| Objective:To observe the effects of electroacupuncture on the learning and memory ability of middle cerebral artery occlusion(MCAO)model rats and on the expression of key genes and proteins in the CACNα1D-HSPB1-CAMSAP2-SNCG pathway in the hippocampus,and to explore the improvement of learning and memory by electroacupuncture The mechanism of ability provides a theoretical and experimental basis for the clinical application of electroacupuncture to treat vascular cognitive impairment(VCI).Methods:Thirty-six SD male rats with a body weight of(280±20)g were randomly divided into sham operation group,model group and electroacupuncture group,with 12 rats in each group.Model group and electroacupuncture group refer to Longa’s modified suture method for MCAO model preparation.The sham operation group was sutured af ter blunt separation of blood vessels,and the rest of the operations were the same as in the model group.The electro-acupuncture group uses electro-acupuncture at Shenti ng and Baihui points for intervention.Electro-acupuncture is applied with a density wave with a frequency of 2-10 Hz and a current intensity of 1.5-2 m A.The intensity is gradually increased to the point where the rat’s ears are slightly jittery and do not scream.,Stimulation time 20 min,24h after surgery,start treatment,once a day for 7days,the positive and negative poles of each electroacupuncture are different from the previous one.The model group and the sham operation group received the same conditions as the electro-acupuncture group and then returned to the cage without treatment.On the 1st and 7th day after operation,follow the Zea-Longa method to perform neurobehavioral score and balance beam test;perform water maze test on the 3-7d postoperative day;each group will be randomly selected after the end of the electroacup uncture intervention on the 7th postoperative day Three mice were stained with 2,3,5-Triphenyte trazoliumchloride(TTC)to observe cerebral ischemia;real-time fluorescence quantitative polymerase chain reaction(Real-time polymerase chain reaction)reaction,RT-PCR)to detect the m RNA expression of hippocampal CACNα1d,HSPB1,CAMSAP2,and SNCG;Western blot(WB)to detect the expression of CACNα1d,HSPB1,CAMSAP2,and SNCG in the hippocampus;immunohistochemical detection of CACNα1d,HSPB1,CAMSAP2,and SNCG in the hippocampus Expression of HSPB1,CAMS AP2,and SNCG.Results:1.Neurological deficit score: It can be seen that there was no neurological deficit in the sham operation group.The neurological deficit scores of the electroacupuncture group and the model group were significantly higher than those of the sham operation group on the 1st and 7th day after surgery(P<0.05)There was no statistically significant difference in the scores between the electroacupuncture group and the model group on the first day after the operation,and the scores of the electroacupuncture group were significantly improved compared with the m Podel group on the 7th day after the operation(P<0.05).Compared with the first day after the neurological deficit score,the score of the model group increased,the difference was statistically significant(P<0.05),and the score of the electroacupuncture group decreased significantly(P<0.05).2.Balance beam experiment: The balance ability of the sham operation group was evaluated normally,and the balance ability of the model group was significantly lower than that of the sham operation group on the 1st and 7th day after operation,that is,the neurological deficit score was higher than that of the sham operation group(P<0.05);There was no significant difference between the model group and the electroacu puncture group on the first day after the operation,and the score of the electroacupuncture group was significantly lower than that of the model group on the 7th day after the operation(P<0.05).3.Morris water maze experiment: There is no significant difference between the electroacupuncture group and the sham operation group;the escape latency of the model group is longer than that of the sham operation group(P<0.05);the electroacupuncture group and the model group are at the 3rd and 5th postoperatively,6 days shortened(P<0.05).In the space exploration experiment,there was no difference in the number of platform crossings between the electroacupuncture group and the sham operation group;the number of crossing platforms in the model group was significantly less than that of the sham operation group and the electroacupuncture group(P<0.05).4.TTC staining: The brain slices of the rats in the sham operation group are bright red,the rats in the model group can see large areas of gray-white infarcts,and the electro-acupuncture group can also see the gray-white infarcts,but the area is smaller than that of the model group.5.Fluorescence quantitative PCR experiment: Compared with the sham operation group,the expression of CACNα1d m RNA in the model group was significantly reduced,and the difference was statistically significant(P<0.05).The expression of HSPB1,CAMSAP2,and SNCG in the model group was significantly increased,and the difference was statistically significant.Significance(P<0.05);Compared with the electroacupu ncture group,the CACNα1d mRNA expression in the model group was reduced,and the difference was statistically significant(P<0.05),and the mRNA expression of HSPB1,CAMSAP2,and SNCG increased,and the difference was statistically significant(P<0.05).6.Western blotting experiment: Compared with the sham operation group,the protein expression of HSPB1,CAMSAP2,and SNCG in the model group increased,and the difference was statistically significant(P<0.05);compared with the electroacupuncture group,the model group’s The protein expression of HSPB1,CAMSAP2,and SNCG increased significantly,and the difference was statistically significant(P<0.05).7.Observation by immunohistochemical method: Compared with the sham operation group,the CACNα1d cell positivity of the model group was reduced,and the difference was statistically significant(P<0.05);compared with the electroacupuncture group,the model group HSPB1,CAMSAP2,SNCG The positive rate of cells increased(P<0.05).Conclusion:1.Acupuncture at Shenting and Baihui points can effectively promote the recovery of neurological function in rats with cerebral ischemia reperfusion.2.Acupuncture at Shenting and Baihui points can promote the learning and memory ability of rats with cerebral ischemia reperfusion and reduce neuronal damage.3.The mechanism of electroacupuncture to improve the learning and memory ability of MCAO rats may be by influencing the CACNα1D-HSPB1-CAMSAP2-SNCG path way in the hippocampus,promoting the expression of CACNα1d and inhibiting the expression of CAMSAP2,HSPB1 and SNCG.It is possible to repair brain function by promoting signal transduction,inhibiting the release of inflammatory transmitters,repairing vascular regeneration and reducing cell mitosis,reducing brain tissue damage,and improving learning and memory capabilities. |
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