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甘草提取物对叔丁基氢过氧化物(t-BHP)诱导的建鲤原代培养肝细胞损伤的保护作用
曹丽萍1,贾睿2,杜金梁2,丁炜东2,殷国俊2
1.
2. 中国水产科学研究院淡水渔业研究中心
Protective Effect of Glycyrrhiza glabra Extract against tert-butyl hydroperoxide(t-BHP)-induced Hepatotoxicity in Primary Cultured Hepatocytes of Jian Carp(Cyprinus carpio var. jian)
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摘要 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。1 mmol/L的 t-BHP与原代肝细胞共培养2 h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0.4 mg/mL)的甘草提取物加入肝细胞培养液中,与肝细胞共培养2 h,结果显示,前后处理时,不同浓度(0.1、0.2和0.4 mg/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(0.4 mg/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(0.4 mg/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。
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曹丽萍
贾睿
杜金梁
丁炜东
殷国俊
关键词 建鲤甘草提取物原代培养模型抗氧化作用    
Abstract:Fish "liver and gall syndrome", characterized by liver (hepatocyte) injury, has become more and more serious in China aquaculture, however no effective methods have been found for the prevention and treatment of this disease. The present study aimed to develop an in vitro model of hepatotoxicity using tert-butyl hydroperoxide (t-BHP) as hepatotoxicant and evaluate the protective effects of Glycyrrhiza glabra extract against t-BHP induced hepatotoxicity in Jian carp (Cyprinus carpio var. jian). Exposure of the hepatocytes to 1 mmol/L t-BHP for 2 h significantly elevated the levels of glutamate oxalate transaminase (GOT), glutamate pyruvate transaminase (GPT), lactate dehydrogenase (LDH) and malondialdehyde (MDA), and reduced the cell viability and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Three concentrations of G. glabra extract (0.1, 0.2 and 0.4 mg/mL) were added to the primary hepatocytes before (pre-treatment), after (post-treatment) and both before and after (pre- and post-treatment) the exposure of the hepatocytes to t-BHP. Results showed that pre- and post-treatment of the hepatocytes with G. glabra extract at 0.1, 0.2 and 0.4 mg/mL suppressed the elevations of LDH, GOT, GPT and MDA, and reversed the reduced activities of GSH-Px and SOD. Pre-treatment of the hepatocytes with 0.4 mg/mL G. glabra extract reduced the increased levels of GPT, GOT, LDH and MDA, and increased the reduced GSH-Px activity. Post-treatment didn't show significant effects expect for the enhanced GSH-Px activity in the cells post-treated with 0.4 mg/mL G. glabra extract. The significantly increased cell viabilities were observed when the cells were pre-treated, post-treated and pre- and post-treated with 0.2 mg/mL and 0.4 mg/mL G. glabra extract. The results also showed that the timing of G. glabra extract treatment substantially influenced the protective efficacy of the extract, the pre- and post-treatment of the hepatocytes with G. glabra extract was better than that of the other two treatment regimes. It can be concluded that G. glabra extract exhibited protective effect against t-BHP-induced hepatotoxicity in fish. Further in vivo studies are needed to provide more evidence for using G. glabra as a hepatoprotective agent for the prevention and treatment of fish "liver and gall syndrome".
Key wordsCyprinus carpio var. jian    Glycyrrhiza glabra extract    Primary culture    Model    Anti-oxidative activity
收稿日期: 2012-02-10     
通讯作者: 殷国俊   
引用本文:   
曹丽萍1,贾睿2,杜金梁2,丁炜东2,殷国俊2. 甘草提取物对叔丁基氢过氧化物(t-BHP)诱导的建鲤原代培养肝细胞损伤的保护作用[J]. , 2012, 20(10): 1192-1200.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I10/1192
 
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