心理应激致血浆外泌体miR-184-3p抑制血管内皮细胞增殖、迁移功能的作用分析

doi:10.11958/20212816

天津医药 ›› 2022, Vol. 50 ›› Issue (9): 927-931.doi: 10.11958/20212816

心理应激致血浆外泌体miR-184-3p抑制血管内皮细胞增殖、迁移功能的作用分析

苏晓雪¹^,²(), 黎静³, 赵威³, 刘林玲³, 刘素君³, 农腾川³, 谭机永¹^,²^,³^,^△()

1 广西医科大学转化医学研究中心（邮编530021）
2 长寿与老年相关疾病教育部重点实验室
3 广西医科大学基础医学院生理学教研室

收稿日期:2021-12-23 修回日期:2022-04-10 出版日期:2022-09-15 发布日期:2022-09-05
通讯作者: 谭机永 E-mail:527902103@qq.com;tanjiyong@yeah.net
作者简介:苏晓雪（1994）,女,硕士在读,主要从事心理应激与心血管疾病的发病机制方面研究。E-mail： 527902103@qq.com
基金资助:
广西自然科学基金项目(2020GXNSFAA159148)

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

SU Xiaoxue¹^,²(), LI Jing³, ZHAO Wei³, LIU Linling³, LIU Sujun³, NONG Tengchuan³, TAN Jiyong¹^,²^,³^,^△()

1 Centre for Translational Medicine, Guangxi Medical University, Nanning 530021, China
2 Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education
3 Teaching Section of Physiology, School of Basic Medicine, Guangxi Medical University

Received:2021-12-23 Revised:2022-04-10 Published:2022-09-15 Online:2022-09-05
Contact: TAN Jiyong E-mail:527902103@qq.com;tanjiyong@yeah.net

苏晓雪, 黎静, 赵威, 刘林玲, 刘素君, 农腾川, 谭机永. 心理应激致血浆外泌体miR-184-3p抑制血管内皮细胞增殖、迁移功能的作用分析[J]. 天津医药, 2022, 50(9): 927-931.

SU Xiaoxue, LI Jing, ZHAO Wei, LIU Linling, LIU Sujun, NONG Tengchuan, TAN Jiyong. Effects of chronic unpredictable mild stress induced plasma exosomal miR-184-3p on proliferation and migration of vascular endothelial cells[J]. Tianjin Medical Journal, 2022, 50(9): 927-931.

摘要/Abstract

摘要：

目的探讨心理应激源性血浆外泌体miR-184-3p在调控血管内皮细胞增殖、迁移功能中的作用。方法（1）将60只6~8周龄C57BL/6雄性小鼠随机分为对照组和心理应激组,每组30只。采用21 d慢性不可预知性心理应激刺激建模。通过行为学实验（旷场实验、高架十字迷宫实验和强迫游泳实验）评价建模效果。提取血浆外泌体进行电镜分析,qPCR检测血浆外泌体miR-184-3p表达水平。（2）体外培养小鼠肺微血管内皮细胞（MPVEC）,分别转染对照组小鼠血浆外泌体（C-EXO组）、心理应激组小鼠血浆外泌体（S-EXO组）、NC mimic组和miR-184-3p mimic组。划痕实验分析细胞迁移情况,EdU染色分析细胞增殖情况。结果（1）心理应激组小鼠探索行为较对照组显著降低,不动时间显著增加（P<0.01）。电镜分析验证血浆外泌体直径为30~200 nm的双层膜囊泡结构,qPCR证实心理应激组小鼠血浆外泌体中miR-184-3p的表达量显著增加（P<0.05）。（2）细胞实验结果显示S-EXO组细胞迁移和增殖能力较C-EXO组下降（P<0.01）;同时miR-184-3p mimic组细胞增殖和迁移能力较NC mimic组下降（P<0.05）。结论慢性心理应激源性血浆外泌体可以通过介导miR-184-3p抑制血管内皮细胞的增殖和迁移。

关键词: 应激,心理学, 细胞增殖, 细胞运动, 外泌体, miR-184-3p, 血管内皮细胞

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

中图分类号:

