Effects Of Tiletamine And Tramadol On The LKB1-AMPK-mTOR Signaling Pathway In Rats' Brain

XFM is a special compound anesthetic for miniature pigs with short time induced,anesthesia induction and anesthesia duration suitable and less side effects developed by our team.Tiletamine and tramadol as one of the components of XFM,are acting on the animal's central nervous system(CNS),and have a real analgesic effect.Recent studies have shown that the key proteins in the LKB1-AMPK-m TOR signaling pathway are involved in the regulation of pain and neuronal excitability,suggesting that the LKB1-AMPK-m TOR signaling pathway may be involved in the regulation of analgesic.Therefore,in this study,rats were injected intraperitoneally with normal saline,tiletamine hydrochloride and tramadol hydrochloride,and using quantitative real-time polymerase chain reaction and western blot analysis to detect key protein gene transcription and protein phosphorylation levels,respectively in the LKB1-AMPK-m TOR signaling pathway in rats central nervous system(cerebral cortex,hippocampus,thalamus,cerebellum,brainstem)at different times after injection,to explore their analgesic mechanism,to speculate on XFM general anesthesia analgesic mechanism.54 Sprague-Dawley rats were divided into control(n=6),anesthesia group(n=24)and analgesia group(n=24).After rats were injected intraperitoneally with saline,tiletamine and tramadol,they were executed at 10,20,40 or 60 min post injection.The cerebral cortex,hippocampus,thalamus,cerebellum and brainstem were immediately taken out to evaluate the m RNA and protein phosphorylation levels of liver kinase B1(LKB1),AMP-activated protein kinase(AMPK)and e IF4E-binding protein 1(4EBP1)using quantitative real-time polymerase chain reaction and western blot analysis.The results are as follows:(1)Under the intervention of the drug,the relative transcription of LKB1 mRNA,AMPK?1m RNA and AMPK?2 m RNA was significantly increased(p<0.01),while the relative transcription of 4EBP1 mRNA was inhibited(p<0.01).(2)Tiletamine and tramadol induced the phosphorylation of LKB1 and AMPK(p<0.01),whereas the phosphorylation level of 4EBP1 was inhibited(p<0.01).(3)In the central nervous system of rats,tiletamine and tramadol showed an effective regulatory effect in the hippocampus,thalamus,cerebellum and brainstem with 20 min and 40 min post injection.In summary,the findings indicate that the LKB1-AMPK-m TOR signaling pathway is involved in anesthetic analgesia regulation of tiletamine and tramadol,whereas the hippocampus,cerebellum,thalamus and brainstem of rats may be LKB1-AMPK-m TOR signaling pathways involved in anesthesia and analgesic targets.The activation of the LKB1-AMPK-m TOR signaling pathway on the corresponding target protein of the drug produces anesthetic analgesic effect that may be the mechanism of tiletamine and tramadol,which also provide a solid basis for further revealing the XFM general anesthesia analgesic mechanism.