| As an experimental animal model, minipigs were widely used in the study of human anatomy, physiology, biochemistry, mechanism of disease, and xenotransplant. It is necessary to provide the satisfactory anaesthesia and analgesia for proceeding of the correlated territory smoothly. So the XFM and specificityantagonist were successfully prepared by this team. In the last century, the lipid theory of anaesthesia was accounts with an important position. But with the deepening development of anaesthesia mechanism, and the development and improvement of protein theory and gene theory, studies reported that expression of some quick express gene changed after anaesthesia. And these studies prompted that there were some positive relationship between the potency of anaesthetic and genetic variation. At the same times, studies found that GABA receptors and its encoding genes had a close relationship with general anesthesia. Therefore the study of anaesthesia and wakeup mechanism, from the perspective of protein and gene, became more and more important.PI3K/Akt- P70S6 K signaling pathway is a classical information transfer path. In this pathway, the key member, Akt, is the key point of the survivorship after the cell injury. Although the studies of PI3K/Akt signaling pathway were concentrate upon the protection of the injure after the postischemia of organ and tissue, PI3 K can not only affect the levels of intracellular Ca2+, but also be activated by the α2 adrenergic receptor indirectly. In order to understand the central action of the XFM and its specificity antagonist, and to guide the clinical medication, in this study, the relationship between PI3K/Akt signaling pathway and XFM specificity antagonist were researched and interaction mechanism of XFM and its specificity antagonist may be identified.In this study, 78 wistar rats were divided into Anaesthesia Group(AG group, n=30), Wake Group(W group, n=18), and(Anaesthesia-Wake group, n=30). Animals in AG group were randomly divided into five subgroups: control group I(CG1 group), abolition of righting reflex instantly after anaesthesia(AG1 group), 1 h after righting reflex instantly after anaesthesia(AG2 group), recovery instantly after anaesthesia(AG3 group), recovery 1 h after anaesthesia(AG4 group); animals in W group, were randomly divided into three subgroups: the control group II(CG2), 5 min after administration of antagonist(W1 group), and 1 h after administration of antagonist(W2 group); animals in AW group were randomly divided in to five subgroups: the control group III(CG3 group), recovery instantly when wake at the time of abolition of righting reflex instantly after anaesthesia(AW1 group), recovery 1 h when wake at the time of abolition of righting reflex instantly after anaesthesia(AW2 group), recovery instantly when wake at the time of 1 h after righting reflex instantly after anaesthesia(AW3 group), recovery 1 h when wake at the time of 1 h after righting reflex instantly after anaesthesia(AW4 group). Rats in the experiment groups were administrated XFM or specificity antagonist for XFM, samples were collected at different timepoints, and after the treatment of sample, the transcriptional levels of PI3K/Akt m RNA, m TOR, and P70S6 K were detected by realtime fluores-cence quantitative PCR, and the expression of protein were detected by Western Blot. The effects of XFM and its specificity antagonist on the PI3K/Akt signaling pathway m RNA and protein in rats’ brain were studied. The results were shown that:1. Effects of XFM and its specificity antagonist on the expression of PI3 K protein and transcription of PI3 K m RNAAfter the administration of XFM, the expression of PI3 K protein and the transcription of PI3 K m RNA in cerebral cortex, hippocampus, and thalamus were activated significantly, and the change of protein and m RNA were at equal pace. Injection of the specificity antagonist for XFM alone could significantly inhibit the expression of PI3 K protein and the transcription of PI3 K m RNA in cerebral cortex, hippocampus, and brainstem. Using the specificity antagonist for XFM to wake the rats anaesthetized with XFM may cause the significant inhibition of the expression of PI3 K protein and the transcription of PI3 K m RNA in cerebral cortex, hippocampus, and brainstem. It indicated that the anaesthesia effect of XFM may be relation with the activation of the expression of PI3 K protein and the transcription of PI3 K m RNA in definite encephalic region, and it also indicated that the awake efficacy of antagonist may be relation with the inversion of the activation of PI3 K caused by XFM.2. Effects of XFM and its specificity