| Purpose:This study applies eye acupuncture combined rehabilitation therapy to treat patients with motor dysfunction during the recovery period of ischemic stroke.The therapeutic effect is evaluated by observing the patient’s motor function,daily living ability,and serological indicators.By observing the intervention of eye acupuncture combined rehabilitation therapy on MCAO/R rats,observe the changes of nerve function,motor function and pathological morphology of brain tissue of rats in each group,explore its protective mechanism on brain function of MCAO/R rats,and explore the effect of eye acupuncture combined rehabilitation therapy on VEGFR2/PLCγ/PKC pathway from the mechanism of angiogenesis.Material and method:1.122 patients were hospitalized from December 2020 to December 2021 at the Rehabilitation Center of Liaoning University of Traditional Chinese Medicine Affiliated Hospital,Brain Disease Rehabilitation Department II,and the Rehabilitation Department of Shengjing Hospital Affiliated to China Medical University.They were diagnosed as patients with motor dysfunction during the recovery period of ischemic stroke.They were randomly divided into experimental group and control group,with 61 cases in each group.During the experiment,there were 2 cases of detachment in the experimental group and 1 case of detachment in the control group,with 119 cases actually completed.There is no significant difference in gender,age,or course of disease between the two groups,and the general data is comparable between groups.The experimental group received eye acupuncture combined rehabilitation therapy,while the control group received routine rehabilitation therapy.Both groups were treated with basic internal medicine and necessary body acupuncture according to the guidelines.The frequency of rehabilitation treatment for both groups is once a day,with5 treatments per week and a course of 28 days.We used the Simplified Fugl Meyer Motor Function Scale(FMA),Modified Barthel Index Rating Scale(MBI),Traditional Chinese Medicine Syndrome Score,and serum BDNF and VEGF levels as evaluation indicators.2.We randomly divided the rats into blank group,sham group,model group,eye acupuncture combined rehabilitation(ER)group,and ER+SU5416 group,each consisting of 20 rats.The establishment of a cerebral ischemia-reperfusion model was carried out using an improved thread occlusion method.Intervention measures:Blank group:without any treatment,raised normally.Sham group:Following the model surgery method,the insertion depth of the suture was controlled to prevent the formation of cerebral ischemic lesions.Model group:After the MCAO/R model is successfully replicated,no processing is done and it is raised normally.ER group:After successful replication of the MCAO/R model,eye acupuncture combined rehabilitation therapy was performed after cerebral ischemia-reperfusion,once every 12 hours,twice a day,for a total of 7 days.ER+SU5416 group:Inject VEGFR2 blocker(SU5416)into the abdominal cavity 30 minutes before rehabilitation treatment with eye acupuncture combined rehabilitation,and the rest are the same as the ER group.Observation indicators:a,Neurological function evaluation:We used the Zea Longa Neurological Function Deficit Score to score each group of rats,ensuring the success of model establishment and comparability of baseline values among each group.The m NSS neurological deficit score was used to evaluate the neurological function of rats in each group after reperfusion and 7 days of treatment.b,The TTC staining method was used to observe the infarct area of rats in each group after 7 days of treatment,and the recovery of brain tissue in each group was observed by calculating and comparing the infarct area.c,A rat grip tester was used to measure the grip of each group of rats before modeling,after reperfusion,and after 7 days of treatment,to observe the recovery of motor function in each group of rats.d,Immunofluorescence method was used to measure the expression of HIF-1αprotein in the ischemic penumbra of rats in each group.Real time q PCR reaction was used to detect the relative expression of VEGF and VEGFR2 m RNA in the ischemic penumbra.Western blotting was used to measure the relative expression of VEGF,VEGFR2,PLCγ1,p-PLCγ1,PKC,p-PKC in the ischemic penumbra of rats in each group.Immunohistochemical method was used to measure the positive cells of CD34~+.Results:1.General data:There was no significant difference in age,gender,and course of disease between the two groups of patients(P>0.05),and there was comparability between the groups.2.There was no statistically significant difference in the upper and lower limb FMA motor function scores between the two groups of patients before treatment(P>0.05);Compared with before treatment,there was a significant difference in upper and lower limb motor function scores between the control group and the experimental group after treatment(P<0.01);Compared with the control group,there was a significant difference in the motor function scores of the upper and lower limbs in the experimental group after treatment(P<0.01).3.There was no statistically significant difference in the activity of daily living(ADL)scores between the two groups of patients before treatment(P>0.05);Compared with before treatment,there was a significant difference in ADL scores between the control group and the experimental group after treatment(P<0.01);Compared with the control group,there was a significant difference in ADL scores after treatment in the experimental group(P<0.05).4.There was no statistically significant difference in the TCM syndrome scores between the two groups of patients before treatment(P>0.05);Compared with before treatment,there was a significant difference in the TCM syndrome scores between the control group and the experimental group after treatment(P<0.01);Compared with the control group,there was a significant difference in the score of traditional Chinese medicine syndromes after treatment in the experimental group(P<0.05).5.After treatment,3 patients in the control group showed significant clinical efficacy and 44patients were effective,with a total effective rate of 81.03%;After treatment,6 patients in the experimental group showed significant clinical efficacy and 48 patients were effective,with a total effective rate of 91.53%.6.There were significant differences in serum BDNF and VEGF levels between the control