| Purpose:Based on the research of ancient literature,this paper discusses the correlation between kidney deficiency and blood stasis,kidney brain dysfunction and ischemic stroke,and the advantages of eye acupuncture treatment;Neurological deficit score,open field test,hematoxylin eosin staining(HE),Nissl staining and 5-bromo-2-fluorouracil staining were detected 5-bromo-2'-deoxy uridine(Brd U)labeled neurons and other indicators were used to observe the neuroprotective effect of eye acupuncture intervention on cerebral ischemia-reperfusion injury(CI/RI)rats;By detecting the expression levels of A2B5,O4,CNPase,MBP,Olig2,MAG and other proteins in brain tissue,the effect of eye acupuncture intervention on the maturation and activation of oligodendrocytes was expounded;By detecting Wnt,DVL and GSK-3???-catenin to explore the mechanism of eye acupuncture in promoting oligodendrocyte maturation and activation and accelerating myelin regeneration by regulating Wnt signaling pathway,so as to further explore the role of eye acupuncture in"kidney brain interaction"through kidney tonifying and blood activating therapy,so as to provide new ideas for clinical application of eye acupuncture characteristic therapy of traditional Chinese medicine.Material and method:1.Read the database of mental diseases of traditional Chinese medicine of Liaoning University of traditional Chinese medicine,analyze the relevant ancient books,summarize the ancient doctors'understanding of eye acupuncture,five wheel and eight outline theory and stroke,and explore the feasibility of eye acupuncture therapy based on the theory of"five wheel and eight outline"and combined with the theory of"kidney and brain"to treat stroke of kidney deficiency and blood stasis type.2.In this study,60 adult SPF SD rats,half male and half female,were selected as the research objects.60 rats were marked as No.1-60 according to their body weight,and then they were randomly divided into two groups,namely sham operation group(n=12);Model group(48rats).Methods:48 rats in the model group were used to establish CI/RI model by suture method.The model rats were evaluated by neurological function score combined with TTC(triphenyl tetrazolium chloride)staining and he staining.The rats with successful model evaluation were randomly divided into three groups(random number method),namely model control group;Inhibitor group;Eye acupuncture group.Interventions:in the eye acupuncture group,the rats were treated with eye acupuncture at 2 h,12 h and 24 h after cerebral ischemia-reperfusion,and the acupuncture points were selected from the kidney area,upper Jiao area,liver area and lower Jiao area;The rats in the inhibitor group were given Wnt-c59orally 24 hours before sampling,and the rats in the sham operation group and the model control group were fed normally.The rats in the inhibitor group were detected by behavioral test(neurobehavioral score,open field test),morphological test(HE staining,Nissl staining,Brd U labeling),and oligodendrocyte activation related proteins(A2B5,O4,CNPase,MBP,Olig2 The key factors of Wnt signaling pathway(Wnt,DVL,GSK-3???-catenin)The aim of this study was to observe the effect of eye acupuncture on oligodendrocytes in CI/RI rats,and to explore the mechanism of acupuncture therapy on brain protection of CI/RI rats.Results:1.Literature analysis:through combing the ancient literature,we believe that kidney deficiency and blood stasis,kidney and brain dysfunction are the main pathogenesis of ischemic stroke,and Peng's eye plays its advantages and role in the treatment of this disease.2.CI/RI model rats was successfully reproduced by modified suture method:the neurological function score of model group was significantly higher than that of sham operation group(P<0.01).TTC staining results of brain tissue showed that there was no obvious abnormality in the sham operation group,and there was a white hypoxic-ischemic area in the right brain tissue of the model group.He staining results of brain tissue showed that the structure of nerve cells in the right hippocampus of the sham operation group was intact,while the right brain tissue of the model group was damaged in varying degrees.3.Eye acupuncture therapy improves cognitive impairment of CI/RI model rats:compared with sham operation group,the neurological function scores of model control group,inhibitor group and eye acupuncture group were significantly increased 2 hours after cerebral ischemia-reperfusion(P<0.01);After eye acupuncture treatment,the neurological function score of the eye acupuncture group was significantly decreased(P<0.01).The results of open field experiment showed that before eye acupuncture intervention,compared with the sham operation group,the dwell time and movement distance