| The first part Eye acupuncture combined with exercise therapy on MCAO model rat pia mater and ophthalmic artery collateral circulation current intervention effect of experimental researchPurpose: Explore the eye acupuncture combined with exercise therapy on MCAO model the intervention of the collateral circulation in rats, the eye acupuncture combined with exercise therapy on the cerebral cortex and ophthalmic artery flow intervention mechanism.Materials and methods: Using the random number table method will randomly drew only 20, 170 rats were divided into blank group and 10 were only and control, the remaining 150 reproducing line plug method MACO rats model, copy the model group rats by using line switch on cerebral ischemia reperfusion model replication. Model to copy from rat survival after scoring standard 138 rats, were randomly divided into 4 groups: model control group only 35, head needle group and 35 group and 34, eye, eye needle movement group of 34 only. Reperfusion after 1 h, after being wide-awake rats for neural function defect scale. Reperfusion group begin to eye needle, needle eye exercise therapy group, the first needle acupuncture treatment group accordingly. Reperfusion 3 h, 24 h, 72 h, respectively to the corresponding point in time, the use of laser speckle imaging technology monitoring pia mater and ocular blood flow velocity of the conjunctival surface, after pulpotomy executed immediately stripped whole brain tissue, TTC staining.Results: 1.The model of evaluation result, replication compared with blank group, modelgroup rats behavior score was statistically significant(p < 0.01);Replication compared with control group, model group rats behavior score was statistically significant(p < 0.01). Whole brain TTC Dyeing, according to the results of model replication group of rats appeared obvious infarcts, and the blank group and control group did not see infarcts occur. 2.The results showed that the blank group, control group each time point observation regional blood flow velocity is almost no change; Compared with the blank group, model group, needle head, eye needle, needle eye movement group at all time points observed regional blood flow velocity were increased significantly(p < 0.05); Compared with model group, eye needle all time points observed regional blood flow velocity were increased significantly(p < 0.05) needle eye movement group all time points observed regional blood flow velocity were increased significantly(p < 0.01); Needle eye movement group than other groups of blood flow increase significantly(p < 0.01). 3.The results show that the blank group and control group of conjunctiva blood flow velocity at various time points have no obvious change. Compared with the blank group, model group, needle head, eye needle, needle eye movement group at all time points for conjunctiva blood flow velocity changes have significantly increased(p < 0.05, p < 0.01); Compared with model group, eye needle, needle eye movement group at all time points for conjunctiva blood flow velocity changes have significantly increased(p < 0.01); Needle eye movement group at all time points for conjunctiva blood flow velocity changes significantly higher than other groups(p < 0.05, p < 0.01).Conclusion: 1.This experiment through the nerve function score and TTC staining method validation MCAO rats model building. Using laser speckle imaging technology validation and eye acupuncture combined with exercise therapy on MCAO rats model eye conjunctiva blood flow and blood flow, the influence of respectively on MCAO model in the rat brain cortex, whole blood eye conjunctiva blood flow were monitored.2.The experimental results show that the eye acupuncture combined with exercise therapy can improve model of MCAO rats cortex blood flow velocity percentage of 3 h began to increase, to 72 h showed a trend of increased gradually. And eye acupuncture combined with exercise e exercise therapy can improve the percentage of eye conjunctiva blood flow velocity, began to gain 3 h, 24 h, the height of the 72 h decline(but improve the 3 h). Prompt and Eye acupuncture combined with exercise therapy may by increasing the cerebral cortex blood flow velocity and increase the ophthalmic artery flow to open the secondary collateral circulation, thus improving cerebral ischemia reperfusion regional ischemia and hypoxia.The second part Eye acupuncture combined with exercise therapy on paper 2 ischemia half dark area of MCAO rats brain tissue VEGF/VEGFR signal