建鲤急性肝损伤模型的建立及当归提取物的保肝和抗氧化作用研究

摘要

通过四氯化碳(CCl4)诱导建鲤Cyprinus carpio var.Jian肝损伤并建立急性肝损伤模型,以研究当归提取物(Extract from Angelica sinennsis,EAS)对肝脏组织的保护和抗氧化作用.采用CCl4腹腔注射法造模,分别测定不同时间和剂量下建鲤血清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)的活性及丙二醛(MDA)的含量,确定出建模的合适注射浓度和作用时间;用添加0.5％和1.0％(质量分数,下同)的EAS饲料,预防性喂养建鲤60 d后,通过测定鲤血清和肝脏组织中相关生化指标来观察EAS的保肝和抗氧化作用.结果表明:用体积分数为15.00％的CCl4-橄榄油一次性腹腔注射(剂量为0.1 mL/10 g体质量)建鲤72 h后,能诱导鲤血清中GPT、GOT和LDH活性显著性升高(P＜0.05),MDA大量生成;投喂含0.5％和1.0％的EAS饲料均能显著抑制CCl4致建鲤血清中GOT、GPT和LDH活性的升高(P＜0.05),提高血清中总蛋白(TP)、白蛋白(Alb)含量及肝组织总抗氧化能力(T-AOC),抑制肝组织匀浆中超氧化物歧化酶(SOD)水平下降;同时,0.5％的EAS还能显著恢复血清中SOD水平和肝组织中谷胱甘肽(GSH)水平,抑制MDA生成.本研究结果表明,EAS对CCl4诱导的建鲤肝组织损伤具有保护作用,该作用与EAS的清除自由基和抗氧化能力有关.%The present study aimed to develop a vivo model of acute hepatotoxicity using carbon tetrachloride ( CCl4) as hepatotoxicant and to evaluate the antioxidant effects of an extract from Angelica sinennsis ( EAS ) against CCl4 induced hepatotoxicity in Jian common carp ( Cyprinus carpio var. Jian). The Jian common carp model of a-cute hepatic injury was induced by intraperitoneal injection of CCl4. To define the most optimal CCl4 action concen-tration and time, the serum glutamate oxalate transaminase( GOT) ,glutamate pyruvate transaminase ( GPT) ,lactate dehydrogenase(LDH) and malondialdehyde ( MDA) were detected at different time and doses. The Jian common carp was fed the diet containing EAS at a dose of 0. 5 and 1 g/100 g for 60 days prior to CCl4 intoxication, and the changes in hepatic function markers in serum and hepatic tissue were analyzed to evaluate the antioxidant effects of EAS on the hepatopancrease. Results showed that the serum level of MDA and GOT,GPT,and LDH activities were found to be significantly elevated in the group injected with 15. 00% (in volume) of CC14 in olive oil at a volume of 0. 1 mL/10 g of body weight for 72 h. In vivo, administration of EAS at the doses of 0. 5 and 1 g/100 g in the diet for 6 0 days led to significant reduce in the activities of GPT, GOT and LDH and increase in the reduced levels of total protein (TP) and albumin ( Alb) in the serum. Meanwhile, the reduced levels of superoxide dismutase (SOD) ,glutathione ( GSH) and total antioxidant capacity (T-AOC) were markedly increased and the MDA forma-tion was significantly inhibited in the hepatopancrease. Fish received 0. 5 g/100 g of EAS in the diets showed sig-nificant restoration of SOD activities in blood serum and glutathione( GSH) in liver homogenate, and reduction of MDA. The findings proved the hepatoprotective action of EAS, which is likely related to antioxidative activity, and support the use of EAS as a hepatoprotective and antioxidant agent in fish.