Spinal cord injury alters microRNA and CD81+ exosome levels in plasma extracellular nanoparticles with neuroinflammatory potential.
Spinal cord injury alters microRNA and CD81+ exosome levels in plasma extracellular nanoparticles with neuroinflammatory potential.
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DOI：
10.1016/j.bbi.2020.12.007
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Wu J
中科院分区：
文献类型：
--
作者：
Khan NZ;Cao T;He J;Ritzel RM;Li Y;Henry RJ;Colson C;Stoica BA;Faden AI;Wu J
细胞外囊泡（EV）在机制上与神经退行性疾病（包括中枢神经系统损伤）的病理生物学有关。然而，迄今为止，EV 在脊髓损伤（SCI）中的作用受到的关注还很有限。此外，与 EV 隔离和表征方法相关的技术限制可能会导致误导或矛盾的发现。在这里，我们使用补充测量技术检查了小鼠 SCI 后多个时间点（1d、3d、7d、14d）血浆 EV 的变化。如纳米颗粒追踪分析 (NTA) 所示，通过超速离心 (UC) 分离的血浆 EV 在损伤后第 1 天减少，并且与血浆细胞外纳米颗粒总数的总体减少相平行。 UC 来源的血浆 EV 的蛋白质印迹 (WB) 分析显示，在损伤后 1 天到 7 天之间，四跨膜蛋白外泌体标记物 CD81 的表达增加。为了证实这些发现，我们对使用 ExoView® 直接从血浆捕获的单个四跨膜 EV 进行了干涉和荧光成像。与 WB 一致，我们观察到受伤后 1 天血浆 CD81+ EV 计数和货物显着增加。根据干涉测量法，这些四跨膜蛋白 EV 中的大多数小于 50 nm，并且通过基于流式细胞术的检测无法充分解析。在损伤部位，EV 生物发生蛋白的表达增强，在 WB 直接从脊髓组织分离的 EV 中也检测到了这种蛋白。损伤部位多种细胞类型中四跨膜蛋白 CD9 和 CD63 的表面表达增加；然而，流式细胞术证明，星形胶质细胞 CD81 表达独特下降。 UC分离的血浆EV microRNA货物在损伤后1天也发生显着改变，其变化与星形胶质细胞在炎症刺激后释放的EV中报道的变化相似。当注射到侧脑室时，来自 SCI 小鼠的血浆 EV 增加了大脑皮层中促炎和抗炎基因以及反应性星形胶质细胞基因的表达。这些研究首次提供了 SCI 后血浆 EV 动力学的详细表征，并表明血浆 EV 可能与创伤后脑炎症有关。
Extracellular vesicles (EVs) have been implicated mechanistically in the pathobiology of neurodegenerative disorders, including central nervous system injury. However, the role of EVs in spinal cord injury (SCI) has received limited attention to date. Moreover, technical limitations related to EV isolation and characterization methods can lead to misleading or contradictory findings. Here, we examined changes in plasma EVs after mouse SCI at multiple timepoints (1d, 3d, 7d, 14d) using complementary measurement techniques. Plasma EVs isolated by ultracentrifugation (UC) were decreased at 1d post-injury, as shown by nanoparticle tracking analysis (NTA) and paralleled an overall reduction in total plasma extracellular nanoparticles. Western blot (WB) analysis of UC-derived plasma EVs revealed increased expression of the tetraspanin exosome marker, CD81, between 1d and 7d post-injury. To substantiate these findings, we performed interferometric and fluorescence imaging of single, tetraspanin EVs captured directly from plasma with ExoView®. Consistent with WB, we observed significantly increased plasma CD81+ EV count and cargo at 1d post-injury. The majority of these tetraspanin EVs were smaller than 50nm based on interferometry and were insufficiently resolved by flow cytometry-based detection. At the injury site, there was enhanced expression of EV biogenesis proteins that were also detected in EVs directly isolated from spinal cord tissue by WB. Surface expression of tetraspanins CD9 and CD63 increased in multiple cell types at the injury site; however, astrocyte CD81 expression uniquely decreased, as demonstrated by flow cytometry. UC-isolated plasma EV microRNA cargo was also significantly altered at 1d post-injury, with changes similar to that reported in EVs released by astrocytes after inflammatory stimulation. When injected into the lateral ventricle, plasma EVs from SCI mice increased both pro- and anti-inflammatory gene as well as reactive astrocyte gene expression in brain cortex. These studies provide the first detailed characterization of plasma EV dynamics after SCI and suggest that plasma EVs may be involved in posttraumatic brain inflammation.
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影响因子：
16
作者：
Crescitelli R;Lässer C;Szabó TG;Kittel A;Eldh M;Dianzani I;Buzás EI;Lötvall J
通讯作者：
Lötvall J
DOI：
10.1111/jth.12767
发表时间：
2015-02
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Arraud N;Gounou C;Linares R;Brisson AR
通讯作者：
Brisson AR
影响因子：
9.3
作者：
Das M;Mohapatra S;Mohapatra SS
通讯作者：
Mohapatra SS
影响因子：
2.2
作者：
Baek, Rikke;Sondergaard, Evo K. L.;Jorgensen, Malene M.
通讯作者：
Jorgensen, Malene M.
Let-7d、Let-7g 和 Let-7i 的组合可作为血清 microRNA 标准化的稳定参考
影响因子：
3.7
作者：
Chen X;Liang H;Guan D;Wang C;Hu X;Cui L;Chen S;Zhang C;Zhang J;Zen K;Zhang CY
通讯作者：
Zhang CY