to reduce the number of NPY positive cells and increase the expression of CGRP protein after cerebral ischemic injury,thereby regulating the vasomotor state of cerebral blood vessels and improving the deficiency.Cerebral blood flow in the blood area. |
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