Effect Of Xylazine On PKA?ERK1/2/CREB/BDNF Signaling Pathway In Brain Nervous System Of Rats

Male wistar rats were used for this study and xylazine anesthesia was given.Through measured the changes of PKA?ERK1/2/CREB/BDNF signaling pathways related protein and gene expression in different brain regions and different anesthesia period to investigate the mechanism of xylazine anesthesia.Methods:35 male wistar rats were divided into control group?C?and anesthesia group?X?.The anesthesia group was randomly divided into 6 groups?n=5?:Induction group?X1?,righting reflex disappeared 0 min group?X2?,righting reflex disappeared 30 min group?X3?,righting reflex disappeared 60 min group?X4?,righting reflex recovery 0 min group?X5?and righting reflex recovery 30 min group?X6?.After the control group was intraperitoneally injected normal saline of equal volume for 8 min and after each time point of intraperitoneal injection xylazine?70mg/kg?,the rats were sacrificed immediately and took brain tissue.The expression of GluR1,GluR2,ERK1/2,PKC,PKA,CREB,c-jun,c-fos,fos-B and BDNF mRNA were detected by qRT-PCR.The protein expression of GluR1,GluR2,ERK1/2/p-ERK1/2,PKA/p-PKA,PKC/p-PKC,CREB/p-CREB and BDNF was detected by western blot.The activity of ATPase was measured by ATPase kit and the content of cAMP was measured by ELISA kit.Through the changes of these index to investigated the effect of xylazine anesthesia in different brain regions and neurons in rats and the relationship between PKA?ERK1/2/CREB/BDNF signaling pathways.Results:?1?Tissue qRT-PCR results showed that compared with control group,the level of GluR1mRNA decreased significantly in the cerebrum and hippocampus;the level of GluR2 and ERK1/2mRNA decreased significantly in the cerebrum;the level of PKC mRNA did not change significantly in all brain regions;the level of PKA mRNA decreased significantly in the hippocampus and thalamus;the level of CREB mRNA decreased significantly in the thalamus and brainstem.the level of c-jun,fos-B,c-fos and BDNF mRNA decreased significantly in all brain regions?P<0.05?.?2?Cell qRT-PCR results showed that the expression of ERK1/2 mRNA was significantly different between K group and ERK1/2i group compared with C group?P<0.05?.The expression of PKA mRNA was significantly different between K group and PKAi group compared with C group?P<0.05?.The expression of ERK1/2 mRNA no significant difference each group compared with the C group?P>0.05?.The expression levels of c-jun,c-fos,fos-B and BDNF mRNA were significantly different between the K group,the ERK1/2i group and the PKAi group compared with the C group?P<0.05?.?3?Tissue western blot result showed that compared with C group,the protein expression of GluR1,p-ERK1/2 and p-PKA was decreased significantly in each brain area compared with the control group;GluR2 decreased significantly in the brain,cerebellum,thalamus,and brain stem;ERK1/2 decreased significantly in the cerebrum,cerebellum and brainstem;PKA decreased significantly in the hippocampus and thalamus;PKC and p-PKC were not significantly different in each brain area;BDNF decreased significantly in all brain regions?P<0.05?.?4?Cell western blot result showed that compared with the control group,the protein expression of GluR1,p-ERK1/2 and p-PKA was decreased significantly in each brain area.GluR2significantly decreased in the cerebrum,cerebellum,thalamus,and brainstem;ERK1/2 decreased significantly in the cerebrum,cerebellum and brainstem;PKA significantly decreased in the hippocampus and thalamus;PKC and p-PKC were not significantly different in each brain area;BDNF decreased significantly in all brain regions?P<0.05?.?5?The activity of Na⁺-K⁺-ATPase was decreased significantly in the cerebrum,hippocampus,thalamus and brainstem?P<0.05?.?6?The activity of Ca²⁺-Mg²⁺-ATPase was significantly decreased in each brain regions?P<0.05?.?7?The content of cAMP decreased significantly in each brain regions?P<0.05?.Conclusion:?1?Xylazine make the body in anesthesia by affecting neuronal system AMPA membrane receptors,CREB protein pathways and c-fos?fos-B?c-jun and BDNF gene transcription.?2?ERK1/2 and PKA can regulate the expression of CREB,and affect the downstream indicators of CREB,indicating that xylazine anesthetic effect is by inhibiting the CREB pathway on rats.?3?The changes of ERK1/2/p-ERK1/2,PKA/p-PKA,CREB/p-CREB,c-jun,c-fos,fos-B and BDNF were most obvious in the cerebrum,cerebellum and hippocampus.It shows that xylazine anesthesia on rats is caused by affecting the expression of the above related factors and the cascade reaction between them.