| Objective:To observe the effects of acupuncture-rehabilitation therapy on motor function and pathological damage of brain tissue of rats with focal cerabral ischemia,and to clarify the effects of acupuncture-rehabilitation therapy on lipid peroxidation and the mechnism of cell resistance to ferroptosis by detecting iron metabolism and lipid metabolism related proteins,so as to provide a theoretical basis for the prevention and treatment of cerebral ischemia.Methods:180 male rats were were randomly divided into 5 groups,namely sham group,model group,acupuncture group,rehabilitation group,and acupuncture-rehabilitation group,and each group was randomly divided into three subgroups on the basis of 3d,7d and 14 d time points(n=12).In later four groups,the blood flow of the middle cerebral artery was blocked by the modified line plug method,and a permanent rat cerebral ischemia model(MCAO)was established.In the sham group,the line plug was inserted but did not block the blood flow of the middle cerebral artery.The acupuncture group was treatede with scalp acupuncture traeatment,the rehabilitation group with running platform training,and the acupuncture-rehabilitation group used scalp acupuncture combined with running platform training,the sham group and model group were only performed under the same conditions without any treatment.At 3d,7d and14 d after surgery,modified neurological deficit score(m NSS)and HE staining were used to evaluate brain injury,and the iron deposition in the ischemic brain tissue of rats was detected by tissue iron kit.The expression of ROS,MDA,GPX4 and SLC7A11 in the ischemic area of rats were detected in ELISA,and the protein expression of P53,SLC7A11,ACSL4,and COX2 were tested by WB.Results:1.Modified neurological severity scores: At 3d,7d and 14 d after surgery,m NSS score of the rats in the model group increased significantly compared with the sham group(P<0.05);Compared with the model group,the m NSS score decreased in each treatment group(P<0.05),and the m NSS score in the acupuncture-rehabilitation group was lower than the single acupuncture and rehabilitation group(P<0.05).2.Pathological changes of ischemic brain tissue in rats: At 3d,7d and 14 d after surgery,compared with the sham group,rats in the model group showed obvious infarction,and the pathological damage of the ischemic side cortex was significantly worse.Compared with the model group,the degree of brain tissue pathological damage in each treatment group was reduced,among which the effect was the most obvious in the acupuncture-rehabilitation group,and the volume of cerebral infarction was decreased.3.Iron deposition in ischemic brain tissue cells: At 3d,7d and14 d after surgery,the iron content in the model group increased sig nificantly compared with the sham group(P<0.05);Compared with t he model group,the iron content in the ischemic brain tissue decre ased in each treatment group(P<0.05),with the most significant de crease in the acupuncture-rehabilitation group(P<0.05).4.Expressions of ROS and MDA in ischemic brain tissue: At 3d,7d and 14 d after surgery,the levels of ROS and MDA in the model group was significantly increased compared with the sham group(P<0.05);The amount of ROS and MDA in brain tissue of each treatment group was lower compared with the model group(P<0.05),and the amount of ROS and MDA in ischemic brain tissue of acupuncture-rehabilitation group was lower than that of acupuncture and rehabilitation group(P<0.05).5.Expressions of GPX4 and SLC7A11 in ischemic brain tissue: At3 d,7d and 14 d after surgery,the amount of GPX4 and SLC7A11 in the model group was significantly reduced compared with the sham group(P<0.05);The amount of GPX4 and SLC7A11 in each treatment group increased compared with the model group(P<0.05),and the content of GPX 4 and SLC7A11 in the acupuncture-rehabilitation group was higher than the acupuncture and rehabilitation group(P<0.05).6.Expression of P53 and SLC7A11 protein in ischemic brain tissue:At 3d,7d and 14 d after surgery,compared with the sham group,the expression of P53 in the ischemic brain tissue was increased in the model group,and the expression of SLC7A11 in the ischemic brain tissue was decreased in the model group(P<0.05);Compared with the model group,the P53 decreased in the brain and the SLC7A11 increased in each treatment group(P<0.05);Compared with the treatment groups,the expression of P53 decreased and the SLC7A11 increased significantly in the ischemic area(P<0.05).7.Expression of ACSL4 and COX2 protein in ischemic brain tissue:At 3d,7d and 14 d after surgery,the expression of ACSL4 and COX2 increased in the ischemic area of the model group compared with the sham group(P<0.05);the content of ACSL4 and COX2 in each treatment group decreased compared with the model group(P<0.05),and the expression of ACSL4 and COX2 in the acupuncture-rehabilitation group was significantly lower compared with the acupuncture group and rehabilitation group(P<0.05).Conclusions:1.Acupuncture-rehabilitation therapy can reduce the degree of pathological damage of ischemic side cortex and reduce the infarct area,so as to improve the neurological dysfunction of cerebral ischemic rats.2.Acupuncture-rehabilitation therapy can upregulate the expression of antioxidant enzymes GPX4 and SLC7A11 protein,and reduce the content of ROS and MDA in the ischemic brain tissue,thus reducing the lipid peroxidation damage of cerebral ischemia;3.Acupuncture-rehabilitation therapy can inhibit the expression of P53,ACSL4 and COX2 protein in ischemic brain tissue,and reduce intracellular iron deposition,thus inhibiting ferroptosis and improving cerebral ischemic injury. |
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