Characterization of heat shock protein 27 in extracellular vesicles: a potential anti-inflammatory therapy
Characterization of heat shock protein 27 in extracellular vesicles: a potential anti-inflammatory therapy
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DOI：
10.1096/fj.201800987r
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发表时间：
2019-02-01
期刊：
影响因子：
4.8
通讯作者：
O'Brien, Edward R.
中科院分区：
文献类型：
--
作者：
Shi, Chunhua;Ulke-Lemee, Annegret;O'Brien, Edward R.
此前，我们报道过，血清热休克蛋白 27 (HSP27) 水平升高可预测心脏病发作、中风或心血管疾病死亡的风险较低。此外，增强HSP27（或鼠类直系同源物HSP25）可减弱实验性动脉粥样硬化形成、减少炎症并降低胆固醇水平。最近，我们注意到 HSP27 通过 TLR-4 激活 NF-B，从而减轻斑块炎症；然而，细胞外 HSP27 介导的精确抗动脉粥样硬化机制尚不完全清楚。我们本研究的目的是研究细胞外囊泡 (EV) 中 HSP27 的存在以及 HSP27 是否对靶细胞产生动脉粥样硬化保护作用。在这里，我们使用透射电子显微镜 (TEM) 和免疫金标记、蛋白质印迹、ELISA 和荧光激活细胞分选提供 HSP27 定位于 THP-1 细胞衍生的 EV 的证据。 TEM 成像表明 HSP27 存在于外泌体膜上。对大囊泡（包括从人血浆中分离出的微粒和外泌体）进行多反应器监测-质谱分析，也使用独特的特征肽 R.LFDQAFGLPR.L 检测到 HSP27。使用 THP-1 和人胚胎肾细胞的研究表明，载有 HSP27 的外泌体显着刺激 NF-B 激活 (P < 0.001) 和 IL-10 释放 (P < 0.0001)，表明 HSP27 可能是具有有益抗炎作用的重要外泌体货物。Shi, C.、Ulke-Lemee, A.、Deng, J.、Batulan, Z.、O'Brien, E. R. 细胞外囊泡中热休克蛋白 27 的表征：一种潜在的抗炎疗法。
Previously, we reported that elevated serum levels of heat shock protein 27 (HSP27) are predictive of a lower risk of having a heart attack, stroke, or death from cardiovascular disease. Moreover, augmenting HSP27 (or the murine ortholog, HSP25) attenuated experimental atherogenesis, reduced inflammation, and lowered cholesterol levels. Recently, we noted that HSP27 activates NF-B via TLR-4, resulting in attenuation of plaque inflammation; however, the precise anti-atherosclerosis mechanisms mediated by extracellular HSP27 are incompletely understood. Our purpose in this study was to investigate the existence of HSP27 in extracellular vesicles (EVs) and whether HSP27 elicited atheroprotective effects on target cells. Here, we provide evidence that HSP27 localizes to EVs derived from THP-1 cells using transmission electron microscopy (TEM) and immunogold labeling, Western blotting, ELISA, and fluorescence-activated cell sorting. TEM imaging indicated that HSP27 is found at the exosomal membrane. Multiple reactor monitor-mass spectrometric analysis of large vesicles, which included microparticles and exosomes, isolated from human plasma, also led to detection of HSP27 using the unique signature peptide, R.LFDQAFGLPR.L. Studies using THP-1 and human embryonic kidney cells show that HSP27-laden exosomes significantly stimulated NF-B activation (P < 0.001) and release of IL-10 (P < 0.0001), suggesting that HSP27 may be important exosomal cargo with beneficial anti-inflammatory effects.Shi, C., Ulke-Lemee, A., Deng, J., Batulan, Z., O'Brien, E. R. Characterization of heat shock protein 27 in extracellular vesicles: a potential anti-inflammatory therapy.