transduction pathwayPurpose: Exploring the eye acupuncture combined with exercise therapy ischemia half dark area of MACO model rats brain tissue VEGF/VEGFR signal transduction pathway.Materials and methods: Using the random number table method will randomly drew only 20, 170 rats were divided into blank group and 10 were only and control, the remaining 150 reproducing line plug method MACO rats model, copy the model group rats by using line switch on cerebral ischemia reperfusion model replication. Model to copy from rat survival after scoring standard 138 rats, were randomly divided into 4 groups: model control group only 35, head needle group and 35 group and 34, eye, eye needle movement group of 34 only. 1 hours after ischemia reperfusion, reperfusion group begin to eye needle, scalp needle group, eye acupuncture combined with exercise therapy group on corresponding acupuncture and exercise therapy. Groups of rats in 3 h reperfusion, 24 h, 72h, 10% chloral hydrate after intraperitoneal injection of anesthesia, abdominal aorta puncture whole blood collected 5 ml, using enzyme-linked immunosorbent(ELISA) method to detect each group HIF- 1 content in serum of rats; Ischemia pulpotomy executed after half dark band organization, the Western- bloting method and immunohistochemical method and ischemia rats half dark with VEGF and VEGFR2 in regional organizations, Notch protein expression level; Detection by using RT-PCR method, groups of ischemia rats half dark with regional organizations in VEGF m RNA and VEGFR2 m RNA, Notch m RNA expression level.Results: 1.The results showed that compared with the blank group, model control group, the head of the needle, needle eye group, the eye acupuncture combined with exercise group HIF- 1 content in serum of rats increased obviously(p < 0.01); Compared with model group, group, eye eye needle therapy combined with exercise group content increased obviously, was statistically significant(p < 0.01); Compared with eye acupuncture combined with exercise group and the blank group, control group, model group, head of the needle, needle eye group HIF- 1 content have obvious differences, statistically significant(p < 0.01). 2.The results show that compared with the blank group, model group, the group, the eye head needle group, eye acupuncture combined with exercise group of ischemia rats half dark zone.Area in the brain tissue significantly increases the expression level of VEGF and VEGFR2, Notch1(p < 0.01); Compared with model group,eye needle group and eye movement group had a significantly increase the expression level of VEGF and VEGFR2, Notch1(p < 0.05). 3.The results show that, compared with the blank group, model group, the group, the eye head needle group, eye acupuncture combined with exercise group of ischemia rats half dark zone.Area in the brain tissue VEGF and VEGFR2, Notch1 m RNA expression level rise significantly(p < 0.01); With the model group than A group and eye acupuncture therapy combined with exercise group of VEGF and VEGFR2, Notch1 m RNA expression level rise significantly(p < 0.05).Conclusion: 1.The blank group, the control group there was no significant alteration of the expression of HIF- 1, the model group with HIF- 1 expression, the expression of ischemia and hypoxia can stimulate. Group, needle eye, eye head needle therapy combined with exercise group of cerebral ischemia reperfusion 3 h increased expression of HIF- 1, peaked at 24 h, 72 h express slightly down(is still higher than 3 h). Group, needle head, eye eye needle therapy combined with exercise group and model group HIF-l expression trend is the same, but the needle eye movement group of HIF- 1 at each time point than a significant rise in expression, MACO shows that eye acupuncture combined with exercise treatment group rats after can promote the expression of HIF- 1, thus significantly improve brain neurons to hypoxia and ischemia tolerance, play the role of brain protection. 2.The cerebral ischemia reperfusion rats in 3 h to detect the expression of VEGF and VEGFR2, Notch1, confirmed that cerebral ischemia reperfusion injury induced the expression of VEGF and VEGFR2, Notch1. Eye acupuncture combined with exercise therapy in 3 h, 24 h, 72 h point can significantly increase VEGF and VEGFR2, Notch1 and its m RNA expression, peaked at 72 h, proof needle eye exercise therapy through the VEGF/VEGFR expression Notch1 and upstream factor to promote angiogenesis, increase the blood perfusion and oxygen supply, promote the opening of the collateral circulation, improve brain function. 