The Effect Of Acupuncture On Signal Molecule Of PACAP/PAC1-PKA-CREB Signaling Pathway Of The Bed Nucleus Of Stria Terminalis In Anxiety Mice

ObjectiveIn order to reveal the central PACAP mechanism of acupuncture anxiolytic,the study invesgated the effect of acupuncture on signal molecule of PACAP/PAC1-PKACREB signaling pathway in the bed nucleus of stria terminalis(BNST).MethodsThirty healthy,clean grade,male C57BL/6J mice were divided into three groups according to behaviors result,including blank group,model group,and acupuncture group.Each group contained 10 mice.Anxious mouse model of CUS was established based on the literature.The acupuncture group selected "Shenmen" and "Neiguan" for acupuncture treatment for 30 minutes,followed by twirling of needle every 5 minutes,30 seconds for each time,left and right alternately,once every other day,treatment continued for 15 days.Elevated plus maze test(EPM)was used to test the anxiety-like behaviors for each group.The plasma Corticosterone(CORT)level was determined by Enzyme Linked Immunosorbent Assay(ELISA).Immunohistochemistry was used to detect the expression levels of Pituitary adenylate cyclase activating polypeptide,(PACAP),PAC1,Protein kinase A(PKA),cAMP-responsive element binding protein(CREB)in BNST.Results 1.Behavioral test results(1)OT%: compared with the blank group,OT% in the model group decreased significantly(P<0.05);OT% in the acupuncture group was significantly higher than that in the model group(P<0.05),and its level is same to the blank group.(2)OE%: Compared with the model group,OE% of the acupuncture group was higher than the model group significantly(P<0.05).2.HPA axis related indicators Plasma CORT,after CUS the plasma CORT levels in model group increased than blank group significantly(P<0.01);the plasma CORT levels decreased significantly after acupuncture treatment(P<0.01)3.Molecular expression of PACAP/PAC1-PAC1-PKA-CREB signaling pathway in BNST(1)PACAP: The expression of it in the BNST for anxious model mice was significantly reduced compared with the blank group(P<0.01);compared with the model group,the acupuncture can significantly increased PACAP expression in BNST(P<0.01).(2)PAC1: the expression of PAC1 in BNST has no significant difference among three groups(P>0.05).(3)PKA: The expression of it for the anxiety model mice in BNST was significantly decreased compared with the blank group(P<0.01).Compared with the model group,acupuncture could significantly increase the expression of PKA in the BNST,and the comparison between groups was statistically significant.(P<0.05).(4)CREB: The expression of it in the BNST for mice after modeling was significantly decreased compared with the blank group(P<0.01);acupuncture could increase the expression of CREB in the BNST,which was statistically significant compared with the model group(P<0.05).Conclusion 1.Acupuncture at "Shenmeng" and "Neiguan" points have a good anxiolytic effect on anxiety mice induced by CUS.2.Activation of PACAP/PAC1-PKA-CREB signal pathway in the BNST and reducing the plasma CORT level in anxious mice induced by CUS may play an important role in anti-anxiety for acupuncture.