R541

图/表 8

表1 2组小鼠行为学检测指标的变化（n=11,s,$\bar{x}±s$）

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^＊＊

图1 超微透射电镜下血浆外泌体的形态结构（×15 000）

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

图2 血浆外泌体干预后血管内皮细胞划痕实验（×10）

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

表2 2组血管内皮细胞的迁移和增殖能力比较（n=3,%,$\bar{x}±s$）

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^＊＊

图3 血浆外泌体干预后血管内皮细胞EdU增殖染色情况（×20）

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

图4 miR-184-3p对血管内皮细胞划痕实验的影响（×20）

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

表3 2组血管内皮细胞的迁移和增殖能力比较（n=3,%,$\bar{x}±s$）

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^＊

图5 miR-184-3p对血管内皮细胞EdU增殖染色的影响（×20）

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

参考文献 18

[1]	陈帅, 陆登成, 何丽敏, 等. 心理应激对心血管功能的影响[J]. 中国继续医学教育, 2019, 11(36):158-160.
	CHEN S, LU D C, HE L M, et al. The effect of psychological stress on cardiovascular function[J]. CCME, 2019, 11(36):158-160. doi: 10.3969/j.issn.1674-9308.2019.3.061.
[2]	TODA N, NAKANISHI-TODA M. How mental stress affects endothelial function[J]. Pflugers Arch, 2011, 462(6):779-794. doi: 10.1007/s00424-011-1022-6.
[3]	黎静, 高金洪, 莫振华, 等. 海带多糖对心理应激大鼠血管舒张功能的保护作用[J]. 中草药, 2012, 43(4):748-751.
	LI J, GAO J H, MO Z H, et al. Protection of laminaria polysaccharide on vasodilation function of rats induced by psychological stress[J]. Chinese Traditional and Herbal Drugs, 2012, 43(4):748-751.
[4]	FERNÁNDEZ-HERNANDO C, SUÁREZ Y. MicroRNAs in endothelial cell homeostasis and vascular disease[J]. Curr Opin Hematol, 2018, 25(3):227-236. doi: 10.1097/MOH.0000000000000424.
[5]	FANG K, XU J X, CHEN X X, et al. Differential serum exosome microRNA profile in a stress-induced depression rat model[J]. J Affect Disord, 2020, 274:144-158. doi: 10.1016/j.jad.2020.05.017.
[6]	胡力旬, 田锋. 心理应激在心血管病发生中的病理生理机制研究[J]. 中西医结合心血管病电子杂志, 2016, 4(20):2-3.
	HU L X, TIAN F. Psychological stress in heart vascular disease occurred in the pathophysiological mechanism research[J]. Cardiovascular Disease Journal of Integrated Traditional Chinese and Western Medicine, 2016, 4(20):2-3. doi: 10.3969/j.issn.2095-6681.2016. 20.002.
[7]	LI N, WANG Q, WANG Y, et al. Fecal microbiota transplantation from chronic unpredictable mild stress mice donors affects anxiety-like and depression-like behavior in recipient mice via the gut microbiota-inflammation-brain axis[J]. Stress, 2019, 22(5):592-602. doi: 10.1080/10253890.2019.1617267.
[8]	王彬, 齐县伟, 张宪亮. MiR-193a-3p通过调控S100A4表达对oxLDL诱导的血管内皮细胞损伤影响[J]. 中国循证心血管医学杂志, 2021, 13(6):742-746,751.
	WANG B, QI XW, ZHANG X L. Influence of miR-193a-3p on oxidized low density lipoprotein-induced vascular endothelial cell injury through regulating S100A4 expression[J]. Chinese Journal of Evidence-Bases Cardiovascular Medicine, 2021, 13(6):742-746,751. doi: 10.3969/j.issn.1674-4055.2021.06.25.
[9]	葛丽, 许红峰. miR-181b在动脉粥样硬化血管中的表达及对H₂O₂诱导的血管内皮细胞增殖与凋亡的影响[J]. 解剖科学进展, 2021, 27(1):9-13.
	GE L, XU H F. Expression of miR-181b in atherosclerotic vessels and its effect on proliferation and apoptosis of vascular endothelial cells induced by H₂O₂[J]. Progress of Anatomical Sciences, 2021, 27(1):9-13. doi: 10.16695/j.cnki.1006-2947.2021.01.003.
[10]	张煜, 张洪伟, 杨鹏, 等. 神经纤毛蛋白-1调控血管内皮细胞增殖迁移的功能作用和潜在机制研究[J]. 中国心血管病研究, 2021, 19(6):570-576.
	ZHANG Y, ZHANG H W, YANG P, et al. The role and potential mechanisms of neuropilin-1 in regulating the proliferation and migration of vascular endothelial cells[J]. Chinese Journal of Cardiovascular Research, 2021, 19(6):570-576. doi: 10.3969/j.issn.1672-5301.2021.06.017.
[11]	WORTZEL I, DROR S, KENIFIC C M, et al. Exosome-mediated metastasis: Communication from a distance[J]. Dev Cell, 2019, 49(3):347-360. doi: 10.1016/j.devcel.2019.04.011.
[12]	武庆娟, 陈恒文, 高健, 等. 外泌体miRNA在心血管疾病中的研究进展[J]. 中国病理生理杂志, 2020, 36(2):371-377.
	WU Q J, CHEN H W, GAO J, et al. Exosome-derived miRNAs in cardiovascular diseases[J]. Chinese Journal of Pathophysiology, 2020, 36(2):371-377. doi: 10.3969/j.issn.1000-4718.2020.02.027.
[13]	周正阳. 外泌体运载的miR-155靶向血管内皮细胞FOXO3促进胃癌血管生成[D]. 天津: 天津医科大学, 2020.
	ZHOU Z Y. Exosomes carrying miR-155 target FOXO3 of endothelial cells and promote angiogenesis in gastric cancer[D]. Tianjin: Tianjin Medical University, 2020.
[14]	JING R, ZHONG Q Q, LONG T Y, et al. Downregulated miRNA-26a-5p induces the apoptosis of endothelial cells in coronary heart disease by inhibiting PI3K/AKT pathway[J]. Eur Rev Med Pharmacol Sci, 2019, 23(11):4940-4947. doi: 10.26355/eurrev_201906_18084.
[15]	张晓蕾, 许国莹, 王士珍, 等. miR-186-5p对冠心病大鼠血管内皮细胞损伤及FGF2/FGFR1信号通路的影响[J]. 天津医药, 2021, 49(11):1169-1174.
	ZHANG X L, XU G Y, WANG S Z, et al. Effects of miR-186-5p on vascular endothelial cell injury and FGF2/FGFR1 signaling pathway in rat model of coronary heart disease[J]. Tianjin Med J, 2021, 49(11):1169-1174. doi: 10.11958/20211008.
[16]	DILIP S, KARMYODH S, PRAGNYA D, et al. miR-184 mediates hyperoxia-induced injury by targeting cell death and angiogenesis signalling pathways in the developing lung[J]. Eur Respir J, 2021, 58(3):1901789. doi: 10.1183/ 13993003.01789-2019.
[17]	PARK J K, PENG H, YANG W, et al. miR-184 exhibits angiostatic properties via regulation of Akt and VEGF signaling pathways[J]. FASEB J, 2017, 31(1):256-265. doi: 10.1096/fj.201600746R.
[18]	LU T X, ROTHENBERG M E. MicroRNA[J]. J Allergy Clin Immunol, 2018, 141(4):1202-1207. doi: 10.1016/j.jaci.2017.08.034.

