| Objective:A rat model of cerebral ischemia and reperfusion was prepared by the suture method,with the inflammatory response-mediated apoptosis of nerve cells as the starting point for the research,and the PARP-1 signaling pathway as the research focus,to explore the combination of Tibetan medicine in the treatment of bulgur disease The protective effect and mechanism of the regimen(in the morning,morning and evening,respectively,Qishiwei Pearl Pills,Ruyi Zhenbao Pills,and Twenty Wei Chenxiang Pills)on cerebral ischemia-reperfusion injury in rats and its mechanism,which provides a valuable resource for the treatment of cerebral ischemia.Medical program.Methods:The animal experiment was carried out for the Tibetan medicine combination regimen.The middle artery embolism reperfusion rat cerebral ischemia model was prepared by the thread embolization method.After the model was successfully established,the neurological deficit was scored according to the Longa score method,and the scores 1 to 3 were included in the subsequent study.The rats were randomly divided into blank group,sham operation group,model group,positive control group,Qishiwei pearl pill group,two prescription combination medication group and three prescription combination medication group.Each group was divided into 4 subgroups according to the reperfusion time.The groups are 1d,3d,7d and 15d.The detection methods of this experiment are as follows:①General inspection:Record the activity state,fur state,spirit,diet and weight of rats in each group.② Nerve function test:The rats in each group were scored for neurological deficit and balance beam walking test on 1d,3d,7d,15d after operation.③Brain tissue pathological examination:TTC staining method was used to determine the cerebral infarction rate of rats in each group,and HE staining was used to observe the cerebral ischemic core,tissue edema,neuronal pyknosis and softening of the cortex under an optical microscope.④Detection of cell apoptosis:TUNEL staining method was used to detect the dynamic changes of cell apoptosis in the ischemic penumbra of the right cerebral cortex of rats.⑤ PARP-1 signaling pathway:using immunohistochemistry to detect the dynamic changes of PARP-1,NF-κB,TNF-α,IL-6,P53,Bax,Fas in the penumbra of cerebral ischemia in the right cerebral cortex of rats.Results:①General inspection:The activity status,fur status,mental diet status,and weight status of rats in each group showed differences with the increase of time.The normal coat color of the rats in the three-way combination group was similar to that of the sham operation,and the mental and diet status was similar to that of the sham operation.The body weight basically returned to normal at 15 days.The rats in the other groups showed varying degrees of yellowish coat,poor mental diet,and weight loss.Among them,the model group was the worst.According to the results of weight statistical analysis:the weight of the rats in the three-way combination medication group was higher than that of the model group at each time point,and all were significant(P<0.05);the average weight of the rats in the three-way combination medication group was higher than positive at 3d,7d,and 15d There was a statistically significant difference between the control group,the Qishiwei Pearl Pill group and the two prescription combination medication group(P<0.05).The three prescription combination medication group had a better weight recovery effect.②Neural function test:The neurological deficit score shows that the scores of the three-way combination medication group are lower than the model group at each time point,and there is a significant difference at 15d(P<0.05);the scores of the three-way combination medication group on 7d and 15d are lower than the positive control The scores of the 15-day tripartite combination medication group were lower than the Qishiwei Zhenzhu pill group and the second prescription combination medication group,but there was no statistical difference(P>0.05).The behavioral scores showed that the behavioral scores of rats in the tripartite combination medication group were lower than the model group and the positive control group at each time point,and there was a significant difference between 7d and 15d(P<0.05).At each time point,the behavior of rats in the three-prescription combination group was lower than that of the Qishiwei Zhenzhu pill group and the two-prescription combination group,and there was a significant difference at 15 days(P<0.05);③Brain tissue pathology examination:TTC staining found that each At the time point,the cerebral infarction area of the rats in the three prescription combination group was smaller than that of the model group,and there were significant differences(P<0.05).The results of HE staining showed that the degree of ischemic cavities,cell rupture,and softening of the rats in the three-prescription combination group were lower than those in the model group and the positive control group;at 7 and 15 days of reperfusion,the three-prescription combination group had better efficacy than rats Seventy flavor pearl pill group and two prescription combination medication group.④Apoptosis detection:The number of TUNEL-positive cells in the dark ischemic zone of the right cerebral cortex of rats in the three-way combination medication group at each time point was lower than that in the model group(P<0.05).Compared with the positive control group,the number of TUNEL positive cells in the three-party combination group was reduced,and there were significant differences on 3d,7d,and 15d(P<0.05).Compared with the Qishiwei pearl pill group and the second prescription combination drug group,the number of TUNEL positive cells in the three prescription combination drug group was lower,and there was a significant difference between 7d and 15d(P<0.05).⑤PARP-1 signaling pathway:The expressions of PARP-1,NF-κB,TNF-α,IL-6,P53,Bax and Fas in the dark ischemic zone of the right cerebral cortex of rats in the three-way combination treatment group at each time point were significantly lower In the model group(P<0.05);compared with the positive control group,PARP-1,NF-κB,TNF-α,IL-6,P53,Bax in the dark ischemic zone of the right cerebral cortex of rats in the three-way combination treatment group The expression of Fas and Fas protein decreased to varying degrees.Bax protein was significantly decreased at four time points(P<0.05),PARP-1 and IL-6 protein at 7d were significantly decreased(P<0.05),and NF-at 3d and 15d κB protein was significantly reduced(P<0.05),TNF-α protein at 15d was significantly reduced(P<0.05),and P53 and Fas protein at 3d,7d,and 15d were significantly reduced(P<0.05).Conclusion:The combination of Tibetan medicine has a protective effect on ischemia-reperfusion injury in rats.The three-way combination regimen has the best effect on the recovery of neurological function in rats with cerebral ischemia-reperfusion.The main mechanism may be:this therapy is effective in cerebral ischemia reperfusion in rats.After blood reperfusion,by regulating the PARP-1 signaling pathway,inhibit the expression of PARP-1 and its downstream proteins,inhibit the inflammatory cascade mediated by NF-κB,TNF-α,and IL-6 inflammatory cytokines,thereby inhibiting P53,Bax,Fas protein-mediated apoptosis,and ultimately reduce inflammatory factor-mediated apoptosis of nerve cells,reduce neuronal damage after cerebral ischemia,thereby reducing the area of cerebral infarction in rats after cerebral ischemia,and improving rats Limb function has a protective effect on cerebral ischemia-reperfusion injury in rats. |
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