group and the experimental group after treatment(P<0.01);Compared with the control group,there was a significant difference in serum BDNF levels after treatment in the experimental group(P<0.01),while there was no statistically significant difference in serum VEGF levels(P>0.05).7.The Zelongga score of rats in the Sham group remained unchanged,while the Zelongga score of rats in the Model group,ER group,and ER+SU5416 group significantly increased after modeling(P<0.01),indicating the success of cerebral ischemia-reperfusion modeling in rats.8.The 7-day survival rate of ER group rats was higher than that of Model group and ER+SU5416 group,and the survival rate decreased slowly compared to Model group and ER+SU5416 group.9.After 7 days of intervention,compared with the Blank group,there was no significant change in TTC staining volume in the Sham group rats after modeling(P>0.05);Compared with the Sham group,the TTC staining volume percentage of the Model group rats significantly increased after modeling(P<0.01);Compared with the Model group,the TTC staining volume percentage of ER group rats significantly decreased(P<0.01);Compared with the ER group,the ER+SU5416 group showed a significant increase in TTC staining volume percentage(P<0.01).10.After reperfusion,compared with the Sham group,the model group,ER group,and ER+SU5416 group significantly reduced the forelimb grip strength of rats(P<0.01).There was no statistically significant difference(P>0.05)among the model group,ER group,and ER+SU5416 group.After 7 days of modeling,compared with the Sham group,the Model group,ER group,and ER+SU5416 group significantly reduced the forelimb grip strength of rats(P<0.01);Compared with the Model group,the ER group showed a significant increase in forelimb grip strength(P<0.01);Compared with the ER group,the ER+SU5416 group showed a decreasing trend in forelimb grip strength,but the difference was not statistically significant(P>0.05).11.Compared with the Blank group,there was no significant change in the m NSS scores of the Sham group rats after cerebral ischemia-reperfusion and 7 days after modeling(P>0.05);Compared with the Sham group,the m NSS score of the Model group rats significantly increased at 7 days after reperfusion(P<0.01);Compared with the Model group,the m NSS score of ER group rats showed a downward trend after 7 days,but the difference was not statistically significant(P>0.05).In inter group comparison,compared with reperfusion,the m NSS score in the ER group significantly decreased after 7 days of modeling(P<0.01),while the m NSS score in the ER+SU5416 group showed a downward trend after 7 days of modeling,but the difference was not statistically significant(P>0.05).12.Immunohistochemical results showed that there were fewer CD34+cells in the ischemic penumbra tissue of the Blank and Sham groups of rats;Compared with the Blank group,the Model group showed an increase in the number of CD34+cells in the ischemic penumbra tissue;Compared with the Model group,the ER group showed an increase in the number of CD34+cells in the ischemic penumbra tissue;Compared with the ER group,the number of CD34+cells in the ischemic penumbra tissue of the ER+SU5416 group decreased.13.There was no significant change in the relative expression levels of VEGF and VEGFR2m RNA in the ischemic penumbra tissue of rats in the Blank and Sham groups(P>0.05);Compared with the Sham group,the relative expression level of VEGFR2 m RNA in the Model group was significantly increased(P<0.01),while the relative expression levels of VEGF and VEGFR2 m RNA in the ER group were significantly increased(P<0.01);Compared with the Model group,the relative expression levels of VEGF and VEGFR2m RNA in the ER group increased(P<0.05,P<0.01);Compared with the ER group,the relative expression levels of VEGF and VEGFR2 m RNA in the ER+SU5416 group decreased(P<0.05,P<0.01).14.Immunofluorescence results showed that the protein expression of HIF-1αin the ischemic penumbra tissue of rats in the Blank and Sham groups was lower;Compared with the Blank group,the protein expression of HIF-1αin the ischemic penumbra tissue of the Model group increased;Compared with the Model group,the protein expression of HIF-1αin the ischemic penumbra tissue of the ER group increased;Compared with the ER group,the protein expression of HIF-1αin the ischemic penumbra tissue of the ER+SU5416 group decreased.15.The relative expression levels of VEGF,VEGFR2,p-PLCγ1,PLCγ1,p-PKC,and PKC proteins in the ischemic penumbra tissue of rats in the Blank and Sham groups showed no significant changes(P>0.05);Compared with the Sham group,the relative expression levels of VEGF and VEGFR2 proteins in the Model group increased(P<0.05,P<0.01),while the relative expression levels of VEGF,VEGFR2,p-PLCγ1/PLCγ1,p-PKC/PKC proteins in the ER group significantly increased(P<0.01);Compared with the Model group,the relative expression levels of VEGF,VEGFR2,and p-PLCγ1/PLCγ1 proteins in the ER group increased(P<0.05,P<0.01),while the relative expression levels of p-PKC/PKC proteins showed an upward trend,but the difference was not statistically significant(P>0.05);Compared with the ER group,the relative expression levels of VEGF,VEGFR2,p-PLCγ1/PLCγ1,and p-PKC/PKC proteins in the ER+SU5416 group decreased(P<0.05).Conclusion:1.Through clinical research,it has been confirmed that eye acupuncture combined rehabilitation therapy can promote the recovery of motor function disorders in patients with ischemic stroke during the recovery period,significantly enhance motor function,improve clinical symptoms and signs of traditional Chinese medicine,and improve daily living ability.2.Eye acupuncture combined rehabilitation therapy can improve the levels of serum BDNF and VEGF factors in patients with ischemic stroke,indicating that eye acupuncture with needle rehabilitation therapy may play a role in neuroprotection and promoting angiogenesis.3.Animal experiments have shown that eye acupuncture combined rehabilitation therapy can improve the cerebral infarction area of MCAO/R rats,promote motor and neurological function recovery.4.Eye acupuncture combined rehabilitation therapy promotes angiogenesis by activating the VEGFR2/PLCγ/PKC signal pathway,which may be one of the mechanisms of eye acupuncture combined rehabilitation therapy promoting the recovery of motor function in ischemic stroke. |
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