Based On MTOR Signaling Pathway, The Effects Of Moxibustion And Smog On Cognitive Impairment Of APP/PS1 Double Transgenic Mice Were Investigated

Objective:Targeting the mTOR signal pathway to determine the effects and further investigating the mechanism of moxibustion and moxa smoke on cognitive and non-cognitive behavior,oxidative status,A(3 and tau in APP/PS1 male mice.Method:Twelve C57BL/6 normal mice served as normal control while 36 APP/PS1 male mice were randomly divided into 3 groups(n=12/group),traditional moxibustion group,moxa smoke group,no-treatment control.Treatments were given 15 min,6 days a week.Mice in normal control and no-treatment control group were gently put in a fixator everyday.The traditional moxibustion mice were treated moxibustion on Guanyuan(CV4),while the mice in moxa smoke group were exposed to moxa smoke at the concentration range within 10-15mg/m3.After the 8 weeks treatment,the behavioral tests(Open field test,Novel object recognition task,Passive avoidance test,Morris water maze)were given to assess the level of neuropsychiatric impairment.Then subsequently all mice were sacrificed.Immunohistochem-ical staining was used to evaluate the A(31-42,senile plaques were detected by Thioflavine-S staining.GSH,SOD and MDA were determined in brain in all mice.Western blot method was used to evaluate the expression of p-tau(Ser202/Thr231)/tau,p-mTOR/mTOR,PI3K,p-AMPKα/AMPKα,PGC-1α,p-4E-BP1/4E-BPI,p-p70S6K/p70S6 K,p-ULK,LC3-I in hippocampus.The mRNA of mTOR,Akt and AMPKα1 were measured by RT-PCR.Results:1.Behavioral testComparing to no-treatment control,moxibustion group had significant higher number of rearings(p=0.048),moxa smoke group had significant higher moving distance(p=0.047)and number of crossing central zone in open field test(p=0.014).In novel object recognition task,moxibustion group spent more time exploring the novel object(p=0.012)and the frequency of exploring the novel object was higher in moxa smoke group(p=0.019).In passive avoidance test,moxibustion(p=0.039)and moxa smoke(p=0.000)groups showed lower frequency of mistakes compared with no-treatment control.All mice in 4 groups tended to perform better in Morris water maze with the training went on.Comparing to no-treatment control,the latency time of moxibustion was significant decreased in 4d(p=0.006).2.Aβ deposits and tau phosphorylationMice in no-treatment control expressed a lot of aggregation of Aβ1-42 in CA1 and CA3 of hippocampus.Less ggregation both in moxibustion group and moxa smoke group.Comparing to no-treatment control,the number of senile plaques were significant dereased(p=0.025)in moxibustion group.The mice in moxibustion(p=0.008)and moxa smoke(p=0.039)groups also had lower plaque burden.The expression of p-tau(Thr231)was significant decreased in moxibustion group(p=0.04).3.Oxidative status of brainComparing to normal control group,mice in no-treatment control had significant lower level of SOD(p=0.000)and higher level of MDA(p=0.009)in brain.Moxibustion decreased the production of MDA(p=0.009)and increased the level of SOD(p=0.02)and GSH(p=0.002).Moxa smoke group had a significant lower level of MDA(p=0.037).4.The upstream pathway of mTORComparing to normal control group,mice in no-treatment control had significant higher level of mTOR,p-mTOR and p-Akt/Akt,significant lower level of p-AMPKa,p-AMPKα/AMPKα.Comparing to no-treatment control,moxibustion group had significant lower level of mTOR(p=0.029),p-mTOR(p=0.005),p-mTOR/mTOR(p=0.015)、p-Akt/Akt(p=0.002)and significant higher level p-AMPKα(p=0.049),p-AMPKα/AMPKα(p=0.034).The expression of mRNA for mTOR was significant decreased(p=0.000)and AMPKal was increased in no-treatment control when compared with normal control group(p=0.014).Comparing to normal control group,the expression of mRNA for AMPKal was significant decreased(p=0.001).5.The downstream pathway of mTORThere was no difference between 4 groups on the expression of 4E-BP1 and p-4E-BP1.Comparing to normal control group,mice in no-treatment control had significant higher expression of p-p70S6K/p70S6K(p=0.001)and p-ULK(p=0.028).Moxibustion group(p=0.003)and moxa smoke(p=0.006)group had significant lower of level p-p70S6K/p70S6K compared with no-treatment control.The expression of LC3-Ⅱ was also increased in moxibustion group.Conclusion:1.Moxibustion and moxa smoke showed different effect on the behavioral change of APP/PS1 mice.Moxibustion treatment is capable of enhancing the learning and memory,reducing the Aβ deposits and tau phosphorylation.Moxa smoke can modify the mood behavioral change and decreased the Aβ deposits.2.Moxibustion might delay the pathological process of Alzheimer by inhibiting the activation of Akt,activating AMPKa and then damping down mTOR,regulating protein translation,promoting autophagy.3.Moxa smoke might delay the pathological process of Alzheimer by inhibiting the activation of Akt,mTOR and p70S6K.