| Purpose: To evaluate the efficacy of rehabilitation with eye-acupuncture on motor dysfunction of ischemic stroke,and to explore its anti-apoptotic and protective mechanisms for neurons in ischemic areas through the PI3K/Akt signal transduction pathway,and restore the nerve function of ischemic stroke.Material and method:A total of 90 patients with motor dysfunction after ischemic stroke who were admitted to the Second Department of Encephalopathy rehabilitation,Rehabilitation Center,Affiliated Hospital of Liaoning University of Traditional Chinese Medicine from January 2021 to December 2021 were randomly divided into eye acupuncture group,eye acupuncture group and control group,with 30 patients in each group.During the study,2 cases of shedding in the rehabilitation with eye-acupuncture group(leaving the hospital for personal reasons),3 cases of shedding in the eye acupuncture group(including 2 cases of aggravation,including 1 case of self-withdrawal),3 cases of shedding in the control group(including 2 cases of leaving the hospital for personal reasons,including 1 case of aggravation),actually completed 82 cases.There was no significant difference in gender,age,stroke-meridian syndrome classification(P>0.05),and the comparability between groups was good.Rehabilitation with eye-acupuncture group was given rehabilitation with eye-acupuncture combined with conventional rehabilitation,eye acupuncture combined with conventional rehabilitation,and control group with conventional rehabilitation.Conventional rehabilitation treatment includes: 1.basic drug therapy;2.routine rehabilitation training:exercise therapy,action therapy,electric starting bed,low-frequency pulse electrical therapy;3.conventional body acupuncture treatment.The frequency of the above treatment was once a day and the course of treatment was 4 weeks,5 times a week and 2 times of rest.One rehabilitation assessment before treatment,14 days after treatment,and 28 days after treatment,using the National Institutes of Health Stroke Scale(NIHSS Scale),simplified Fugl-Meyer Motor Function Scale,Modified Barthel Index Rating Scale(MBI),Stroke TCM syndrome score is used as an evaluation index.SPSS 23.0 software was used to perform statistical analysis.Animal experiments using random number method from 70 SD rats randomly selected 16 sham surgical groups,4 normal groups,50 model replication groups,the model replication group rats using the improved wire plug method for cerebral ischemia reperfusion model replication,successful 46,each group randomly selected 1 for TTC staining,and then the model replication group was randomly divided into: 15 model groups,15 rehabilitation with eye-acupuncture groups,and 15 blocker groups.Interventions: Normal group: no treatment,normal feeding.Sham surgical groups: no treatment is done after surgery,normal feeding.Model group: After the MCAO model is successfully copied,no treatment is done and it is normally reared.Rehabilitation with Eye-acupuncture group: After successful replication of the MCAO model,rehabilitation with eye-acupuncture was performed 1h after cerebral ischemia and reperfusion,every 12 h twice a day for a total of 3 days.Blocker group:After successful replication of the MCAO model,the rehabilitation with eye-acupuncture was injected with PI3 K blocker(LY294002)through the enterocoelia 20 minutes before treatment,and rehabilitation with eye-acupuncture was performed 1h after cerebral ischemia and reperfusion,every 12 h twice a day for a total of 3 days.Observation indicators: 1.Neural function evaluation: The neural function evaluation of rats in the model replication group(rehabilitation with eye-acupuncture group,blocker group,model group)was evaluated at 1h and 72 h of ischemia and reperfusion using neural defect score.2.Tunel method to detect apoptosis: after ischemia reperfusion for 72 h,5 rats from each group(sham surgical group,rehabilitation with eye-acupuncture group,blocker group,model group)were randomly selected for apoptosis cell detection.3.At 72 h of ischemia reperfusion,10 rats were selected from each group(sham surgical group,rehabilitation with eye-acupuncture group,blocker group,model group),and the expression of PI3 K,p-Akt,caspase3,Bcl-2,Bax in ischemic semi-dark band region of the hippocampus was detected by Western Blot method.Real-time PCR reaction(RT-PCR)was used to detect the expression of m RNA in the ischemic semi-dark band region of the hippocampus Bcl-2,Bax,caspase3.Results:1.General data: There were no significant differences in age,sex and stroke-meridian syndrome types among the three groups(P > 0.05).2.NIHSS score: Before treatment,there was no significant difference in NIHSS scores among the three groups(P>0.05).In the comparison of the group,after 14 days and 28 days of treatment the NIHSS scores of all three groups decreased(P<0.05).Compared with each group,after 14 and 28 days of treatment,NIHSS score in the eye acupuncture group was significantly lower than that in the control group(P < 0.05),and NIHSS score in the rehabilitation with eye-acupuncture group was significantly lower than that in the eye acupuncture group(P < 0.05).3.FMA score: Before treatment,there was no significant difference in FMA scores among the three groups(P>0.05).In the comparison of the group,after 14 days and 28 days of treatment,the FMA scores of the three groups were improved(P<0.05).Compared with each group,after 14 days and 28 days of treatment,the FMA score in the eye acupuncture group was significantly higher than that in the control group(P < 0.05),and FMA score in the rehabilitation with eye-acupuncture group was significantly higher than that in the eye acupuncture group(P < 0.05).4.MBI score: Before treatment,there was no statistically significant difference in FMA scores among the three groups(P>0.05).In the comparison of the group,after 14 days and 28 days of treatment,the MBI scores of the three groups were improved(P<0.05).Compared with each group,after 14 days and 28 days of treatment,the MBI score in the eye acupuncture group was significantly higher than