姜黄素预处理外泌体减轻高同型半胱氨酸血症中的内皮细胞功能障碍。
Curcumin-primed exosomes mitigate endothelial cell dysfunction during hyperhomocysteinemia.
机构信息
Department of Physiology and Biophysics, School of Medicine, University of Louisville, Louisville, KY 40202, USA.
Department of Physiology and Biophysics, School of Medicine, University of Louisville, Louisville, KY 40202, USA.
出版信息
Life Sci. 2014 Jun 27;107(1-2):1-7. doi: 10.1016/j.lfs.2014.04.018. Epub 2014 Apr 26.
AIM
Exosomes, the nano-units (<200 nm), released from diverse cell types in the extracellular body fluid, possess non-immunogenic property and ability to cross the blood-brain barrier (BBB). Since exosomes carry biological information from their cells of origin, we hypothesize that priming cells with potential therapeutic agents release improved cellular contents through exosomes. Curcumin possesses anti-oxidative and anti-inflammatory properties and provides a promising treatment for cerebral diseases and therefore, the aim of the study is to establish that mouse brain endothelial cells (MBECs) when primed with curcumin (7.5 μM), release an alleviated exosome population that can help recover the endothelial cell (EC) layer permeability.
MAIN METHODS
Homocysteine is a well-known causative factor of BBB disruption; therefore, homocysteine-treated ECs were used as a model of BBB disruption and curcumin-primed exosomes were utilized to check their potential for mitigating EC disruption. MBECs were treated with curcumin and exosomes were isolated by using ultracentrifugation and immunoprecipitation. Expression levels of junction proteins were detected by Western blot and immunocytochemistry assays. Endothelial cell permeability was analyzed with Fluorescein isothiocyanate-Bovine serum albumin (FITC-BSA) leakage assay using transwell permeable supports.
KEY FINDINGS
Exosomes derived from curcumin-treated (primed) cells (CUR-EXO) alleviated oxidative stress, tight junctions (ZO-1, claudin-5, occludin), adherent junction (VE-cadherin) proteins and EC layer permeability induced during EC damage due to high homocysteine levels (hyperhomocysteinemia).
SIGNIFICANCE
In conclusion, the study potentiates the use of CUR-EXO for cerebral diseases where drug delivery is still a challenge. The results also pave the way to novel translational therapies for cerebral diseases by maintaining and establishing therapeutic conservatories via primed exosomes.
目的
外泌体是细胞在细胞外体液中释放的纳米级（<200nm）小单位，具有非免疫原性和穿越血脑屏障（BBB）的能力。由于外泌体携带了其起源细胞的生物信息，我们假设用潜在治疗药物预处理细胞可以通过外泌体释放改善的细胞内容物。姜黄素具有抗氧化和抗炎特性，为脑部疾病提供了一种有前途的治疗方法，因此，本研究的目的是确定用姜黄素（7.5μM）预处理的小鼠脑内皮细胞（MBECs）释放的减轻的外泌体群体可以帮助恢复内皮细胞（EC）层通透性。
主要方法
同型半胱氨酸是 BBB 破坏的已知原因；因此，用同型半胱氨酸处理的 EC 用作 BBB 破坏的模型，并利用姜黄素预处理的外泌体来检查其减轻 EC 破坏的潜力。用姜黄素处理 MBECs，并通过超速离心和免疫沉淀分离外泌体。通过 Western blot 和免疫细胞化学检测连接蛋白的表达水平。使用 Transwell 可渗透支持物，通过荧光素异硫氰酸酯-BSA（FITC-BSA）渗漏测定法分析内皮细胞通透性。
主要发现
来自姜黄素处理（预处理）细胞的外泌体（CUR-EXO）减轻了由于高同型半胱氨酸水平（高同型半胱氨酸血症）引起的氧化应激、紧密连接（ZO-1、claudin-5、occludin）、粘着连接（VE-钙粘蛋白）蛋白和 EC 层通透性受损。
意义
总之，该研究促进了 CUR-EXO 在药物输送仍然是一个挑战的脑部疾病中的应用。研究结果还为通过维持和建立经过预处理的外泌体治疗性保护区，为脑部疾病开辟了新的转化治疗途径。
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