| Objective: By observing milalutide on the combined brain ischemic tissue of MiRNA-124,small glial cells/macrophages M1,M2 surface markers and their secretion of inflammatory factor tumor necrosis α(Tumor necrosis factor)-α,TNF-α),transforming growth factor-β(Transforming growth factor-β,TGF-β)to further explore the potential mechanisms of liraglutide neuroprotective effects.Method: Only 75 healthy male Sprague-Dawley rats were randomly divided into 5 groups,15 rats in each group.First,a permanent middle cerebral artery occlusion(MCAO)model was made using the thread occlusion method: cerebral ischemia(CI)group.The diabetic rat model was induced by intraperitoneal injection of streptozocin(STZ),and then the MCAO model was made by the suture method: diabetic cerebral ischemia(DCI)group,insulin group(Ins),liraglutide(Lir)Group,Liraglutide + MiRNA-124 antagonist group(Lir+ antagomir).miRNA-124 antagomir(0.2nom/μL,5 μL)was injected into the left side ventricle of the lir-antagomir group,and the remaining four groups of rats were injected with the same volume of physiological saline.One hour after the success of the MCAO molding,lir-antagomir and Lir groups were injected with liraglutide(700 μg/kg·d)in the abdominal cavity,Ins group was injected with corresponding amount of insulin every day;CI group and DCI group were injected with equal volume of normal saline every day.After MCAO molding 72 h,evaluation of neurological defects in rats,2,3,5-chlorinated diphenyl tetraazole(2,3,5-triphenyl tetrazolium,TTC)staining detection brain infarction volume,immunoprotein imprinting(western blot,WB)The content of TNF-α and TGF-β expression in brain tissue in ischemic regions is detected,and real-time fluorescence quantitative PCR(Real Time quantitative PCR,q RT-PCR)detects the expression levels of MiRNA-124,CD16,INOS,CD206,Arg-1 in brain tissue.Immuno-double fluorescence detects the co-labeling of M1 markers CD16/32 and M2 markers CD206 and Iba-1 in ischemic lateral brain tissue.Results:1.After STZ injection,the blood glucose levels of DCI group,Ins group,Lir group,Lir+antagomir group increased significantly(P<0.05,P<0.05,P<0.05);after drug intervention,Ins group,Lir group and The blood sugar level of Lir+ antagomir group decreased significantly(P<0.05,P<0.05,P<0.05).2.Neurological deficiency score:Compared to the CI group,the DCI group had a significantly higher neurological defect score(P<0.05),no significant change in the Ins group neurofunctional defect score compared to the DCI group(P>0.05),and Lir had a significantly lower neurological defect score(P<0.05)compared to the DCI group and lir-antagomir group,respectively.3.Volume of cerebral infarction: Different ranges of white infarction areas appeared in each group.Compared with the CI group,the volume of infarction in the DCI group increased significantly(P<0.05),while the volume of cerebral infarction in the Lir group was significantly reduced(P<0.05)compared to the DCI group,while there was no significant difference in the Ins group(P>0.05).Compared to the Lir group,the volume of cerebral infarction in the Lir-antagomir group increased significantly(P<0.05).4.Expression of MiRNA-124: Significant decrease in the expression of MiRNA-124 in the DCI group(P<0.05)compared to the CI group,significant increase in the expression of the MiRNA-124 expression in the Lir group(P<0.05)compared to the DCI group,and significant decrease in the expression of the MiRNA-124 expression in the Lir group(P<0.05)compared to the Lir group.5.The expression of the surface markers(CD16,i NOS)of type M1 microglia/macrophages and the secreted TNF-α: Compared with the CI group,the expression of CD16,i NOS and TNF-α in the DCI group increased significantly(P <0.05);Compared with the DCI group and the Lir+antagomir group,the expression levels of CD16,iNOS and TNF-α in the Lir group were significantly decreased(P<0.05).6.The expression of the surface markers(CD206,Arg-1)of type M2 microglia/macrophages and the secreted TGF-β : Compared with the CI group,the expression of CD206,Arg-1,TGF-β in the DCI group Significantly decreased(P<0.05);Compared with the DCI group and Lir+antagomir group,the expression levels of CD206,Arg-1 and TGF-β in the Lir group were significantly increased(P<0.05).7.M1 type microglia/macrophage expression: Compared with DCI group,the number of M1 type microglia/macrophage marker CD16/32/Iba-1 positive cells in Lir group was significantly reduced(P<0.05),Compared with the Lir group,the number of M1microglia/macrophage marker CD16/32/Iba-1 positive cells in the Lir+ antagomir group increased significantly(P<0.05).8.M2 type microglia/macrophage expression: Compared with the DCI group,the number of M2 type microglia/macrophages CD206/Iba-1 positive cells in the Lir group was significantly increased(P<0.05),which was similar to the Lir group Compared with that,the number of M2 type microglia/macrophages CD206/Iba-1 positive cells in the Lir+antagomir group was significantly reduced(P<0.05).Conclusion: Regulate the expression of MiRNA-124,inhibit the conversion of microglia/macrophages to the M1 phenotype,and promote the conversion of microglia/macrophages to the M2 phenotype,thereby reducing inflammation.It may be that liraglutide is associated with diabetes.One of the neuroprotective mechanisms of cerebral ischemic injury. |
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