Correlation Study Of Peripheral Blood Exosomal MicroRNA As Moyamoya Disease Diagnostic Markers

Objective: To search for differential miRNAs transported by exosomes of MMD blood origin,to identify the sensitivity and specificity of differential target miRNAs in MMD diagnosis,to specify 1-2 target miRNAs as markers for MMD diagnosis,and to assess the clinical value of blood miRNAs of exosomal origin in MMD diagnosis and treatment,help for MMD diagnostics.Methods: The experimental group was selected from 30 patients hospitalized in the affiliated hospital of Jining Medical College,Shandong Province,China with a diagnosis of MMD in neurosurgery from September 2019 to July 2022,and the control group was selected from 20 healthy individuals who had health checkups at the physical examination center from September 2019 to July 2022,and general information such as age,sex,height,weight history of alcohol consumption and smoking history,as well as chief complaints,CT perfusion imaging,DSA imaging were collected from the patients Clinical information.Fasting venous blood（10 ml）was collected from patients in the morning,centrifuged and stored in a-80°C refrigerator.Exosomes were extracted from the blood of MMD patients and healthy people using Exquick-related technology.All RNAs carried by the extracted exosomes were sequenced and analyzed using a sequencing platform to find differential miRNAs.Select the target miRNA for PCR verification and use SPSS software to draw ROC curves,analyze specificity and sensitivity.Results:1.Under transmission electron microscopy,the exosomass exhibits a cup or tea tray,and the size of the vesicle is distributed between 40-150 nm,the specific marker protein CD9 of the outer body,the TSG101 is obviously enriched,and the negative marker Calnexin is not expressed.2.The concentration of exosome from MMD patient’s plasma（7.02E+10-8.62E+10Particles/m L）is higher than that from healthy control（6.52E+10-8.69E+10Particles/m L）.3.High-throughput sequencing display: There is a significant difference between micro RNA from MMD and healthy control,there are196 expressions,131 expressions are lowered.4.Differential Micro RNA target genes are mainly involved in intercellular connections,proteins,enzymes,and cationic binding,cell transformation,and primary metabolism,and associated with neurotrophic signal transduction and MAPK signaling pathways.5.hsa-miR-151a-3p、hsa-miR-200c-3p+hsa-miR-128-3p、hsa-miR-200c-3p+hsa-miR-483-5p 、 hsa-miR-99b-5p+hsa-miR-483-5p shows strong specificity and sensitivity in ROC curve analysis,the outer area of the curve is 1,which can be used as an indicator of early screening of MMD.Conclusion: This study found that there was a significant difference in miRNA of peripheral blood Exosome source between MMD and health control groups,which had potential value for MMD diagnosis.Screening the height-specific and sensitive new indicators（hsa-miR-151a-3p）and Combination indicators（hsa-miR-200c-3p+hsa-miR-128-3p 、hsa-miR-200c-3p+hsa-miR-483-5p、hsa-miR-99b-5p+hsa-miR-483-5p）for the early screening of MMD.