Du Jinliang, Jia Rui, Cao Li-Ping, Ding Weidong, Xu Pao, Yin Guojun
Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China.
International Joint Research Laboratory for Fish Immunopharmacology, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China.
Fish Physiol Biochem. 2018 Jun;44(3):747-768. doi: 10.1007/s10695-017-0428-2. Epub 2018 Mar 30.
Fatty liver is an increasingly serious disease of fish in aquaculture. However, the mechanisms responsible for the occurrence of fatty liver remain unclear, and no effective methods for the prevention and treatment of this disease have yet been found. In the present study, we aimed to develop an in vitro model of hepatocyte injury using oleic acid as hepatotoxicant and evaluate the protective effects of Rhizoma Alismatis extract (RAE) in Jian carp using this model. Primary hepatocytes from Jian carp were isolated and purified and cultured in vitro. The result indicated that 0.4 mmol L oleic acid and 48 h could be the optimal conditions to induce hepatocyte injury model in cultured hepatocytes. Hepatocytes were exposed to oleic acid, followed by the addition of RAE at 0, 1, 5, 10, 20, or 50 μg mL. The hepatocytes and supernatant were then analyzed. RAE suppressed oleic acid-induced elevations in aspartate aminotransferase, alanine aminotransferase, triglycerides, total cholesterol, lactate dehydrogenase, alkaline phosphatase, cholinesterase, malondialdehyde, γ-glutamyl transferase, cytochrome P450 1A, cytochrome P450 2E1, liver-type fatty acid binding protein, free fatty acid, fatty acid synthetase, and tumor necrosis factor-α (P < 0.01 or P < 0.05); reduced protein levels of cytochrome P450 1A, nuclear factor (NF)-κB p65, and NF-κB c-Rel; and inhibited cytochrome P4503A, NF-κB c-Rel, nuclear factor erythroid-related factor 2, peroxisome proliferator-activated receptor-α, and cytochrome P4501A mRNA levels. In conclusion, RAE exhibited a protective effect against hepatocyte injury in Jian carp. Further in vivo studies are needed to provide more evidence for the use of RAE as a hepatoprotective agent for the treatment of hepatocyte injury.
脂肪肝是水产养殖中鱼类日益严重的一种疾病。然而，脂肪肝发生的机制仍不清楚，尚未找到预防和治疗该病的有效方法。在本研究中，我们旨在以油酸作为肝毒性剂建立体外肝细胞损伤模型，并使用该模型评估泽泻提取物（RAE）对建鲤的保护作用。分离并纯化建鲤原代肝细胞，然后进行体外培养。结果表明，0.4 mmol/L油酸和48小时可能是诱导培养肝细胞损伤模型的最佳条件。将肝细胞暴露于油酸中，然后分别添加0、1、5、10、20或50 μg/mL的RAE。随后对肝细胞和上清液进行分析。RAE可抑制油酸诱导的天冬氨酸转氨酶、丙氨酸转氨酶、甘油三酯、总胆固醇、乳酸脱氢酶、碱性磷酸酶、胆碱酯酶、丙二醛、γ-谷氨酰转移酶、细胞色素P450 1A、细胞色素P450 2E1、肝型脂肪酸结合蛋白、游离脂肪酸、脂肪酸合成酶和肿瘤坏死因子-α升高（P<0.01或P<0.05）；降低细胞色素P450 1A、核因子（NF）-κB p65和NF-κB c-Rel的蛋白水平；并抑制细胞色素P4503A、NF-κB c-Rel、核因子红系相关因子2、过氧化物酶体增殖物激活受体-α和细胞色素P4501A的mRNA水平。总之，RAE对建鲤肝细胞损伤具有保护作用。需要进一步的体内研究为将RAE用作治疗肝细胞损伤的肝保护剂提供更多证据。
