Correlation Of MiR-126 Expression With Traditional Chinese Medicine Syndrome And The Change And Effect Of MiR-126 After Acute Ischemic Stroke

Chapter I Study on the Relationship between miR-126 and SPREDI/VEGFA Expression Levels and Traditional Chinese Medicine Classification in Patients with Acute Ischemic StrokeObjective:This study clarified how serum miR-126 and SPREDI/VEGFA were correlated with different Traditional Chinese Medicine syndromes of acute ischemic stroke,and analyzed the possible mechanism of different Traditional Chinese Medicine syndrome types of acute ischemic stroke,so as to provide an early objective basis for the diagnosis,classification and prognosis of acute ischemic stroke in Chinese and western medicine.Methods:A method was used to collect clinically acceptable acute ischemic stroke patients(108 cases)as the experimental group,and non-stroke healthy patients(30 cases)were used as the control group.After Traditional Chinese Medicine syndrome differentiation,the patients were tested for serum miR-126,SPRED1 and VEGFA indicators,and completed the NISHH scale and Barthel index score,which was to explore how serum miR-126 and SPRED1/VEGFA were correlated with different Traditional Chinese Medicine syndromes of acute ischemic stroke.Results:1.The level of miR-126 in serum of patients with acute ischemic stroke was significantly lower than that in the control group,and the decrease in wind-phlegm-stasis syndrome was significantly different from the other three syndrome types(p<0.05).2.The expression levels of serum SPRED1 and VEGFA in the experimental group were significantly higher than those in the control group.The difference was that in the four syndrome types,the serum SPRED1 in Phlegm-heat accumulation type was significantly different from the other syndromes,while serum VEGFA was significantly different in wind-phlegm-stasis type(p<0.05)Conclusion:Serum miR-126 and VEGFA were the most significant expressions in wind-phlegm-stasis type in patients with acute ischemic stroke.Therefore,serum miR-126 combined with VEGFA could be used as an objective indicator of wind-phlegm-stasis type in acute ischemic stroke.Chapter II Dynamic Changes in miR-126 Expression in the Hippocampus and Penumbra Following Experimental Transient Global and Focal Cerebral Ischemia-ReperfusionObjective:To investigate the change of miR-126 with acute ischemia-reperfusion time in different brain tissues after global and focal cerebral ischemia and clarify the mechanism of miR-126 through SPRED1/NEGFA/Raf-1 signaling pathway to regulate neuron injury after acute ischemic stroke.Methods:ICR mice were chosen to establish the middle cerebral artery occlusion model by MCAO and Mongolian gerbils were chosen to establish the global cerebral ischemia model by ligation of bilateral common carotid arteries.Cerebral infarction volume was assessed by TTC staining.Longa scoring was used to observe neurological impairment.The changes of GFAP,Iba-1 and NeuN immunoreactivity in hippocampal CA1 region was observed by Immunohistochemical staining.Cresyl staining was used to evaluate Neuronal damage.PC 12 cells were chosen to establish(oxygen and glucose deprivation and reperfusion)OGD/R model,cell activity was detected by CCK-8 method,and siRNA low expression plasmid was used to constructe the low expression model of miR-126,SPRED1 and VEGFA in vitro.The expression of miR-126-3p and miR-126-5p at the RNA level after ischemia-reperfusion in vitro and in vivo were determined by RT-PCR.The changes of SPREDINEGFA/Raf-1 pathway proteins were observed by Western blotting.Results:1.Decreases in the levels of miR-126-3p and miR-126-5p expression in mice and gerbils after acute ischemic stroke,consistent with the results after OGD/R in PC12 cells.2.Glial cells were activated as neuronal damage gradually increased after acute ischemic stroke.3 Inhibition of miR-126-3p exacerbated the OGD/R-induced damage of cell death and reduced cell viability.4.After miR-126-3p inhibition,the levels of SPRED1 and VEGFA expression were increased,and p-Raf-1 expression was decreased after OGD/R.Moreover,based on the intervention of miR-126-3p inhibition,we found that the expression of fp-Raf-1 was significantly increased after the intervention of siSPRED1,while it was not statistically significant after intervention of siVEGFAConclusion:Dynamic changes in miR-126 expression were related to neuronal damage after acute ischemia-reperfusion in both focal and global transient cerebral ischemia models.In addition,the experiment showed that the reduction of miR-126 after global and focal cerebral ischemia exacerbated neuronal death,which was intimately related to increasing the SPRED1 activation and inhibiting the Raf-1 expression.This study showed that miR-126 may be a potential target for pharmacogenetic treatment in patients with acute ischemic stroke in the future.