| Objective:To observe the effect of Electroacupuncture on the learning and memory ability of middle cerebral artery occlusion(MCAO)model rats and the expression of key genes and proteins in the CaCN / Ras / ERK pathway in the hippocampus,and to explore the mechanism of electroacupuncture in improving the learning and memory ability,so as to treat vascular cognitive impairment Improvement(VCI)provides theoretical and experimental basis.Methods:24 male SD rats with a body mass of(280 ± 20)g were randomly divided into three groups: Electroacupuncture group,model group and sham operation group.The MCAO model of electroacupuncture group and model group was prepared with reference to longa modified thread bolt method.The other operations were the same as those in the model group.In electroacupuncture group,shenting point and Baihui Point were intervened by electroacupuncture.The frequency of electroacupuncture was 2-10 Hz and the voltage was1.5-2mA.The intensity gradually increased to the degree of slight shaking of the rat's ears without hoarseness.The needle was kept for 20 min.The treatment was started 24 hours after the operation,once a day for 7 days.The positive and negative poles of each electroacupuncture were different from the previous one.Model group and sham operation group were treated with the same conditions of grasping and binding as electroacupuncture group.On the first and seventh day after operation,neurobehavioral scores were made according to the Zea longa method;water maze test was conducted on the third to seventh day after operation;two rats in each group were randomly selected for 2,3,5-triphenyltetrazolium chloride after the end of electroacupuncture intervention on the seventh day after operation The expression of key genes(CaCN?1d,RasGRP,ERK)in the CaCN / Ras / ERK pathway was detected by real-time fluorescence quantitative polymerase chain reaction(PCR);the expression of key proteins(CaCN?1d,RasGRP,ERK)in the CaCN / Ras / ERK pathway was detected by Western blot(WB).Result:1.Neurological deficit score: it can be seen that there was no neurological deficit score in the sham operation group,and the neurological deficit score in the electroacupuncture group and the model group was significantly higher than that in the sham operation group on the first and seventh days after operation(P<0.01).There was no significant difference between the electroacupuncture group and the model group on the first day after operation,and the score in the electroacupuncture group was significantly improved on the seventh day after operation(P<0.01).Compared with the first day,the score of the model group increased significantly(P<0.05),while that of the electroacupuncture group decreased significantly(P<0.01).2.Morris water maze experiment: There was no significant difference between the electroacupuncture group and the sham operation group;the escape latency of the model group was longer than that of the sham operation group(P<0.05);the escape latency of the electroacupuncture group and the model group was shorter than that of the sham operation group on the 3rd,5th and 6th day after operation(P<0.05).There was no difference in the number of platform crossing between the electroacupuncture group and the sham operation group in the space exploration experiment;the number of platform crossing in the modelgroup was significantly lower than that in the sham operation group and electroacupuncture group(P<0.01).3.TTC staining: The brain slices of rats in the sham operation group showed bright red,large area of gray white infarction could be seen in the model group,and gray white infarction could be seen in the electroacupuncture group,but the area was smaller than that in the model group.4.Quantitative Real-time PCR test: Compared with the sham operation group,the mRNA expression of CaCN?1d,RasGRP and ERK in the electroacupuncture group was not statistically significant;compared with the model group,the mRNA expression of CaCN?1d and ERK in the electroacupuncture group was significantly lower,the difference was statistically significant(P<0.01),and the mRNA expression of RasGRP was increased,the difference was statistically significant(P<0.05).5.Western blotting test: Compared with the sham operation group,the protein expression of CaCN?1d and ERK in the model group increased,and the protein expression of RasGRP decreased;compared with the model group,the protein expression of CaCN?1d and ERK in the electroacupuncture group decreased,and the protein expression of RasGRP increased.Conclusion:1.Electroacupuncture at shenting and Baihui points can improve the learning and memory ability of MCAO rats2.The mechanism of electroacupuncture to improve the learning and memory ability of MCAO rats may be that it can inhibit the influx of calcium and decrease the phosphorylation of downstream cascade core units by affecting the CaCN / Ras / ERK pathway of hippocampus,so as to reduce the damage of cerebellum and improve the learning and memory ability. |
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