The Relationship Between The Expression Of Plasma MiR-202-5p And The Condition And Prognosis Of Patients With Aneurysmal Subarachnoid Hemorrhage

Objective: The expression level of plasma exosome miR-202-5p in patients with aneurysmal subarachnoid hemorrhage(aSAH)was detected,and its relationship with the condition and prognosis of patients with aSAH was discussed.Method: 1.39 aSAH patients admitted to the department of neurology and department of neurosurgery,the first affiliated hospital of guangxi medical university from June 2017 to June 2018 were selected as the research objects,and 20 healthy people matched in gender and age were used as the normal control group.According to hunt-hess grading method,the subjects were divided into mild disease group and severe disease group,and mRS scores 3 months after discharge were divided into good prognosis group and poor prognosis group.2.Peripheral venous blood samples were collected,centrifuged and extracted to separate plasma exosomes.Exosomes were identified by transmission electron microscopy and plasma exosome RNA was extracted.On the basis of another study in our group,12 patients with intracranial aneurysms(4 cases of unruptured,mild and severe)and 4 healthy patients were analyzed byhigh-throughput sequencing,and the miRNA expression profile of plasma exosomes was obtained,and miR-202-5p with significant difference in expression was screened for PCR verification.The relative expression levels of miR-202-5p in plasma samples of the normal group and the case group were determined by real-time quantitative PCR.3.SPSS22.0 software was used for data statistical analysis.T-test was used for normal distribution of measurement data,non-normal distribution of measurement data was used for rank sum test,and chi-square test was used for categorical variable data,P < 0.05 was statistically significant.Plasma exosome miRNA levels were compared between different groups.ROC curves were drawn based on experimental data to compare the diagnostic value of plasma exosome miRNA in patients with severe aSAH and poor prognosis.Results:1.Clinical data analysis showed that severe patients had higher WFNS score,Fisher score and mRS score,larger aneurysm diameter(?5mm),higher proportion of history of consciousness disorder and pneumonia than mild patients;Compared with patients with good prognosis,patients with poor prognosis had higher WFNS score and Fisher score,and had higher proportion of history of disturbance of consciousness and pneumonia.2.Using transmission electron microscope to observe the exosome morphology,it can be seen that exosomes are round or quasi-round,with double-layer lipid molecular structure,with uneven size and distribution,and the diameter is between 20 and 200 nm,which is in line with the normal morphological characteristics of exosomes.3.High-throughput results showed that there were 6 mirnas that met the conditions between the normal group and the non-ruptured group.Between the normal group and the ruptured group,there were 10 mirnas that met the conditions.Between the unruptured group and the ruptured group,there were 23 mirnas that met the conditions.Seven mirnas qualified between the severe and mild disease groups.The analysis and intersection of differentially expressed mirnas between each group indicated significant differences in the expression of miR-202-5p between different groups.4.Through bioinformatics analysis of miR-202-5p,the biological process,cell components and molecular functions targeted by miR-202-5p all indicate that miR-202-5p may be related to the rupture process of intracranial aneurysms.The enrichment pathway of KEGG suggested that miR-202-5p was involved in the physiological and pathological processes of various diseases.5.PCR verification of miR-202-5p with expanded samples showed that,compared with the normal group,the expression level of plasma exosome miR-202-5p was significantly increased in the aSAH group(P<0.05).Compared with the mild group,the relative expression level of miR-202-5p was up-regulated in the severe group(P<0.05).Compared with the group with good prognosis,the expression level of miR-202-5p was up-regulated in the group with poor prognosis(P<0.05).ROC curve analysis showed that the area under the curve predicted by plasma exosome miR-202-5p level in severe aSAH patients was 0.758(95%CI,0.607-0.908).The area under the curve predicted poor prognosis of aSAH patients was 0.666(95%CI,0.494-0.837).Conclusion:Plasma exosome miR-202-5p is highly expressed in aSAH patients,which may be an indicator of the severity and prognosis of patients with aSAH.