3.Eye acupuncture combined with exercise therapy by VEGF/VEGFR signal conduction system promote the proliferation of endothelial cells and small blood vessels of the newborn, save the ischemic tissue surrounding the neuronal survival and regeneration, which promotes neural functional recovery.The third part Eye acupuncture combined with exercise therapy ischemia half dark area of MCAO model rats brain tissue Ang/Tie-2 the influence of signal transduction pathwaysPurpose: Exploring the rat MCAO model of ischemia half dark area of brain tissue Ang/Tie- 2 the activation of signal transduction pathways, the eye acupuncture combined with exercise therapy effect on the spread system. Materials and methods: Using the random number table method will randomly drew only 20, 170 rats were divided into blank group and 10 were only and control, the remaining 150 reproducing line plug method MACO rats model, copy the model group rats by using line switch on cerebral ischemia reperfusion model replication. Model to copy from rat survival after scoring standard 138 rats, were randomly divided into 4 groups: model control group only 35, head needle group and 35 group and 34, eye, eye needle therapy combined with exercise group 34 only. Groups of rats in 3 h reperfusion, 24 h, 72 h, 10% chloral hydrate after intraperitoneal injection of anesthesia, abdominal aorta puncture whole blood collected 5 ml, using enzyme-linked immunosorbent(ELISA) method to detect each group rats serum Ang-1, Ang-2 levels; Ischemia pulpotomy executed after half dark band organization, the Western- bloting method and immunohistochemical method and ischemia rats half dark area in the organization, Ang- 2 and Tie2 Ang- 1 protein expression level; Detection by using rt-pcr method, groups of ischemia rats half dark with regional organizations in m RNA, Ang Ang- 1-2 m RNA, Tie2 m RNA expression level.Results: 1.The blank group, the control group at all time points Ang no change-1 content; Model group, the group, the eye head needle group, the eye acupuncture combined with exercise group all time points increased gradually, Ang-1 h 72 peak; Compared with the blank group, model group, the group, the eye head needle group, eye acupuncture combined with exercise group content increased significantly, Ang-1 was statistically significant(p < 0.01); Compared with model group, group, eye eye needle therapy combined with exercise group Ang-1 content significantly increased, with statistical significance(p < 0.01); Compared with needle eye movement group and other groups Ang-1 content of each time point compared withsignificant difference(p < 0.01). 2.The results show that compared with the blank group, model group, the group, the eye head needle group, the eye acupuncture combined with exercise group rats ischemic half dark area in the brain tissue, Tie2 Ang-1 expression level rise significantly(p < 0.01); Compared with model group, group and eye eye needle therapy combined with exercise group, Ang-1/Tie2 expression level rise significantly(p < 0.05). 3.The results show that, compared with the blank group, model group, the group, the eye head needle group, the eye acupuncture combined with exercise group rats ischemic half dark area in the brain tissue, Ang-1/Tie2 m RNA expression level rise significantly(p < 0.01); Compared with model group, group and eye eye needle therapy combined with exercise group, Ang- 1 Tie2 m RNA expression level rise significantly(p < 0.05).Conclusion: 1.The rat MCAO model in 3 h time Ang- 1 and its m RNA expression level rise, with the extension of time Ang- 1 and its m RNA expression gradually increased, 72 h peak value, and through the combined with Tie- 2 receptor, promote angiogenesis, prompt cerebral ischemia half dark factors associated with angiogenesis, start the angiogenesis and reshape the ischemic penumbra area. 2.The eye acupuncture combined with exercise therapy can promote to some extent Ang- 1 content in serum, increasing Ang- 1 and its m RNA expression, combine with Tie- 2 receptor, start Ang/Tie- 2 signal transduction system, regulation of ischemia on time and the number of half dark with the formation of new blood vessels and collateral circulation, improve the cerebral blood flow of ischemic penumbra area, thus inhibiting neuronal apoptosis, restore the cerebral blood flow, promote the recovery of neural function after cerebral ischemia.
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