Определение концентрации холестерина и триглицеридов в экстраклеточных везикулах сыворотки крови с помощью коммерческих наборов

Exosomes and microvesicles, collectively referred to as small extracellular vesicles (sEV) are vesicles with an average size of about 100-150 nm. Currently, the role of sEV in various aspects of signaling in the body is being actively investigated; in addition, sEV can often serve as markers of various pathologies. The active study of the sEV composition is continuing. In this study we have demonstrated that in sEV it is possible to determine cholesterol and triglycerides concentration by using commercial kits designed for serum. The technique was tested on sEV from the blood of patients diagnosed with depression and on healthy volunteers. No differences were found in the concentration of cholesterol and triglycerides in mEV from the blood serum of depressed patients and the control group. The concentration of cholesterol and triglycerides in the samples is several times higher than the sensitivity threshold of the methods set by the manufacturer of the kits.Экзосомы и микровезикулы, совместно называемые малые экстраклеточные везикулы (мЭВ), представляют собой везикулы со средним размером около 100-150 нм. В настоящее время активно исследуется роль мЭВ в самых разных аспектах сигналинга в организме, кроме того, часто мЭВ могут служить маркерами разных патологий. Продолжается активное изучение состава мЭВ. В данной работе мы показали, что в мЭВ можно определить концентрацию холестерина и триглицеридов с помощью коммерческих наборов, предназначенных для сыворотки крови. Методика была опробована на мЭВ из крови пациентов с диагнозом депрессия и на здоровых добровольцах. Различий в концентрации холестерина и триглицеридов в мЭВ из сыворотки крови пациентов с депрессией и контрольной группы найдено не было. Концентрация холестерина и триглицеридов в образцах в несколько раз превосходит порог чувствительности методов, установленный производителем наборов

Чувствительность экстраклеточных везикул из сыворотки крови человека к различным детергентам

Blood exosomes and microvesicles, collectively known as small extracellular vesicles (sEV), are vesicles about 100-150 nm in size. Small EV are involved in various aspects of signaling in the body; in addition, they can serve as markers of various pathologies. For biochemical studies, vesicle solubilization is often required. We tested the ability of various detergents to dissolve membranes of the sEV. Small EV were isolated from the blood serum of healthy volunteers by gel filtration on Sepharose CL-2B and tried to solubilize them using the anionic detergent DOC (sodium deoxycholate), non-ionic detergent Brij 35 (polyoxyethyleneglycol dodecyl ether), zwitterionic detergent CHAPS (3 - [(3-chloramidopropyl) dimethylammonio] -1-propanesulfonate), and cationic detergent CTAB (cetyl trimethylammonium bromide). The concentration of sEV in the solution was determined by dynamic light scattering. We find DOC is the most effective for sEV solubilization.Экзосомы и микровезикулы, совместно называемые малые экстраклеточные везикулы (мЭВ), представляют собой везикулы размером около 100-150 нм. Малые ЭВ принимают участие в самых разных аспектах сигналинга в организме; кроме того, они могут служить маркерами разных патологий. Для биохимических исследований зачастую требуется солюбилизация везикул. Мы проверили способность различных детергентов растворять мембраны мЭВ. Малые ЭВ выделяли из сыворотки крови здоровых добровольцев с помощью гель-фильтрации на Sepharose CL-2B и пытались их солюбилизировать с помощью анионного детергента DOC (дезоксихолат натрия), неионного детергента Brij 35 (додециловый эфир полиэтиленгликоля), цвиттерионного детергента CHAPS (3-[(3-хлорамидопропил) диметиламмонио]-1-пропансульфонат), катионного детергента CTAB (цетил триметиламмония бромид). Концентрацию мЭВ в растворе определяли с помощью динамического светорассеяния. Самым эффективным детергентом для солюбилизации мЭВ из сыворотки крови оказался DOC

The Missing Link: How Exosomes and miRNAs can Help in Bridging Psychiatry and Molecular Biology in the Context of Depression, Bipolar Disorder and Schizophrenia

MicroRNAs (miRNAs) only recently have been recognized as promising molecules for both fundamental and clinical neuroscience. We provide a literature review of miRNA biomarker studies in three most prominent psychiatric disorders (depression, bipolar disorder and schizophrenia) with the particular focus on depression due to its social and healthcare importance. Our search resulted in 191 unique miRNAs across 35 human studies measuring miRNA levels in blood, serum or plasma. 30 miRNAs replicated in more than one study. Most miRNAs targeted neuroplasticity and neurodevelopment pathways. Various limitations do not allow us to make firm conclusions on clinical potential of studied miRNAs. Based on our results we discuss the rationale for future research investigations of exosomal mechanisms to overcome methodological caveats both in studying etiology and pathogenesis, and providing an objective back-up for clinical decisions. © 2019, Springer Science+Business Media, LLC, part of Springer Nature

Elevated Levels of Serum Exosomes in Patients with Major Depressive Disorder

Exosomes are small (30-100 nm) vesicles of the endocytic membrane that may be found in the endosome system of multivesicular organs and are released after fusion of multivesicular bodies with the plasma membrane. Exosomes function as carriers for specific loads and are stable in biological fluids; hence, exosomes are considered as potential biomarkers of different diseases. The degree of involvement and functional significance of exosomes in pathogenesis of depression have been poorly studied. The development of diagnostic methods for depressive disorders based on the analysis of circulating exosomes is very promising, despite the methodological complications related to their isolation and identification. The purpose of this study was to isolate exosomes from the blood serum of patients with major depressive disorder (MDD) and compare them with healthy volunteers using different methods of evaluation. The concentration of serum exosomes assessed using the methods of dynamic light scattering, nanoparticle tracking analysis, and the enzyme-linked immunoassay was statistically elevated in patients with MDD compared to healthy volunteers. The possible connection between the increased level of serum exosomes with the cell-mediated immune activation that is seen in depression is discussed