| Objective:1.To explore whether acupuncture can improve cognitive function in CFS rats.2.The effect of acupuncture on the improvement of cognitive dysfunction in CFS model rats is achieved through the regulation of the miRNA-184/ 5-HT/Gnai/PLC/Casp-3 pathway and the inhibition of the apoptosis of the hippocampus cells.Method:1.Experiment 1: effect of acupuncture on cognitive impairment in CFS rats60 2 month old male SD rats were randomly divided into normal group(group A),model group(group B),acupuncture group(group C)and sham acupuncture group(D group).Rats in group B,C and D were trained for 21 days by exhaustive swimming plus chronic restraint.After the completion of the model,group A was not treated with any treatment.Group B began to use self-made fixtures to fix rats on third days after the model was successful,fixed time for 15 minutes,once a day,after 5 days of continuous treatment for 2 days for 21 days.Group C was given acupuncture "Baihui","Shenshu","Shenshu","Shenshu","spleen Yu",After three li treatment,the time frequency was the same as that in group B;group D in group C was treated with acupuncture at 5mm point beside the acupuncture point,and the time frequency was the same as that in group C.The cognitive function of CFS rats was detected by elevated maze,open field test and water maze test after modeling and after treatment.2.Experiment two: the regulation mechanism of miRNA-184/ 5-HT/Gnai/PLC/Casp-3 pathway in rats with CFS cognitive dysfunction induced by Acupuncture70 2-month-old male SD rats were randomly divided into normal group(group E),model group(group F),acupuncture group(group G),saline injection +acupuncture group(group H)and miRNA-184 antagonist injection + acupuncture group(group I).The rats of F,G,H,I group were made by exhaustion swimming and chronic binding method for 21 days.The group was given acupuncture of "Baihui","Shen gate","Shenshu","Shenshu","spleen Yu","after three li" treatment,once a day,after 5 days of continuous treatment for 2 days,a total of 21 days.After the treatment of acupuncture and moxibustion,the survival rate of hippocampal cells was measuredby Nissl staining.The content of miR-184 in the hippocampus was detected by RT-PCR,the content of 5-HT in hippocampus was detected by ELISA,and the content of Gnail protein and PLC protein in hippocampus was measured by Western Blot method,and the content of Casp-3 protein in hippocampus was determined by immunohistochemistry.Result:1.Experiment 1: effect of acupuncture on cognitive impairment in CFS rats(1)The elevated cross maze: after the success of the CFS model,compared with the A group,the opening time of the B,C and D groups was significantly shortened and the number of entry arms decreased significantly.The difference was statistically significant(P<0.01).After the acupuncture treatment,the opening time of the C group was significantly longer than that of the B group,and the number of entry arms increased significantly.The difference was significantly increased.It was statistically significant(all P<0.01).Compared with group C,the opening time of open arm in group D was significantly shortened,and the number of entry arms decreased significantly,and the difference was statistically significant(P<0.01).(2)Open field experiment: after the success of CFS modeling,compared with the A group,the number of cross lattice of B,C and D was significantly reduced and the residence time of the central lattice was obviously shortened.The difference was statistically significant(P<0.01).Compared with the B group,the number of trans lattice times in the C group was significantly increased and the residence time of the central lattice was significantly prolonged,and the difference was statistically significant after the acupuncture treatment.Study significance(all P<0.01);compared with group C,the number of cross grid times in group D decreased significantly and the residence time of central lattice shortened obviously,and the difference was statistically significant(P<0.01).(3)The water maze experiment: after the success of the CFS model,compared with the A group,the water maze in the B,C and D groups increased significantly,the retention time of the effective area was significantly reduced,and the difference was statistically significant(P<0.01).After the acupuncture treatment,the water maze of the C group was significantly reduced and the residence time of the central lattice increased significantly compared with the B group,and the difference has a significant difference.There were statistical significance(all P<0.01).Compared withgroup C,the water maze of group D increased significantly and the residence time of central lattice decreased significantly,and the difference was statistically significant(P<0.01).2.Experiment two: the regulation mechanism of miRNA-184/ 5-HT/Gnai/PLC/Casp-3 pathway in rats with CFS cognitive dysfunction induced by Acupuncture(1)The survival rate of hippocampal cells: after the success of the model,compared with the E group,the apoptosis of hippocampal cells increased significantly in group F,and the difference was statistically significant(P<0.05).Compared with the F group,the apoptosis of hippocampal cells in the G group was significantly decreased,the difference was statistically significant(P<0.05).Compared with the I group,the hippocampal cells in the H group were withered.The death rate was significantly reduced,the difference was statistically significant(P<0.05).(2)The content of Casp-3 protein in hippocampus: after the model was successful,the content of Casp-3 protein in hippocampus of F group decreased significantly compared with that of group E,and the difference was statistically significant(P<0.05).After acupuncture treatment,the content of Casp-3 protein in hippocampus of G group was significantly higher than that of F group,and the difference was statistically significant(P<0.05);compared with I group,H group rats were compared.The content of Casp-3 protein in hippocampus increased significantly,and the difference was statistically significant(P<0.05).(3)The content of PLC protein in hippocampus: after the model was successful,the content of PLC protein in hippocampus of F group decreased significantly compared with E group,and the difference was statistically significant(P<0.05).After acupuncture treatment,the content of PLC protein in hippocampus of G group was significantly higher than that of F group,and the difference was statistically significant(P< 0.05).Compared with I group,H group rat hippocampal PLC Protein content increased significantly,the difference was statistically significant(P<0.05).(4)Hippocampal Gnail protein content: after model success,compared with group E,the content of Gnail protein in hippocampus of F group decreased significantly(P<0.05).After acupuncture treatment,the content of Gnail protein in hippocampal Gnail in G group was significantly higher than that in F group,and the difference was statistically significant(P<0.05);compared with I group,H group rat sea was compared.The content of Gnai protein in horses increased significantly,and thedifference was statistically significant(P<0.05).(5)Hippocampal 5-HT content: after the successful modeling,compared with E,the5-HT content in the hippocampus of F group decreased significantly,the difference was statistically significant(P<0.05).After acupuncture treatment,the hippocampal5-HT in G group was significantly higher than that in F group,and the difference was statistically significant(P<0.05).Compared with the I group,the hippocampus 5-HT content in H group was significantly higher than that in I group.The difference was statistically significant(P<0.05).(6)miRNA-184 expression in hippocampus: after successful modeling,the expression of miRNA-184 in hippocampus of F group decreased significantly compared with group E,and the difference was statistically significant(P<0.05).After acupuncture treatment,the expression of miRNA-184 in hippocampal miRNA-184 in G group was significantly enhanced compared with F group,and the difference was statistically significant(P <0.05).Compared with I group,the H group rat hippocampus The expression of iRNA-184 increased significantly,and the difference was statistically significant(P<0.05).Conclusion:1.CFS model rats have cognitive impairment,which shows a significant decline in the ability to explore,the ability of self-activity,and the ability of learning and memory.Compared with the blank control and false acupuncture,acupuncture can effectively improve the cognitive dysfunction of the CFS model rats.2.The regulation of cognitive dysfunction in CFS model rats by acupuncture may be achieved by up regulating the expression of miR-184 in the hippocampus,activating the 5-HT/Gnai/PLC pathway in the hippocampus,inhibiting the expression of Casp-3protein and inhibiting the apoptosis of hippocampal neurons. |
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