Eye Acupuncture Regulates The Expression Of VEGF, Notch1, NICD And Hes1 In Brain Tissue Of Rats With Cerebral Ischemia-reperfusion Injury And Improves Microvessel Density

Purpose: Objective To observe the effect of eye acupuncture on the expression of VEGF,Notch1,Ni Cd,Hes1 and microvessel density(MVD)in ischemic penumbra of rats with cerebral ischemia-reperfusion injury(Ciri),and to explore the mechanism of eye acupuncture in improving cerebral ischemia-reperfusion injury.Method:Sixty rats were randomly divided into 10 normal group,10 sham operation group and 40 model group.In the sham operation group,the neck muscles were injured and sutured by blunt separation.The model rats were made by improved thread bolt method.After the model was successfully prepared,the neurological deficit was scored according to the longa5-point scoring system.After the defect score was completed,they were divided into model group,eye acupuncture group and external acupuncture group according to random number table.The rats in the eye acupuncture group and the external acupuncture group were treated with eye acupuncture three times at the moment of cerebral ischemia-reperfusion,24 hours and 30 minutes before sampling;the acupuncture points were liver and gallbladder area,upper Jiao area,lower Jiao area,kidney and bladder area.In the external acupuncture group,3mm outside the area of the same name in the eye acupuncture group was selected as the acupuncture site,and the acupuncture method was the same as that in the eye acupuncture area.The rats were killed with excessive hydrochloraldehyde.The neurological deficit score was performed again before death.MVD was detected by immunofluorescence staining,and the protein expression levels of VEGF,Notch1,Ni Cd and Hes1 were detected by Western blot.Results:1.The effect of eye on the neurological deficit score of Ciri rats;Compared with the sham operation group,the nerve defect scores of the model group,eye acupuncture group and external acupuncture group were significantly increased(P < 0.01),and the difference was statistically significant.Compared with the model group,the nerve defect score of the eye acupuncture group was significantly lower(P < 0.01),the difference was statistically significant;there was no significant difference in the nerve defect score of the external acupuncture group(P > 0.05),the difference was not statistically significant.Compared with the eye acupuncture group,the nerve defect score of the external acupuncture group was significantly higher(P < 0.01)2.The effect of eye acupuncture on MVD in ischemic penumbra of Ciri ratsCompared with the sham operation group,the number of CD31 positive cells in the ischemic penumbra of the model group and the external acupuncture group decreased significantly(P < 0.01),the difference was statistically significant;the number of CD31 positive cells in the ischemic penumbra of the eye acupuncture group increased significantly(P < 0.01),the difference was statistically significant.Compared with the model group,the number of CD31 positive cells in ischemic penumbra of eye acupuncture group was significantly increased(P < 0.01),the difference was statistically significant;the number of CD31 positive cells in ischemic penumbra of external acupuncture group had no significant change(P > 0.05),the difference was not statistically significant.Compared with the eye acupuncture group,the number of CD31 positive cells in the ischemic penumbra area of the external acupuncture group was significantly reduced(P < 0.01)3.The effect of eye acupuncture on VEGF protein expression in ischemic penumbra of Ciri ratsCompared with the sham operation group,the expression of VEGF in the ischemic penumbra of the model group and the external acupuncture group was significantly decreased(P < 0.01),and the difference was statistically significant;there was no significant difference in the expression of VEGF in the ischemic penumbra of the eye acupuncture group(P > 0.05),and the difference was not statistically significant;compared with the model group,the expression of VEGF in the ischemic penumbra of the eye acupuncture group was significantly increased(P < 0.01)There was no significant difference in the expression of VEGF in the ischemic penumbra of the external acupuncture group(P > 0.05),and the difference was not statistically significant;compared with the eye acupuncture group,the expression of VEGF in the ischemic penumbra of the external acupuncture group was significantly down regulated(P< 0.01),and the difference was statistically significant.4.Effect of eye acupuncture on Notch1 protein expression in ischemic penumbra of Ciri ratsCompared with the sham operation group,the expression of Notch1 in the ischemic penumbra of the model group,eye acupuncture group and external acupuncture group was significantly decreased(P < 0.01),and the difference was statistically significant;compared with the model group,the expression of Notch1 in the ischemic penumbra of the eye acupuncture group was significantly increased(P < 0.01),and the difference was statistically significant,There was no significant difference in the expression of Notch1 in the ischemic penumbra of the external acupuncture group(P > 0.05),and the difference was not statistically significant;compared with the eye acupuncture group,the expression of Notch1 in the ischemic penumbra of the external acupuncture group was significantly down regulated(P < 0.01),and the difference was statistically significant.5.Effect of eye acupuncture on the expression of Ni Cd protein in ischemic penumbra of Ciri ratsCompared with the sham operation group,the expression of Ni Cd in the ischemic penumbra of the model group,eye acupuncture group and external acupuncture group was significantly decreased(P < 0.01),and the difference was statistically significant;compared with the model group,the expression of Ni Cd in the ischemic penumbra of the eye acupuncture group was significantly increased(P < 0.01),and the difference was statistically significant,There was no significant difference in the expression of Ni Cd in the ischemic penumbra of the external acupuncture group(P > 0.05),and the difference was not statistically significant;compared with the eye acupuncture group,the expression of Ni Cd in the ischemic penumbra of the external acupuncture group was significantly down regulated(P< 0.01),and the difference was statistically significant.6.Effect of eye acupuncture on Hes1 protein expression in ischemic penumbra of Ciri ratsCompared with the sham operation group,the expression of Hes1 in the ischemic penumbra of the model group,eye acupuncture group and external acupuncture group was significantly decreased(P < 0.01),and the difference was statistically significant;compared with the model group,the expression of Hes1 in the ischemic penumbra of the eye acupuncture group was significantly increased(P < 0.01),and the difference was statistically significant,Compared with the eye acupuncture group,the expression of Hes1 in the ischemic penumbra of the external acupuncture group was significantly decreased(P < 0.01),and the difference was statistically significant.Conclusion:1.Eye acupuncture can increase microvessel density in ischemic penumbra of Ciri rats2.Eye acupuncture can improve microvessel density by up regulating the expression of VEGF,Notch1,Ni Cd and Hes1 in ischemic penumbra of Ciri rats.