that in the control group(P < 0.05),and MBI score in the rehabilitation with eye-acupuncture group was significantly higher than that in the eye acupuncture group(P < 0.05).5.Stroke TCM syndrome score: Before treatment,there was no statistically significant difference in stroke TCM syndrome scores among the three groups(P>0.05).Compared with the group,after 14 days and 28 days of treatment,stroke TCM symptom scores decreased in the three groups(P<0.05).Compared with each group,after 14 days and 28 days of treatment,stroke TCM symptom scores in eye acupuncture group was significantly decreased compared with the control group(P < 0.05),and the stroke TCM symptom scores in the rehabilitation with eye-acupuncture group was significantly decreased compared with the eye acupuncture group(P < 0.05).After 28 days of treatment,the effective rate of eye acupuncture group was89.29%(P < 0.05).6.TTC staining results of whole brain tissue showed obvious infarcts in model replication group,but no infarct in normal group and sham surgical group.7.After 1h of ischemia reperfusion,there was no significant difference in the neural defect score of the three groups(rehabilitation with eye-acupuncture,blocker group,model group)which was comparable(P > 0.05).After 72 h of ischemia reperfusion,neural defect score of the eye acupuncture group was significantly lower than that of the blocker group and the model group,the difference was statistically significant.(P < 0.05)8.Five visual fields were taken in the CA1 area of the right hippocampus of each rat.The TUNEL method showed that compared with the model group and the blocker group,the apoptotic cells in the rehabilitation with eye-acupuncture group were significantly reduced(P< 0.05),but there was no significant difference between the model group and the blocker group(P > 0.05).9.After ischemia reperfusion for 72 h,compared with the sham surgical group,the expression levels of p-PI3 K and p-Akt proteins in the model group decreased significantly,and the difference was statistically significant(P<0.05);compared with the model control group,rehabilitation with eye-acupuncture was significantly increased by p-PI3 K.p-Akt protein expression(P<0.05);intraperitoneal injection of LY294002 abolished the role of rehabilitation with eye-acupuncture,indicating that LY294002 inhibited PI3 K phosphorylation,and thus inhibited the role of p-PI3 K in rehabilitation with eye-acupuncture anti-apoptotic and alleviating deficiency and reperfusion damage.Illustration:the rehabilitation with eye-acupuncture is expressed by activating the PI3K/Akt signal transduction pathway and increasing p-PI3 K and p-Akt.There were no significant effects on total PI3 K and Akt proteins in the sham surgical group,rehabilitation with eye-acupuncture group,blocker group and model group(P>0.05).10.After ischemia reperfusion for 72 h,compared with the sham surgical group,the expression level of Bcl-2 protein in the model group decreased significantly,and the expression of Bax and caspase3 proteins increased significantly,and the difference was statistically significant(P<0.05);compared with the model group,the rehabilitation with eye-acupuncture group significantly increased the expression of Bcl-2 protein(P<0.05),which reduced Bax,Caspse3 protein expression(P<0.05);intraperitoneal injection of LY294002 blocker abolished the enhancing effect of rehabilitation with eye-acupuncture on Bcl-2 protein,indicating that LY294002 inhibited PI3 K phosphorylation and then inhibited Bcl-2 protein expression,which increased the expression of Bax protein and promoted the upregulation of caspase3 protein expression.Illustration: rehabilitation with Eye-acupuncture inhibits the expression of Bax and caspse3 proteins by activating the PI3K/Akt signal transduction pathway.There were no significant differences in the effects of blocker group and model group on Bcl-2,Bax,caspase3 proteins(P>0.05).11.After ischemia reperfusion for 72 h,rehabilitation with eye-acupuncture increased the m RNA level of Bcl-2 and decreased the m RNA level of Bax and caspase3 compared with the model group,and the difference was statistically significant(P<0.05).Compared with the sham surgical group,the m RNA levels of Bcl-2 were reduced in the model group and the blocker group,and the m RNA levels of Bax and caspase3(P<0.05)were increased,and the difference was not statistically significant between the model group and the blocker group(P>0.05).Compared with the rehabilitation with eye-acupuncture group,the blocker group injected LY294002 blocker intraperitoneally,which reduced the m RNA level of Bcl-2 and increased the m RNA level of Bax and caspase3,indicating that the rehabilitation with eye-acupuncture promotes the transcription of Bcl-2 by activating the PI3K/Akt signal transduction pathway.Inhibits transcription of Bax and caspase3.Conclusion:1.Through randomized controlled clinical research,rehabilitation with eye-acupuncture can promote the recovery of motor function of ischemic stroke,significantly reduce nerve defect degree,the clinical symptoms and signs of TCM were improved,daily life ability enhancement,in the early ischemic stroke rehabilitation plays an important role.2.Animal experiments show that with rehabilitation with eye-acupuncture can significantly reduce the number of apoptosis of hippocampus ischemia area,promote cell survival,restore nerve function injury in rats,rehabilitation with eye-acupuncture through antiapoptotic role play to the effect of brain protection,improve the ability of neurons in ischemic tolerance,promote neurological function recovery of ischemic stroke.3.Rehabilitation with eye-acupuncture by activating PI3K/Akt signal transduction pathway,increase anti-apoptotic protein expression,reduce promoting apoptosis protein expression,inhibition of the mitochondrial mediated apoptosis pathway,blocking apoptotic cascade,alleviate the nerve injury caused by ischemia,achieve nerve protection,is the mechanism of neurological function recovery in ischemic stroke. |
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