心理应激致血浆外泌体miR-184-3p抑制血管内皮细胞增殖、迁移功能的作用分析

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

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 8

参考文献 18

相关文章 15

编辑推荐

Metrics

本文评价

[1]	张晋玮, 王燕, 王通. miR-107对口腔鳞癌细胞系CAL27增殖、侵袭及迁移的影响[J]. 天津医药, 2024, 52(9): 897-899.
[2]	马佳佳, 张亚苹, 杨斌, 赵美琪, 蒋璐, 黄小玉, 范路畅, 王凤梅. ATOX1通过JAK2/STAT3通路促进肝癌细胞生物学行为的机制探讨[J]. 天津医药, 2024, 52(9): 907-912.
[3]	周丽芸, 王岩, 董本超, 杨培川, 申佳慧, 马剑雄, 马信龙. 干细胞机械敏感性对抗骨质疏松症研究进展[J]. 天津医药, 2024, 52(8): 877-881.
[4]	杨睿, 魏琼, 孙逸坤, 赵梦竹, 程序, 刘梦华, 张冬梅. 缺氧H9c2来源外泌体对HUVEC增殖、迁移和成管能力的影响[J]. 天津医药, 2024, 52(7): 714-719.
[5]	刘丹阳, 李永涛, 张海燕, 李林, 刘洋, 沈雷. 乳腺癌细胞条件培养基对骨髓间充质干细胞生物学行为的影响[J]. 天津医药, 2024, 52(5): 454-458.
[6]	王鑫瑶, 杨惠, 李冰冰. 间充质干细胞在子宫内膜异位症中的研究进展[J]. 天津医药, 2024, 52(2): 215-219.
[7]	林瑶, 刘从娜, 王世霞, 张志勇. 金合欢素调节HMGB1/TLR4信号通路对脂多糖诱导牙髓细胞凋亡的影响[J]. 天津医药, 2024, 52(12): 1238-1243.
[8]	张贵婷, 何超. oxLDL/β2GPⅠ/aβ2GPⅠ复合物通过TLR4/MyD88/NF-κB途径促进血管内皮细胞血管生成[J]. 天津医药, 2024, 52(11): 1131-1136.
[9]	郝凯凯, 王晓敏, 刘峥, 刘东洋, 李静. 藁本内酯调节RhoA/ROCK信号通路对食管癌细胞生物学行为的影响[J]. 天津医药, 2024, 52(11): 1164-1170.
[10]	孙创新, 李刚. NID1在肾透明细胞癌血管生成中的作用研究[J]. 天津医药, 2024, 52(10): 1009-1013.
[11]	姚和, 林艳丽, 崔雨萌, 阎新龙. 外泌体环状RNA在胃癌中作用机制的研究进展[J]. 天津医药, 2024, 52(10): 1110-1115.
[12]	徐桂颖, 李玉, 李雪, 刘怡萌, 陈怀永. 气道类器官研究哮喘中Lkb1调控上皮再生的机制[J]. 天津医药, 2024, 52(1): 11-15.
[13]	张琳琳, 赵唐明, 黄婵, 李善文, 甘卫华. 新型多肽AMPP2在TGF-β1诱导的系膜细胞增殖中的作用及其机制[J]. 天津医药, 2024, 52(1): 50-55.
[14]	邓乾葆, 张忠霞, 王茹, 许琳, 罗书. 基于外泌体多miRNA表达水平构建的子痫前期风险预测模型效能分析[J]. 天津医药, 2024, 52(1): 91-96.
[15]	侯永波, 余骏马, 朱海娟. 外泌体在心肌梗死治疗中的研究新进展[J]. 天津医药, 2023, 51(9): 1016-1019.