与使用电子烟相关的内皮细胞衍生的细胞外囊泡会损害脑血管细胞功能。
Endothelial-derived extracellular vesicles associated with electronic cigarette use impair cerebral microvascular cell function.
机构信息
Integrative Vascular Biology Laboratory, Department of Integrative Physiology, University of Colorado Boulder, Boulder, Colorado, United States.
School of Kinesiology, University of Minnesota, Minneapolis, Minnesota, United States.
出版信息
J Appl Physiol (1985). 2023 Aug 1;135(2):271-278. doi: 10.1152/japplphysiol.00243.2023. Epub 2023 Jun 22.
The aim of this study was to determine the effect of circulating endothelial cell-derived microvesicles (EMVs) isolated from e-cigarette users on human cerebral microvascular endothelial cells (hCMECs) nitric oxide (NO) and endothelin (ET)-1 production and tissue-type plasminogen activator (t-PA) release. Circulating EMVs (CD144-PE) were isolated (flow cytometry) from 27 young adults (19-25 yr): 10 nonsmokers (6 M/4 F), 10 e-cigarette users (6 M/4 F), and 7 tobacco cigarette smokers (4 M/3 F). hCMECs were cultured and treated with isolated EMVs for 24 h. EMVs from e-cigarette users and cigarette smokers induced significantly higher expression of p-eNOS (Thr495; 28.4 ± 4.6 vs. 29.1 ± 2.8 vs. 22.9 ± 3.8 AU), Big ET-1 (138.8 ± 19.0 vs. 141.7 ± 19.1 vs. 90.3 ± 18.8 AU) and endothelin converting enzyme (107.6 ± 10.1 and 113.5 ± 11.8 vs. 86.5 ± 13.2 AU), and significantly lower expression of p-eNOS (Ser1177; 7.4 ± 1.7 vs. 6.5 ± 0.5 vs. 9.7 ± 1.6 AU) in hCMECs than EMVs from nonsmokers. NO production was significantly lower and ET-1 production was significantly higher in hCMECs treated with EMVs from e-cigarette (5.7 ± 0.8 µmol/L; 33.1 ± 2.9 pg/mL) and cigarette smokers (6.3 ± 0.7 µmol/L; 32.1 ± 3.9 pg/mL) than EMVs from nonsmokers (7.6 ± 1.2 µmol/L; 27.9 ± 3.1 pg/mL). t-PA release in response to thrombin was significantly lower in hCMECs treated with EMVs from e-cigarette users (from 38.8 ± 6.3 to 37.4 ± 8.3 pg/mL) and cigarette smokers (31.5 ± 5.5 to 34.6 ± 8.4 pg/mL) than EMVs from nonsmokers (38.9 ± 4.3 to 48.4 ± 7.9 pg/mL). There were no significant differences in NO, ET-1, or t-PA protein expression or production in hCMECs treated with EMVs from e-cigarette users and smokers. Circulating EMVs associated with e-cigarette use adversely affects brain microvascular endothelial cells and may contribute to reported cerebrovascular dysfunction with e-cigarette use. In the present study, we determined the effect of circulating endothelial cell-derived microvesicles (EMVs) isolated from e-cigarette users on human cerebral microvascular endothelial cells (hCMECs) nitric oxide (NO) and endothelin (ET)-1 production and tissue-type plasminogen activator (t-PA) release. EMVs from e-cigarette users reduced brain microvascular endothelial cell NO production, enhanced ET-1 production, and impaired endothelial t-PA release. EMVs are a potential mediating factor in the increased risk of stroke associated with e-cigarette use.
本研究旨在确定从电子烟使用者中分离出的循环内皮细胞衍生的微泡（EMVs）对人脑血管内皮细胞（hCMECs）一氧化氮（NO）和内皮素（ET）-1产生和组织型纤溶酶原激活物（t-PA）释放的影响。从 27 名年轻成年人（19-25 岁）中分离出循环 EMVs（CD144-PE）：10 名非吸烟者（6 名男性/4 名女性），10 名电子烟使用者（6 名男性/4 名女性）和 7 名香烟吸烟者（4 名男性/3 名女性）。培养 hCMECs 并用分离的 EMVs 处理 24 小时。电子烟使用者和香烟吸烟者的 EMVs 诱导 p-eNOS（Thr495）表达显著升高（28.4±4.6 与 29.1±2.8 与 22.9±3.8 AU），Big ET-1（138.8±19.0 与 141.7±19.1 与 90.3±18.8 AU）和内皮素转换酶（107.6±10.1 和 113.5±11.8 与 86.5±13.2 AU），而 p-eNOS（Ser1177）表达显著降低（7.4±1.7 与 6.5±0.5 与 9.7±1.6 AU）。与非吸烟者相比，电子烟和香烟吸烟者 EMVs 处理的 hCMECs 中 NO 产生显著降低，ET-1 产生显著升高。与非吸烟者（7.6±1.2 µmol/L；27.9±3.1 pg/mL）相比，电子烟（5.7±0.8 µmol/L；33.1±2.9 pg/mL）和香烟吸烟者（6.3±0.7 µmol/L；32.1±3.9 pg/mL）的 hCMECs 中 t-PA 释放显著降低。与非吸烟者（38.9±4.3 至 48.4±7.9 pg/mL）相比，电子烟使用者（38.8±6.3 至 37.4±8.3 pg/mL）和香烟吸烟者（31.5±5.5 至 34.6±8.4 pg/mL）的 hCMECs 中 t-PA 释放明显降低。与电子烟和香烟使用者的 EMVs 处理的 hCMECs 中，NO、ET-1 或 t-PA 蛋白表达或产生均无显著差异。与电子烟使用相关的循环内皮细胞衍生的微泡（EMVs）对脑微血管内皮细胞产生不利影响，并可能导致与电子烟使用相关的报道的脑血管功能障碍。在本研究中，我们确定了从电子烟使用者中分离出的循环内皮细胞衍生的微泡（EMVs）对人脑血管内皮细胞（hCMECs）一氧化氮（NO）和内皮素（ET）-1产生和组织型纤溶酶原激活物（t-PA）释放的影响。电子烟使用者的 EMVs 降低了脑微血管内皮细胞的 NO 产生，增强了 ET-1 的产生，并损害了内皮细胞的 t-PA 释放。EMVs 是与电子烟使用相关的中风风险增加的潜在介导因素。
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