of the model control group,inhibitor group and eye acupuncture group were significantly reduced(P<0.01).After eye acupuncture intervention,compared with the sham operation group,the dwell time and movement distance of middle grid in model control group,inhibitor group and eye acupuncture group were significantly reduced(P<0.01);Compared with the model control group,the dwell time and movement distance of the eye acupuncture group were significantly increased(P<0.01).He staining results of brain tissue showed that after eye acupuncture treatment,compared with the control group,the damage degree of brain tissue in the inhibitor group was significantly increased,the water content of brain tissue in the eye acupuncture group was decreased,and various pathological changes were improved in varying degrees.4.Eye acupuncture therapy promotes the regeneration of neurons in CI/RI model rats:Nissl staining results showed that the structure of neurons in the sham operation group was complete,there were more positive cells,and the Nissl bodies of neurons were clear and orderly arranged;Compared with the sham operation group,the arrangement of neurons in the model control group was sparse,and some Nissl bodies were fuzzy;Compared with the model control group,the pathological changes of nerve cells in the eye acupuncture group were significantly reduced,and the number of nerve cells was increased,with statistical significance(P<0.01).Brd U labeling test results showed that compared with sham operation group,the number of Brd U positive cells in brain tissue of model control group,inhibitor group and eye acupuncture group was significantly reduced(P<0.01);Compared with the model control group,the number of Brd U positive cells in the inhibitor group was significantly decreased(P<0.01),and the number of Brd U positive cells in the eye acupuncture group was significantly increased(P<0.01).5.The expression of activated oligodendrocyte related proteins in eye acupuncture group:the results of immunofluorescence detection of A2B5,O4 and CNPase showed that compared with the sham operation group,the expression levels of activated oligodendrocyte related proteins A2B5,O4 and CNPase in model control group,inhibitor group and eye acupuncture group were significantly down regulated(P<0.01);Compared with the model control group,the expression levels of A2B5,O4 and CNPase in the inhibitor group were significantly decreased(P<0.01),and the expression levels of A2B5,O4 and CNPase in the eye acupuncture group were significantly increased(P<0.01).The results of MBP,Olig2 and MAG showed that compared with the sham operation group,the expression levels of MBP,Olig2 and MAG in the model control group,inhibitor group and eye acupuncture group were significantly down regulated(P<0.01);Compared with the model control group,the expression levels of MBP,Olig2 and MAG in the inhibitor group were significantly decreased(P<0.01),and the expression levels of MBP,Olig2 and MAG in the eye acupuncture group were significantly increased(P<0.01).6.Eye acupuncture activates Wnt signaling pathway to promote neuronal regeneration in CI/RI rats:the key proteins of Wnt pathway were detected by immunohistochemistry and Western blotting.The results showed that compared with sham operation group,Wnt,Dvl,and Wnt in brain tissue of model control group,eye acupuncture group and inhibitor group were significantly increased?-catenin was significantly decreased(P<0.01)GSK-3?was significantly increased(P<0.01);Compared with the model control group,Wnt,Dvl,?-catenin was significantly higher than that of GSK-3?(P<0.01)The expression level of Wnt,Dvl,?-catenin in brain tissue of the inhibitor group was significantly lower than that of the control group(P<0.01)?-catenin was significantly decreased(P<0.01)GSK-3?The protein expression level was significantly increased(P<0.01).Conclusion:1.This study combs the ancient literature,and finds that kidney deficiency and blood stasis,kidney and brain dysfunction are the main pathogenesis of ischemic stroke.Peng's eye plays its advantages in the treatment of this disease.2.By observing the effect of eye acupuncture intervention on CI/RI rats,it is confirmed that eye acupuncture therapy can promote the repair of brain tissue after CI/RI,restore neural function,and improve cognitive dysfunction after ischemic stroke.3.Eye acupuncture can promote the expression of Nissl bodies,OLs and their related proteins,accelerate the formation of myelin sheath,promote the regeneration of neurons and play a neuroprotective role in CI/RI rats.4.Eye acupuncture can promote the regeneration of nerve myelin sheath by activating Wnt signaling pathway,so as to play a neuroprotective role. |
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