Increased liver-specific proteins in circulating extracellular vesicles as potential biomarkers for drug- and alcohol-induced liver injury.
Increased liver-specific proteins in circulating extracellular vesicles as potential biomarkers for drug- and alcohol-induced liver injury.
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DOI：
10.1371/journal.pone.0172463
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Baek MC
中科院分区：
文献类型：
--
作者：
Cho YE;Im EJ;Moon PG;Mezey E;Song BJ;Baek MC
药物和酒精引起的肝损伤是肝衰竭和肝移植的主要原因。新出现的证据表明，细胞外囊泡（EV）是生物标志物的来源，因为它们含有反映EV蛋白的身份和组织特异性起源的独特蛋白质。这项研究旨在确定潜在的肝毒性药物，如对乙酰氨基酚（APAP）和酗酒，是否可以增加循环EV的数量，并评估肝脏特异性EV蛋白作为肝损伤的潜在生物标志物。从APAP暴露、乙醇喂养的小鼠或酒精性肝炎患者与正常对照组的血浆中分离的循环EV，通过蛋白质组学和生化方法进行表征。通过免疫印迹和ELISA分析肝特异性EV蛋白。APAP处理小鼠的循环EV中总蛋白和肝脏特异性蛋白的量以剂量和时间依赖性方式显著增加。来自APAP暴露小鼠的EV的蛋白质组学分析显示，与对照组相比，肝脏特异性和/或肝毒性蛋白的量增加。此外，当用N-乙酰半胱氨酸或谷胱甘肽处理小鼠时，APAP暴露后EV中增加的蛋白质量恢复到基础水平。在暴露于肝毒性剂量的硫代乙酰胺或d-半乳糖胺的小鼠中也观察到循环EV中的量和肝脏特异性蛋白质增加的类似结果，但非肝毒性青霉素或肌毒性布比卡因未观察到。此外，与对照组相比，酗酒暴露显著升高了患有酒精性肝炎的小鼠和酗酒者的循环EV中的肝脏特异性蛋白质。这些结果表明，药物和酒精介导的肝损伤中的循环EV含有肝脏特异性蛋白，可以作为肝毒性的特异性生物标志物。
Drug- and alcohol-induced liver injury are a leading cause of liver failure and transplantation. Emerging evidence suggests that extracellular vesicles (EVs) are a source of biomarkers because they contain unique proteins reflecting the identity and tissue-specific origin of the EV proteins. This study aimed to determine whether potentially hepatotoxic agents, such as acetaminophen (APAP) and binge alcohol, can increase the amounts of circulating EVs and evaluate liver-specific EV proteins as potential biomarkers for liver injury. The circulating EVs, isolated from plasma of APAP-exposed, ethanol-fed mice, or alcoholic hepatitis patients versus normal control counterparts, were characterized by proteomics and biochemical methods. Liver specific EV proteins were analyzed by immunoblots and ELISA. The amounts of total and liver-specific proteins in circulating EVs from APAP-treated mice significantly increased in a dose- and time-dependent manner. Proteomic analysis of EVs from APAP-exposed mice revealed that the amounts of liver-specific and/or hepatotoxic proteins were increased compared to those of controls. Additionally, the increased protein amounts in EVs following APAP exposure returned to basal levels when mice were treated with N-acetylcysteine or glutathione. Similar results of increased amounts and liver-specific proteins in circulating EVs were also observed in mice exposed to hepatotoxic doses of thioacetamide or d-galactosamine but not by non-hepatotoxic penicillin or myotoxic bupivacaine. Additionally, binge ethanol exposure significantly elevated liver-specific proteins in circulating EVs from mice and alcoholics with alcoholic hepatitis, compared to control counterparts. These results indicate that circulating EVs in drug- and alcohol-mediated hepatic injury contain liver-specific proteins that could serve as specific biomarkers for hepatotoxicity.