论文部分内容阅读
以肝(细胞)损伤为主要特征的鱼类肝胆综合症是水产养殖中日趋严重的病害之一,目前还没有有效的防治措施。本研究拟以叔丁基氢过氧化物(t-BHP)构建建鲤(Cyprinus carpio var.jian)原代肝细胞损伤模型,并利用该模型评价甘草(Glycyrrhiza glabra)提取物对t-BHP诱导的鱼类急性肝细胞损伤的保护作用。1mmol/L的t-BHP与原代肝细胞共培养2h能显著提高肝细胞培养上清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)和丙二醛(MDA)水平,显著降低谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)的含量以及肝细胞增殖活性。在t-BHP诱导肝细胞损伤前(前处理)、损伤后(后处理)、损伤前和损伤后(前后处理)将不同浓度(0.1、0.2和0.4mg/mL)的甘草提取物加入肝细胞培养液中,与肝细胞共培养2h,结果显示,前后处理时,不同浓度(0.1、0.2和0.4mg/mL)的甘草提取物均能显著抑制t-BHP诱导的GOT、GPT、LDH和MDA水平的升高,恢复GSH-Px和SOD水平;前处理时,高浓度(0.4mg/mL)的甘草提取物对抑制GOT、GPT、LDH和MDA水平的升高,恢复GSH-Px水平有显著效果;后处理时,只有高浓度(0.4mg/mL)的甘草提取物能有效提高GSH-Px活性;中浓度和高浓度(0.2和0.4mg/mL)的甘草提取物在前处理、后处理及前后处理时均能显著提高肝细胞的增殖活力。研究的结果表明,中药与损伤剂的给予顺序影响着甘草提取物对肝细胞的保护作用,前后处理时甘草提取物对损伤肝细胞的保护效果明显优于前处理和后处理。研究证实了甘草提取物对t-BHP诱导的鱼类肝细胞损伤具有保护作用,对应用甘草提取物作为鱼类肝胆综合症的防治药物还需要进一步的在体研究。
Fish hepatobiliary syndrome characterized by hepatic (cellular) injury is one of the increasingly serious diseases in aquaculture. There is currently no effective prevention and treatment measures. In this study, the primary hepatocyte injury model of Cyprinus carpio var. Jian was constructed by tert-butyl hydroperoxide (t-BHP). The model was used to evaluate the effects of Glycyrrhiza glabra extract on t-BHP- Acute hepatotoxicity. Co-culture of 1 mmol / L t-BHP with primary hepatocytes for 2 h significantly increased the levels of GPT, GOT, LDH and MDA in the supernatant of hepatocytes, Levels, significantly reduce the content of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and hepatocyte proliferation activity. Different concentrations of glycyrrhiza extracts (0.1, 0.2 and 0.4 mg / mL) were added to hepatocytes before t-BHP-induced hepatocellular injury (pretreatment), post injury (posttreatment), pre-injury and post injury The results showed that glycyrrhiza extracts with different concentrations (0.1, 0.2 and 0.4 mg / mL) could significantly inhibit t-BHP-induced GOT, GPT, LDH and MDA The levels of GSH-Px and SOD were restored. The levels of GOT, GPT, LDH and MDA were increased and the level of GSH-Px was restored after pretreatment at high concentration (0.4mg / mL) The results showed that only the high concentration (0.4mg / mL) of licorice extract could effectively increase the activity of GSH-Px in the post-treatment. The medium and high concentration (0.2 and 0.4mg / mL) And before and after treatment can significantly improve the proliferation of liver cells. The results showed that the order of administration of Chinese medicine and damaging agent affected the protective effect of licorice extract on hepatocytes. The protective effect of licorice extract on injured hepatocytes was significantly better than that of pre-treatment and post-treatment. The study confirmed that licorice extract has protective effect on t-BHP-induced fish liver cell injury, and further studies on the use of licorice extract as a preventive and therapeutic drug against fish hepatobiliary syndrome are needed.