Expression And Significance Of Plasma Exosomal Mir-125b-3p In Patients With Cerebral Infarction

Objective:To detect the expression of exosomal miR-125b-2-3p in plasma of patients with acute cerebral infarction,subacute stage cerebral infarction and recovery stage cerebral infarction,and compared with 25 age and sex matched healthy controls,to investigate the the potential value of plasma exosomal miR-125b-2-3p as an early biomarker of cerebral infarction.Method:We enrolled 90 patients with cerebral infarction and 25 healthy controls.We divided patients into three groups,acute cerebral infarction(1~7d,30),subacute stage cerebral infarction(8~14d,30)and recovery stage cerebral infarction(15~180d,30).The patients were selected for NIHSS score at admission,according to the results were divided into three levels of light weight,Mild <5,moderate: 5~20 points,severe > 20 points,of which mild in 52 cases,moderate in 34 cases,severe in 4 cases.According to the TOAST classification criteria,the patients with cerebral infarction were divided into five groups:(1)LAA:62 cases;(2)SAA:15 cases;(3)CEI:5 cases;(4)SUE:4 cases;(5)SOE:4cases.Exosomes were isolated from the plasma of the patients and control group by exoRNeasy kit.We observed morphnology of exosomes by transmission electron microscope and measure the particle size measurement by ZETASIZER Nano series-Nano-ZS,teste the membrane marker of exosomes in vitro by flow cytometry.Screened the ExoCarta database and combined with literature reports,we selected exosomal miR-125b-2-3p to research,the expression of exosomal miR-125b-2-3p in plasma of patients with cerebral infarction in different course of disease was confirmed by qRT-PCR method,and to analyze the relationship between neurological deficit and TOAST classification.Rrsults:1?Exosomes was round or oval by transmission electron microscope,with a lipid bilayer molecular structure.The diameter were between 20~200nm,the particle size was 186.9nm,the average diameter were 128.2nm,and the particle size distribution of 20nm-200 nm was 65.1%,which was consistent with the size distribution of the exosomes of the particle diameter.And the surface marker protein CD63 and CD81 were detected positive,defining as exosomes.2?Compared with the healthy control group,the expression of exosomal miR-125b-2-3p in the patients with acute and subacute stage and recovery stage of cerebral infarction group was up-regulated(P<0.05),There was no significant difference between the three groups in the cerebral infarction group(P>0.05).3?According to the NIHSS score,the patients with cerebral infarctionwere divided into three groups: mild,moderate and severe.The difference of the expression of exosomal miR-125b-2-3p in the patients with mild to moderate severe cerebral infarction was not statistically significant(P>0.05),the expression of exosomal miR-125b-2-3p was not related to the severity of neurological deficit.4?The cerebral infarction group was divided into five groups(TOAST type),the expression of exosomal miR-125b-2-3p in the LAA group was higher than that in the other groups,but there was no significant difference between the two groups(P> 0.05).5?The cause of disease of cerebral infarction group(TOAST)each subgroup comparison,the expression of exosomal mi R-125b-2-3p in the five groups was up-regulated,but there was no significant difference between the two groups(P> 0.05).6?In the acute and subacute stage and recovery stage of cerebral infarction patients,the expression of exosomal miR-125b-2-3p in the ROC curve area under the AUC were 0.790,0.923,0.793,suggesting that exoskeletal miR-125b-2-3p had the diagnostic value in all stages of cerebral infarction.The area under the ROC curve of the exogenous pathogenic miR-125b-2-3p in subacute cerebral infarction was the largest,AUC = 0.923,The source of miR-125b-2-3p is more valuable in the diagnosis of subacute cerebral infarction.Conclusion:1?We successfully isolated and identified the exosomes from plasma.2?The expression of exosomal miR-125b-2-3p in the plasma of patients with cerebral infarction was significantly up-regulated,which may be a biomarker for the diagnosis of cerebral infarction.