| Objective:Rapid aging dementia mice(SAMP8)and their homologous anti-aging mice(Senescence-accelerated resistant mouse/1,SAMR1)were used to observe the effect of well point bloodletting on FNDC5/Irisin in hippocampal CA1 region,to explore whether bloodletting can have a therapeutic effect on AD,whether FNDC5/Irisin in hippocampal CA1 region is one of the mechanisms of bloodletting in the treatment of AD,and the possible source of elevated iris in hippocampus.Methods:Experiment 1: study on the effect of bloodletting on the behavior of rapidly aging dementia mice.The purpose of this part of the experiment is to construct the animal experimental platform of twelve well acupoints for the treatment of senile dementia,and to determine the therapeutic effect of bloodletting on cognitive impairment in rapidly aging dementia mice(SAMP8).Sixteen SAMP8 mice and 9 SAMR1 mice were randomly divided into two groups: model group and bloodletting group,SAMR1 mice in the control group without any treatment(no binding fixation,no well acupoint bloodletting intervention),the model group received sham acupuncture at the well acupoint every day(no skin breaking,no bleeding),and the bloodletting group was punctured and bled at the twelve well acupoints on both forelimbs every morning,once a day,for two weeks.On the second day after treatment,the open field test and Morris water maze test were performed to observe the effects of various interventions on the behavioral indexes of mice.In experiment 2:the effect of bloodletting on the expression of iris in the hippocampus of rapidly aging dementia mice.Sixteen SAMP8 mice were randomly divided into two groups: model group(n = 8),bloodletting group(n = 8)and control group(n = 9).The methods of treatment and intervention were the same as those in experiment 1.After two weeks of intervention,the behavior test was carried out.After the behavior test,the hippocampal tissue of mice was taken,and the heart blood was taken.The content of serum iris was detected by ELISA,the expression of iris in mouse hippocampal tissue was detected by western blotting,and the expression of iris m RNA in mouse hippocampal tissue was detected by real time reverse transcription PCR.Experiment 3: study on the source of iris in peripheral blood of rapidly aging dementia mice.Twelve SAMP8 mice were randomly divided into model group(n = 6)and bloodletting group(n = 6).SAMR1 mice were used as control group.The intervention method was the same as that of experiment 1.Muscle and adipose tissue were taken after two weeks of intervention.The expression of iris in muscle and adipose tissue of mice was detected by Western blot and the expression of iris m RNA in muscle and adipose tissue was detected by RT-PCR.Results:In experiment 1,the animal experimental platform of twelve well acupoints bloodletting in the treatment of Alzheimer’s disease was successfully constructed.Open field test(1)comparison of the total distance: the total distance of the control group was higher than that of the model group,P < 0.06,and there was significant difference between the bloodletting group and the model group(P < 0.05).(2)the distance of central movement: compared with the control group,the distance of central movement in the model group was less than that in the control group(P < 0.05),and the distance of central activity in the bloodletting group was more than that in the model group(P < 0.05).There was statistical significance(P < 0.05).(3)Central activity time: the central activity time in the model group was less than that in the other two groups,but there was no significant difference among the three groups(P > 0.05).Morris water maze(1)location navigation experiment: compared with the control group,the escape latency of the model group was significantly longer than that of the control group,and there was a significant difference between the two groups on the 5th day(P < 0.001).Compared with the model group,the escape latency of the bloodletting group was significantly shorter than that of the model group.(2)Spatial exploration: compared with the control group,the number of times of crossing the platform in the model group was significantly less,and there was significant difference between the model group and the control group(P < 0.05).Compared with the model group,the times of crossing the platform in the bloodletting group increased,and there was significant difference between the two groups(P < 0.05).In experiment 2,the level of serum iris in mice was detected by ELISA: compared with the control group,the level of serum iris in the model group decreased,and that in the bloodletting group increased compared with the model group(P < 0.05).The level of iris in hippocampus of mice was detected by Western blot: compared with the control group,the ratio of iris / tubulin in the model group was significantly higher than that in the control group,and the ratio of iris / tubulin in the model group was significantly higher than that in the model group,and there was significant difference between the bloodletting group and the model group.The expression level of Iris gene in hippocampus of mice was determined by RT-PCR: there was no significant difference among the three groups(P > 0.05).In experiment 3,the iris level in muscle and adipose tissue of mice was detected by Western blot: there was no difference in the expression of iris protein in muscle and adipose tissue among the three groups,and the expression level of iris gene in muscle and adipose tissue was determined by RT-PCR.There was no difference in the expression of iris m RNA in muscle and adipose tissue among the three groups.Conclusion:1.Bloodletting can improve the spatial learning and memory ability and other dementia behaviors of rapidly aging dementia mice(SAMP8).2.Bloodletting therapy can increase the content of Iris in serum and the expression of Iris protein in hippocampus,suggesting that the increase of Iris in hippocampus may come from peripheral blood after bloodletting.3.The elevated Iris in the hippocampus may come from muscle. |
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