邹翠云, 陈小晶, 邬颖欣, 黄锦雄, 谭小红, 胡心月, 甘松永, 吴锦辉. 小柴胡汤对D-GalN/LPS诱导的虎龙杂交石斑鱼肝细胞损伤的保护作用[J]. 南方水产科学, 2022, 18(5): 66-73. DOI: 10.12131/20210362
引用本文: 邹翠云, 陈小晶, 邬颖欣, 黄锦雄, 谭小红, 胡心月, 甘松永, 吴锦辉. 小柴胡汤对D-GalN/LPS诱导的虎龙杂交石斑鱼肝细胞损伤的保护作用[J]. 南方水产科学, 2022, 18(5): 66-73. DOI: 10.12131/20210362
ZOU Cuiyun, CHEN Xiaojing, WU Yingxin, HUANG Jinxiong, TAN Xiaohong, HU Xinyue, GAN Songyong, WU Jinhui. Protective effect of Xiaochaihu Decoction on D-GalN/ LPS-induced hepatocyte injury in hybrid grouper (Epinephelus lanceolatusy♂× E. fuscoguttatus♀)[J]. South China Fisheries Science, 2022, 18(5): 66-73. DOI: 10.12131/20210362
Citation: ZOU Cuiyun, CHEN Xiaojing, WU Yingxin, HUANG Jinxiong, TAN Xiaohong, HU Xinyue, GAN Songyong, WU Jinhui. Protective effect of Xiaochaihu Decoction on D-GalN/ LPS-induced hepatocyte injury in hybrid grouper (Epinephelus lanceolatusy♂× E. fuscoguttatus♀)[J]. South China Fisheries Science, 2022, 18(5): 66-73. DOI: 10.12131/20210362

小柴胡汤对D-GalN/LPS诱导的虎龙杂交石斑鱼肝细胞损伤的保护作用

Protective effect of Xiaochaihu Decoction on D-GalN/ LPS-induced hepatocyte injury in hybrid grouper (Epinephelus lanceolatusy♂× E. fuscoguttatus♀)

  • 摘要: 为探讨小柴胡汤对虎龙杂交石斑鱼 (Epinephelus lanceolatus♂×E. fuscoguttatus♀) 化学性肝细胞损伤的免疫功能影响,用D-氨基半乳糖 (D-GalN, 20 mmol·L–1) 和脂多糖 (LPS, 1 μg·mL–1) 作用肝细胞24 h构建虎龙杂交石斑鱼肝细胞损伤模型,用不同质量浓度的小柴胡汤 (100、200和400 μg·mL–1) 对肝细胞进行前处理 D-氨基半乳糖联合脂多糖 (D-GalN/LPS) 损伤前加药,通过CCK-8法检测肝细胞活力,HE染色观察肝细胞形态,收集细胞上清液检测炎症因子INF-γ、COX-2和PGE2的含量,以及细胞沉淀中凋亡和免疫相关基因的表达。结果表明,小柴胡汤可以提高肝细胞的成活率,不同程度地抑制D-GalN/LPS损伤引起的肝细胞培养上清液中INF-γ活性以及COX-2和PGE2含量的升高,降低细胞内乳酸脱氢酶 (LDH)、谷草转氨酶 (AST) 和谷丙转氨酶 (ALT) 活性以及细胞凋亡率,对肝细胞结构具有明显的保护作用;同时显著抑制凋亡相关基因caspase-3、caspase-9和P53的上调,促进免疫相关基因TLR3的上调。数据统计分析显示,添加200 μg·mL–1小柴胡汤组的抑制效果最明显。综合以上结果,小柴胡汤对虎龙杂交石斑鱼化学性肝损伤有一定的保护作用,可预防肝脏疾病的发生和发展,为鱼类保肝药物研究提供了新的见解。

     

    Abstract: The study aims to evaluate the protective effect of Xiaochaihu Decoction (XCHD) on chemical hepatocyte injury. We treated the hepatocytes with D-GalN (20 mmol·L–1) and LPS (1 μg·mL–1) for 24 h to build a hybrid grouper (Epinephelus lanceolatus♂×E. fuscoguttatus♀) hepatocyte injury model, then pretreated the liver cells with different concentrations of XCHD (100, 200 and 400 μg·mL–1) (D-GalN/LPS damage), and measured the hepatocyte viability by the CCK-8 assay. Hepatocyte morphology was visualized by HE staining, and cell supernatants were collected to detect the content of the inflammatory factors INF-, COX-2, and PGE2, as well as the expression of apoptotic and immune-related genes in the cell cryoprecipitate. The results show that XCHD can improve the survival rate of hepatocytes, inhibit INF-γ activity and coX-2 and PGE2 contents in the supernatant of hepatocytes induced by D-GalN/LPS injury to a different extent, reduce LDH, AST and ALT contents and apoptosis rate of hepatocytes. Besides, it had a significant protective effect on the structure of hepatocytes, significantly inhibiting the up-regulation of apoptosis-related genes caspase-3, caspase-9 and P53, and enhancing the up-regulation of immune-related genes TLR3. According to the data from the statistical analysis, the inhibition effect was most obvious in the 200 μg·mL–1 XCHD group. In conclusion, XCHD has a protective effect on the chemical hepatocyte injury of Epinephelus lanceolatus♂×E. fuscoguttatus♀, and can prevent the occurrence and development of liver disease, which provides a new insight for the study of liver protection drugs in fish.

     

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