Effects Of Yiqi Huoxue Prescription On Autophagy And LC3 Protein Expression In Rats With Cerebral Ischemia-reperfusio

Objective: 1.Observe the activation of autophagy in different phases after successful ischemia reperfusion rat molding,and explore the relationship between autophagy and cerebral ischemic injury.2.To investigate the effect of Yiqi Huoxue Recipe on the regulation of autophagy and LC3 protein expression in ischemic reperfusion rats.Methods: 90 male SD rats of SPF grade were selected,based on the principle of random control,of which 30 were enrolled in the sham group,and the remaining 60 were established with a rat MCAO model by wire bolt method,and after 2 hours,the line plug was pulled out to form ischemia and then perfused.After the model was successfully prepared,it was then divided into 30 Model group and YHR group.Each group was randomly divided into three phases according to the time of execution: 3d,7d,and 14 d.Sham group,Model group gastric irrigation and other volumes of distilled water,YHR group irrigation of equal volume of Yiqi Huoxue Recipe.The extent of neurologic deficit in rats was assessed with m-NSS scores;the expression of autophagy-tagged protein LC3 was detected by hematoxylin-eosin(HE)staining;the number of autophagosomes(AP)was observed by transmission electron microscopy(TEM);and the expression of autophagy-labeled protein LC3 was detected by western blot(WB).The statistical results of the experimental data are expressed in mean ±standard deviation(x±s),and the one-way ANOVA method is selected for statistics,and the Bonfrenny method is applied when the two-by-two comparison between the groups and the variance are uniform,and the Tamheini method is applied when the variance is uneven.p<0.05 suggests that the difference is statistically significant.Results: 1.m-NSS scores: The score of the Sham group is 0,and there are no obvious symptoms of neurological defects.Both the Model group and the YHR group showed obvious neurological defect symptoms(p<0.05,n=6),indicating successful mold making.Compared with the Model group,the scores of the YHR group decreased(p<0.05,n=6),suggesting that YHR can promote the recovery of neural function in MCAO rat defects.2.HE:There is no obvious damage to the neurons of the cerebral tissue in the cortical layer of the Sham group,there is no abnormality in cell morphology and structure,the arrangement is regular,the appearance of the cell body is fusiform or round,there is a large nucleus,and the nerve fiber lines are basically the same.Compared with the Sham group,after the successful preparation of the model,the morphology of brain tissue cells in the rat cortex showed obvious pathological changes: the cell morphology was very different,the structure was disordered,the arrangement was irregular,the number of neurons was reduced,the cell envelope was incomplete,the cell body was triangular,the pericellular space was expanded,the nuclear solidification was dark,and the nerve fibers were chaotic.Compared with the Model group,there were fewer pathological changes in the morphology of brain tissue cells in each phase of the YHR group.3.AP: A small number of AP formations can be seen in the Sham group;Compared with the Sham group,the number of apps in the three phases of the Model group increased(P<0.05,n=6);the number of APs in Model 3d,Model 7d,and Model 14 d was not significantly different(P>0.05,n=6).The above results show that autophagy in brain tissue after focal cerebral ischemia is activated,but there is no obvious dynamic change law in 3d,7d and 14 d.Compared with the Model group,the number of AP in each phase of the YHR group decreased(p<0.05,n=6),suggesting that YHR has an inhibitory effect on autophagy.4.WB: LC3 protein in the Sham group had a certain expression;compared with the Sham group,the expression of LC3 protein in each phase of the Model group was upregulated(P<0.05,n=6);compared with the Model group,the LC3 expression of YHR group in each phase was down-regulated(P<0.05,n=6).It shows that YHR can downregulate LC3 protein.Conclusion: After the successful preparation of the ischemic reperfusion rat model,autophagy was activated,and there was no obvious dynamic change law in 3d,7d,and 14 d,and the degree of autophagy activation was positively correlated with the degree of neural function defect in rat focal areas.Yiqi Huoxue Recipe can inhibit autophagy by downregulating the LC3 protein,and improve the symptoms of ischemia and reperfusion of local nerve defects in rats.