| Objective:To observe the degree of damage to the nervous system of cerebral ischemia-reperfusion rats by electroacupuncture,by detecting the metabolism and oxidative stress levels of iron and other related proteins,the protective effect of electroacupuncture on cerebral ischemia/reperfusion injury and its effects on iron metabolism and oxidative stress levels were clarified,provides evidence for the prevention and treatment of ischemic stroke.Methods:The 240 male rats were grouped by random number table method,according to this method,the rats were divided into 5 groups.There are sham operation group,model group,electroacupuncture pretreatment(p-EA)group,electroacupuncture treatment(t-EA)group,and all terms electroacupuncture(a-EA)group,24 animals in each group.The p-EA group and the a-EA group were pretreated with electroacupuncture for 30min every day for 2 weeks.The rat middle cerebral artery ischemia model was established 24 hours after the last electroacupuncture preconditioning in each group,and 12 hours after cerebral ischemia-reperfusion,the t-EA group and a-EA group were given electroacupuncture treatment for 30 minutes per day.Three days after reperfusion,the infarct volume on the ischemic side of the rat brain was measured by TTC staining,and the degree of pathological damage of the tissue on the ischemic side of the brain was observed by HE staining.The expression of FPN-1 protein in the ischemic lateral brain tissue was detected by Western Blot,and the content of Hepcidin protein in the infarcted tissue was detected by ELISA.Tissue iron kit was used to detect the content of iron ion in the brain tissue of the infarcted lesion.The degree of neurological damage in rats will be assessed by the modified neurological deficit score after 1d,3d,and 7d of reperfusion.The expressions of ROS,GSH and GPX4 in the brain tissue of the infarcted area were detected by ELISA.To observe the effect of electroacupuncture on cerebral infarct size,FPN-1 protein expression,Hepcidin content,iron content and ROS,GSH,GPX4 content in acute cerebral ischemia-reperfusion rats in order to clarify that electroacupuncture can regulate the iron metabolism reduces oxidative stress injury caused by iron overload after cerebral ischemia-reperfusion and exerts a protective effect on the brain.Results:1.TTC staining showed that after 3d of cerebral ischemia and reperfusion,the rats in the other groups except the sham group had obvious infarcts.Among the EA groups,the area of the a-EA group was the smallest,followed by the p-EA group,and the largest in the t-EA group(P<0.05).2.HE staining showed that after 1d and 3d of cerebral ischemiareperfusion,a large number of pathological damages occurred in the model group.The damage in the p-EA group and t-EA group was smaller than that in the model group,and the damage in the a-EA group was the smallest.3.The results of iron content in brain tissue showed that after 1d and 3d of cerebral ischemia-reperfusion,the iron content in model group increased significantly(P<0.05);the content in p-EA group was lower than that in model group(P<0.05);t-EA group was lower than model group(P<0.05)but had no statistical difference compared with p-EA group;a-EA group was lower than model group and t-EA group at 1d(P<0.05),3d time was lower than that of other EA groups(P<0.05).4.The results of the content of Hepcidin in brain tissue showed that after 1d and 3d of cerebral ischemia-reperfusion,the content of Hepcidin in the brain tissue of the model group increased significantly(P<0.05);the content of the p-EA group was lower than that of the model group(P<0.05);t-EA group was lower than model group(P<0.05)but had no statistical difference compared with p-EA group;a-EA group was lower than model group and t-EA group at 1d(P<0.05),3d time was lower than that of other EA groups(P<0.05).5.The expression of FPN-1 in brain tissue showed that the expression of FPN-1 in the brain tissue of the model group was significantly decreased after 1d and 3d of cerebral ischemia-reperfusion(P<0.05);the expression of p-EA group was higher than that of the model group(P<0.05);the t-EA group was higher than the model group(P<0.05)but had no statistical difference compared with the p-EA group;the a-EA group was higher than the model group and t-EA group at 1d(P<0.05),higher than other EA groups at 3d(P<0.05).6.The results of mNSS score showed that there was no increase in the scores of the rats in the sham group,but the scores of the other groups increased to a certain extent(P<0.05).Except that there was no difference between the t-EA group and the model group on the 1d,the scores of all EA groups at each time point were lower than those of the model group(P<0.05).Both group and t-EA group had lower scores(P<0.05).7.The results of GSH and GPX4 content in brain tissue showed that after 1d,3d,7d of cerebral ischemia-reperfusion,the content of GSH and GPX4 in brain tissue of model group decreased significantly(P<0.05);the content of p-EA group was higher than that of model group(P<0.05);there was no statistical difference between the t-EA group and the model group at 1d,and it was higher than the model group at 3d(P<0.05),but there was no statistical difference compared with the p-EA group,and at 7d It was higher than the model group and p-EA group(P<0.05);the a-EA group was higher than the model group and t-EA group(P<0.05)at 1d,but there was no statistical difference compared with the p-EA group,and at 3d it was higher than other electroacupuncture groups(P<0.05),and the content of a-EA group was not significantly different from that of the sham group at 7days(P<0.05).8.The results of ROS content in brain tissue showed that after 1d,3d,and 7d of cerebral ischemia-reperfusion,the expression of ROS in model group increased significantly(P<0.05);the content of p-EA group was lower than that of model group(P<0.05);the t-EA group had no statistical difference compared with the model group at 1d,was lower than the model group at 3d(P<0.05)but had no statistical difference compared with the pEA group,and was lower than the model group at 7d and p-EA group(P<0.05);a-EA group was lower than model group and t-EA group(P<0.05)at 1d,but had no statistical difference compared with p-EA group,and was lower at 3d and 7d in other electroacupuncture groups(P<0.05).Conclusions:1.Electroacupuncture and electroacupuncture preconditioning can reduce acute cerebral ischemia-reperfusion injury and have neuroprotective effects;2.Electroacupuncture and electroacupuncture preconditioning can reduce iron overload by regulating iron metabolism after cerebral ischemiareperfusion;3.Electroacupuncture and electroacupuncture preconditioning can exert neuroprotective effects by reducing oxidative stress injury caused by iron overload after cerebral ischemia-reperfusion;4.Both electroacupuncture and electroacupuncture preconditioning can protect nerves and reduce cerebral ischemia-reperfusion injury,and the combination of the two can exert a more significant effect. |
PDF Full Text Request