| Objective:This study was conducted to observe the effects of transcutanclus electrical acupoint stimulation(TEAS)on the histomorphology of CA1 region of hippocampus in rats with chronic fatigue syndrome(CFS)and to investigate the possible mechanism of TEAS to improve learning The possible mechanism of TEAS to improve the learning and memory ability of CFS rats.Methods:Male Wistar rats were randomly divided into normal group,model group and TEAS group,10 rats in each group.The TEAS group was connected to a Han’s acupoint nerve stimulator at the "Zusanli"(ST36)and "Shenshu"(BL23)on both sides of the rats,with a dense and sparse waveform at 2 Hz/15 Hz and an intensity of 1-2 m A.The treatment was performed once a day for 20 min for 6 d,with 1 d of rest,for 4 weeks;the blank group and the model group were grasped and fixed under the same conditions without any treatment.The general conditions of the rats in each group were evaluated using the "Semi-quantitative Observation Scale for General Conditions of Experimental Rats",and the learning and memory abilities of the rats were observed by Morris water maze experiment after the intervention.The relative expressions of ERK,CREB and BDNF m RNA in hippocampal tissues were detected by real-time fluorescence quantitative PCR,and the relative expressions of ERK,CREB and BDNF protein in hippocampal tissues after the intervention were detected by immunoprotein blotting.Results:Comparison of the semi-quantitative observation table of general condition score of rats during modeling and treatment: before modeling,the rats in each group were in good health,and the general condition score was 0;during modeling,the rats in the modeling group gradually showed reduced confrontation,loose skin,thin and dull hair,and dilute stool;after modeling(22 d),the score of rats in the modeling group was significantly higher than that of the normal group(P < 0.05),and the difference between the two groups was not The difference between the two groups was not statistically significant(P > 0.05).At the end of modeling,all rats in the modeling group scored more than 4,suggesting that the CFS rat model was successfully established by the multifactorial compound stimulation method.During the treatment,all rats in the modeling group scored higher than 4 compared with the simultaneous point normal group(P < 0.01);compared with the simultaneous point modeling group,the rats in the TEAS group scored in the decreasing and tending to the normal group,and the score was lower than 4 after treatment(28 d)(P < 0.05).Compared with the pre-treatment group,the mental status,skin and hair nutrition degree,ear and tail color and fecal traits of the rats in the TEAS group showed some improvement,indicating that TEAS intervention could alleviate the fatigue status of CFS rats.Effects of transcutaneous electrical stimulation of "Zusanli"(ST36)and "Shenshu"(BL23)on learning and memory ability: After the intervention,the evasion latency of rats in the three groups showed a decreasing trend with the increase of Morris water maze training from day 1 to 4.Compared with the simultaneous point normal group,the evasion latency of rats in the model group was prolonged on days 1,2 and 3(P < 0.05),significantly prolonged on day 4(P < 0.01),and its number of crossing the platform was reduced on day 5(P < 0.05);compared with the model group,the evasion latency of rats in the TEAS group was shortened on days 2 and 3(P < 0.05),significantly shortened on days 1 and 4(P < 0.01),and the number of crossing the platform increased on day 5(P < 0.05).Effect of transcutaneous electrical stimulation of "Zusanli"(ST36)and "Shenshu"(BL23)on the pathology of CA1 area in hippocampal tissue:The results of hematoxylin-eosin staining after the intervention showed that the neuronal cells in the CA1 area of hippocampus of rats in the normal group were closely arranged,with oval or round nuclei,uniform cytoplasmic staining,clear and centered nuclei and clear edges;the number of neurons in the hippocampus of rats in the model group was reduced,the neuronal cells were arranged and scattered,some of the nuclei had lost their structure,the nuclei were fixed,the cytoplasm was unclear,the nuclei were inconspicuous,the position was irregular,the morphology varied,and even Some nerve cells showed obvious deformation and necrosis,the number of apoptotic cells increased significantly,and the neurons were wrinkled;in the TEAS group,the hippocampal neurons were still scattered,but the cell morphology was improved to some extent,the structure was clear,a small number of abnormal morphology and disorganized arrangement,some nuclei showed nuclear fixation,but the overall cell morphology was more regular,and the number of apoptotic cells was reduced.The effect of transcutaneous electrical stimulation of "Zusanli"(ST36)and "Shenshu"(BL23)on the ultrastructure of CA1 area of hippocampal tissue: The results of ultrasonic electron microscopy after the intervention showed that the neuronal nuclei in CA1 area of hippocampus of rats in the normal group had normal ovoid morphology,uniform distribution of chromatin in the nucleus,uniform cytoplasm,intact cell membrane and intact cristae;the neuronal nuclei in CA1 area of hippocampus of rats in the model group had blurred morphology,wrinkled cell cytoplasm,nuclear heterochromatin,and unevenly arranged nuclei.In the TEAS group,the neuronal nuclei in the CA1 area of hippocampus were normal,the chromatin was evenly distributed,the cytoplasm was uniform,the cell membrane was folded and the cristae were intact.Effects of transcutaneous electrical stimulation of "Zusanli"(ST36)and "Shenshu"(BL23)on ERK,CREB and BDNF gene expression in hippocampal tissue: Real-time fluorescence quantitative PCR results after intervention showed that the relative expression of ERK,CREB and BDNF in hippocampal tissue was significantly lower in the model group compared with the normal group(P<0.05,P<0.01);compared with the model group,the hippocampal The relative expressions of ERK,CREB and BDNF in the tissues of TEAS group were significantly higher(P<0.01,P<0.05).Effects of transcutaneous electrical stimulation of "Zusanli"(ST36)and "Shenshu"(BL23)on the expression of ERK,CREB and BDNF proteins in hippocampal tissues: Immunoprotein blotting results after the intervention showed that the relative expressions of ERK,CREB and BDNF proteins in hippocampal tissues were significantly lower in the model group compared with the normal group(P<0.05,P<0.01);compared with the model group,the relative expressions of ERK,CREB and BDNF proteins in hippocampal tissues in the TEAS group were significantly lower(P<0.05,P<0.01).The relative expressions of ERK,CREB and BDNF proteins were significantly higher in hippocampal tissues of the TEAS group compared with the model group(P<0.01,P<0.05).Conclusions:Transcutaneous electrical acupoint stimulation can improve the learning memory ability of CFS rats,and its mechanism of action may be related to improving the tissue structure of hippocampal CA1 region and upregulating ERK/CREB/BDNF expression. |
PDF Full Text Request