| Objective To explore the protective effect and mechanism of post-treatment with sevoflurane mediated by Akt/GSK-3β pathway on brain hypoxia/reoxygenation injury of zebrafish.Methods Zebrafish hypoxia/reoxygenation model was established,and healthy zebrafishes of the same size were divided into Control group(Control group),hypoxia/reoxygenation group(H/R),sevoflurane post-treatment group(Sevo group),hypoxia/reoxygenation +GSK-3β inhibitor group(H/R+TDZD-8)for experiments.The m RNA expressions of hypoxia-inducing factors Hif-1αa and Hif-1αb at different reoxygenation time points were determined by q RT-PCR,GC-MS was used to detect the concentration of sevoflurane in fish water and brain tissue homogenate,infarct size was detected by TTC staining,Ultrastructure of mitochondria in zebrafish neurons was detected by electron microscopy,and the glutathione levels in zebrafish brain tissue were measured,and the protein expression levels of HIF-1α,p-Akt,Akt,GSK-3β,p-GSK-3β(Ser 9)and MAP2 were detected by Western Blot,MAP2 expression was detected by immunofluorescence,and Behavioral detection of optokinetic response frequency in zebrafish.Results Compared with Control group,The m RNA levels of Hif-1αa and Hif-1αb of zebrafish in hypoxia/reoxygenation injury group were increased,Hif-1α protein levels were increased,brain infarct area was increased,cell apoptosis rate was increased,mitochondrial microstructure was seriously damaged,and GSH level was decreased,the protein levels of p-Akt/Akt,p-GSK-3β/GSK-3β and MAP2 were decreased,the fluorescence distribution and expression of MAP2 were decreased,and the frequency of optokinetic response was decreased;S Sevoflurane posttreatment and GSK-3β inhibitor TDZD-8 treatment can effectively improve the nerve injury of zebrafish under hypoxia/reoxygenation.Conclusion Akt/GSK-3β pathway is involved in the protective effect of sevoflurane posttreatment on zebrafish brain hypoxia/reoxygenation injury by affecting the expression of downstream tubulin MAP2. |
PDF Full Text Request