Institute for Liver Diseases of Anhui Medical University, School of Pharmacy, Anhui Medical University, Hefei 230032, China; Key Laboratory of anti-inflammatory and Immune Medicine, Ministry of Education, Hefei 230032, China.
Institute for Liver Diseases of Anhui Medical University, School of Pharmacy, Anhui Medical University, Hefei 230032, China; Key Laboratory of anti-inflammatory and Immune Medicine, Ministry of Education, Hefei 230032, China.
Eur J Pharmacol. 2019 May 5;850:15-22. doi: 10.1016/j.ejphar.2019.02.005. Epub 2019 Feb 10.
Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease which affects millions of people worldwide. Acetaminophen (APAP) overdose is the leading cause of acute liver failure. In this study, APAP (50, 100, 200 mg/kg) were employed on mice fed with a high-fat diet, and APAP (2, 4, 8 mM) were cultured with L02 cells in the presence of alcohol and oleic acid. APAP treatment significantly aggravated hepatic lipid accumulation, increased the serum levels of triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and increased hepatic lipid accumulation in H&E and Oil red O staining results. Transmission electron microscopy (TEM) found fewer number of autophagosomes in APAP (100 mg/kg) treated group. Immunohistochemistry analysis showed the intensity of hepatic mTOR was increased and AMPK was decreased in 200 mg/kg APAP treated group. Western blot analysis showed APAP treatment decreased the levels of LC3-Ⅱ, Beclin1 and AMPK, while increased the levels of mTOR and SREBP-1c, respectively. In vitro study showed APAP treatment obviously increased TG activities in cell supernatant, and Oil red O staining had the same results. Western blot analysis demonstrated APAP treatment decreased the levels of LC3-Ⅱ, Beclin1 and AMPK, increased the levels of mTOR and SREBP-1c, but rapamycin treatment significantly reversed these effects of APAP. In conclusion, therapeutic dosages of APAP aggravates fat accumulation in NAFLD, the potential mechanism might be involved in inhibiting autophagy associated with the AMPK/mTOR pathway, and patients with NAFLD should use a lower dose of APAP.
非酒精性脂肪性肝病（NAFLD）是世界范围内最常见的慢性肝病，影响着数百万人。对乙酰氨基酚（APAP）过量是急性肝衰竭的主要原因。在这项研究中，APAP（50、100、200mg/kg）被用于喂养高脂肪饮食的小鼠，APAP（2、4、8mM）在酒精和油酸存在的情况下与 L02 细胞共培养。APAP 治疗显著加重肝脂质堆积，增加血清甘油三酯（TG）、丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平，并增加 H&E 和油红 O 染色结果中的肝脂质堆积。透射电子显微镜（TEM）发现 APAP（100mg/kg）治疗组中的自噬体数量减少。免疫组化分析显示，200mg/kg APAP 治疗组肝 mTOR 强度增加，AMPK 减少。Western blot 分析显示，APAP 处理降低了 LC3-Ⅱ、Beclin1 和 AMPK 的水平，而分别增加了 mTOR 和 SREBP-1c 的水平。体外研究表明，APAP 处理明显增加细胞上清液中的 TG 活性，油红 O 染色也有相同的结果。Western blot 分析表明，APAP 处理降低了 LC3-Ⅱ、Beclin1 和 AMPK 的水平，增加了 mTOR 和 SREBP-1c 的水平，但雷帕霉素处理显著逆转了 APAP 的这些作用。总之，APAP 的治疗剂量会加重 NAFLD 中的脂肪堆积，其潜在机制可能涉及抑制与 AMPK/mTOR 通路相关的自噬，NAFLD 患者应使用较低剂量的 APAP。
