Hubei Key Laboratory of Food Nutrition and Safety, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Henan Provincial Center for Disease Control and Prevention, Zhengzhou, Henan, China.
J Nutr Biochem. 2023 Jun;116:109332. doi: 10.1016/j.jnutbio.2023.109332. Epub 2023 Mar 24.
Alcoholic liver disease (ALD), a spectrum of liver abnormalities induced by chronic alcohol abuse, continues to be the major cause of life-threatening liver disease in developed countries. Autophagy and exosomes were individually confirmed to be involved in the pathogenesis of ALD. Here, we sought to identify the role of autophagy and exosomes in the liver protective effects of quercetin. We observed decreased hepatic LC3II/LC3I and increased p62 level in ethanol-fed mice, and these changes were alleviated by quercetin. Meanwhile, nanoparticle tracking analysis (NTA) showed elevated serum exosomes numbers in ethanol-fed mice, which was combated by quercetin. Ethanol induced elevated LDH, ALT, and AST in HepG2 supernatant, which was alleviated by cytochalasin D (exosomes uptake inhibitor). Moreover, quercetin reduced ethanol-induced LDH and ALT elevation in vitro, and the effects of quercetin were reversed by Rab27a overexpression (induce exosomes release) or wortmannin treatment (autophagy inhibitor). Transcriptomic analysis supported that quercetin reversed the change of lysosome related genes disturbed by ethanol. Meanwhile, western blot analysis exhibited decreased hepatic expression of LAMP2 and ATPA6V1B2, and active Cathepsin B/Cathepsin B by quercetin treatment, indicating quercetin alleviated lysosome dysfunction in ethanol-fed mice. Baf A treatment or transfection of siTFEB offset quercetin's effects in ethanol-induced LDH and ALT elevation, exosomes release, and autophagy inhibition (LC3II/I and p62 accumulation). Taken together, quercetin coordinately activates autophagy and combats exosomes release by restoring lysosome function, and further mitigates ethanol-induced liver damage.
酒精性肝病（ALD）是一种由慢性酒精滥用引起的肝脏异常谱，在发达国家仍然是威胁生命的肝脏疾病的主要原因。自噬和外泌体分别被证实参与了 ALD 的发病机制。在这里，我们试图确定自噬和外泌体在槲皮素肝脏保护作用中的作用。我们观察到乙醇喂养的小鼠肝组织中 LC3II/LC3I 减少，p62 水平增加，而槲皮素可以减轻这些变化。同时，纳米颗粒跟踪分析（NTA）显示乙醇喂养的小鼠血清外泌体数量增加，而槲皮素可以对抗这种增加。乙醇诱导 HepG2 上清液中 LDH、ALT 和 AST 升高，细胞松弛素 D（外泌体摄取抑制剂）可以减轻这种升高。此外，槲皮素在体外减轻了乙醇诱导的 LDH 和 ALT 升高，而 Rab27a 过表达（诱导外泌体释放）或wortmannin 处理（自噬抑制剂）逆转了槲皮素的作用。转录组分析支持槲皮素逆转了乙醇干扰的溶酶体相关基因的变化。同时，Western blot 分析显示，槲皮素处理后肝组织 LAMP2 和 ATPA6V1B2 以及活性 Cathepsin B/Cathepsin B 的表达减少，表明槲皮素减轻了乙醇喂养小鼠的溶酶体功能障碍。Baf A 处理或 siTFEB 转染抵消了槲皮素在乙醇诱导的 LDH 和 ALT 升高、外泌体释放和自噬抑制（LC3II/I 和 p62 积累）中的作用。总之，槲皮素通过恢复溶酶体功能协调激活自噬并对抗外泌体释放，从而进一步减轻乙醇引起的肝损伤。