antagonist on the expression of Akt protein and transcription of Akt m RNAThe expression of PI3 K protein and the transcription of Akt m RNA in cerebral cortex, hippocampus, and brainstem increased significantly after administration of XFM, and there were some associativity change between the protein and m RNA. Administration of the specificity antagonist for XFM alone could significantly inhibit the expression of Akt protein and the transcription of Akt m RNA in hippocampus, and brainstem. Using the specificity antagonist for XFM to wake the rats anaesthetized with XFM may cause the significant inhibition of the expression of Akt protein and the transcription of Akt m RNA in cerebral cortex, hippocampus, and brainstem. It indicated that the anaesthesia effect of XFM may be relation with the activation of the expression of Akt protein and the transcription of Akt m RNA in definite encephalic region, and it also indicated that the awake efficacy of antagonist may be relation with the inversion of the activation of Akt caused by XFM.3. Effects of XFM and its specificity antagonist on the expression of m TOR protein and transcription of m TOR m RNAAfter the injection of XFM, the expression of m TOR protein and the transcription of m TOR m RNA in cerebral cortex, hippocampus were activated significantly, Injection of the specificity antagonist for XFM could significantly inhibit the expression of m TOR protein and the transcription of PI3 K m RNA in cerebral cortex, hippocampus, and brainstem. At the time of abolition of righting reflex, injected the specificity antagonist instantly to wake the rats anaesthetized with XFM, the expression of PI3 K protein and the transcription of PI3 K m RNA in cerebral cortex, hippocampus, and brainstem were significantly inhibited, and one hour after abolition of righting reflex, injected the specificity antagonist to wake the rats anaesthetized with XFM, the expression of m TOR protein and the transcription of m TOR m RNA in hippocampus and thalamus were significantly inhibited. It indicated that the anaesthesia effect of XFM may be relation with the activation of the expression of m TOR protein and the transcription of m TOR m RNA in definite encephalic region, and it also indicated that the awake efficacy of antagonist may be relation with the inversion of the activation of m TOR in definite encephalic region caused by XFM.4. Effects of XFM and its specificity antagonist on the expression of P70S6 K protein and transcription of P70S6 K m RNAThe expression of PI3 K protein and the transcription of P70S6 K m RNA in cerebral cortex, thalamus, and brainstem increased significantly after administration of XFM. Administration of the specificity antagonist for XFM could significantly inhibit the expression of P70S6 K protein and the transcription of P70S6 K m RNA in hippocampus and thalamus. Using the specificity antagonist for XFM to wake the rats anaesthetized with XFM may cause the significant inhibition of the expression of P70S6 K protein and the transcription of P70S6 K m RNA in cerebral cortex and thalamus. It indicated that the anaesthesia effect of XFM may be relation with the activation of the expression of P70S6 K protein and the transcription of P70S6 K m RNA in definite encephalic region, and it also indicated that the awake efficacy of antagonist may be relation with the inversion of the activation of P70S6 K.In conclusion:(1) There were some activation effects of XFM on the PI3K/Akt signaling pathway and its downstream signaling molecules, and this activation of XFM could be obviously shown in cerebral cortex, hippocampus, maybe this is one of the target position of PI3K/Akt signaling pathway, that participated in the anaesthesia effects of XFM.(2) Administration of the specificity antagonist for XFM alone could significantly inhibit the PI3K/Akt signaling pathway and its downstream signaling molecules. The inhibition effect of the antagonist could be obviously shown in thalamus and hippocampus, and it indicated that the performance of wake efficacy from the specificity antagonist may be achieved through the inhibition of PI3 K / Akt signaling pathway.(3) The effects of XFM on the PI3K/Akt signaling pathway were opposite with the specificity antagonist for XFM, and maybe this is one of interaction mechanism between XFM and its specificity antagonist. But the specificity antagonist can not completely antagonistic the activation effect of XFM on the PI3K/Akt signaling pathway and its downstream molecules, and it prompted that the study on the mechanism of general anaesthesia were complex and multidigit. |
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