Effects of chronic unpredictable mild stress induced plasma exosomal miR-184-3p on proliferation and migration of vascular endothelial cells

doi:10.11958/20212816

Abstract

Abstract:

Objective To investigate the role of psychological stressor plasma exosome miR-184-3p in regulating the proliferation and migration of vascular endothelial cells. Methods (1) Sixty male C57BL/6 mice aged 6 to 8 weeks were randomly divided into the control group and the psychological stress group, with 30 mice in each group. The model of 21 d chronic unpredictable psychological stress was used. Behavioral experiments (open field experiment, elevated cross maze experiment and forced swimming experiment) were used to evaluate the modeling effect. Plasma exosomes were extracted and analyzed by electron microscopy. The expression level of plasma exosome miR-184-3p was detected by qPCR. (2) Mouse lung microvascular endothelial cells (MPVEC) were cultured in vitro and transfected with plasma exosome of control mice (C-EXO group), plasma exosome of psychological stress group (S-EXO group), NC mimic and miR-184-3p mimic, respectively. Cell migration was analyzed by scratch test, and cell proliferation was analyzed by EdU staining. Results (1) Compared with the control group, the exploration behavior of mice in the psychological stress group was significantly decreased, and the immobility time was significantly increased (P<0.01). The structure of bilayer membrane vesicles with plasma exosomes diameter of 30-200 nm was verified by electron microscope analysis. qPCR confirmed that the expression level of miR-184-3p in plasma exosomes of mice in the psychological stress group was significantly increased (P<0.05). (2)The results of cell experiment showed that the migration and proliferation of the S-EXO group were lower than that of the C-EXO group (P<0.01). Meanwhile, the proliferation and migration of the miR-184-3P mimic group were decreased compared with the NC mimic group (P<0.05). Conclusion Chronic psychological stressor plasma exosomes can inhibit the proliferation and migration of vascular endothelial cells by mediating miR-184-3p.

Key words: stress,psychological, cell proliferation, cell movement, exosome, miR-184-3p, endothelial cells

CLC Number:

R541

Figures/Tables 8

Tab.1 Changes of behavioral test indexes in two groups of mice

组别	旷场实验（中央区域停留时间）	高架十字迷宫实验（开放臂停留时间）	游泳实验（漂浮不动时间）
对照组	12.18±5.41	114.50±12.51	241.70±18.70
心理应激组	6.36±1.32	104.50± 8.94	313.50±8.28
t	3.454^＊＊	2.176^＊	11.650^＊＊

Fig.1 Morphology and structure of plasma exosomes under ultratransmission electron microscopy (×15 000)

Fig.2 Scratch test of vascular endothelial cells after plasma exosome intervention (×10)

Tab.2 Comparison of migration and proliferation of vascular endothelial cells between two groups

组别	划痕迁移率	EdU增殖率
C-EXO组	57.53±8.66	69.67±1.53
S-EXO组	4.31±2.64	53.33±5.51
t	10.180^＊＊	4.950^＊＊

Fig.3 EdU staining of vascular endothelial cells after plasma exosome intervention (×20)

Fig.4 Effects of miR-184-3p on the scratch experiment of vascular endothelial cells (×20)

Tab.3 Comparison of migration and proliferation of vascular endothelial cells between two groups

组别	划痕迁移率	EdU增殖率
NC mimic组	58.30±6.33	55.80±1.91
miR-184-3p mimic组	42.97±1.86	45.60±4.62
t	4.025^＊	3.534^＊

Fig.5 Effects of miR-184-3p on proliferation and EdU staining of endothelial cells（×20）

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