The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Hum Exp Toxicol. 2010 Apr;29(4):303-9. doi: 10.1177/0960327110361757. Epub 2010 Feb 9.
Clivorine is an otonecine-type hepatotoxic pyrrolizidine alkaloid (HPAs), to which humans are exposed when consuming herbs containing such components. In the present study, we investigated clivorine-induced oxidative stress injury on primary cultured rat hepatocytes. Rat hepatocytes were treated with various concentrations of clivorine (1-100 microM) for 48 hours, and then cell viability was detected by 3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay, while lipid peroxidation (LPO) level, glutathione peroxidase (GPx), glutathione-S-transferase (GST), glutathione reductase (GR), catalase (CAT) and superoxide dismutase (SOD) activities were determined to evaluate the oxidative injury. The results of MTT assay showed that clivorine decreased cell viability in a concentration-dependent manner. Clivorine also increased LPO amounts in rat hepatocytes at the concentrations of 50 microM and 100 microM. Further results showed that clivorine decreased GPx, GST and GR activities, which are all reduced glutathione (GSH)-related antioxidant enzymes. CAT and SOD are both important antioxidant enzymes, and the results showed that clivorine increased CAT activity at the low concentration of 5 muM and decreased cellular SOD activity at all concentrations. Taken together, our results demonstrated that clivorine induced toxicity on primary cultured rat hepatocytes by causing the damage on cellular redox balance.
克利弗林是一种奥替诺型肝毒性吡咯里西啶生物碱（HPAs），当人类食用含有此类成分的草药时，就会接触到克利弗林。在本研究中，我们研究了克利弗林对原代培养大鼠肝细胞的氧化应激损伤作用。用不同浓度的克利弗林（1-100 μM）处理大鼠肝细胞 48 小时，然后通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测法检测细胞活力，同时测定脂质过氧化（LPO）水平、谷胱甘肽过氧化物酶（GPx）、谷胱甘肽-S-转移酶（GST）、谷胱甘肽还原酶（GR）、过氧化氢酶（CAT）和超氧化物歧化酶（SOD）活性，以评估氧化损伤。MTT 检测结果表明，克利弗林以浓度依赖的方式降低细胞活力。克利弗林还增加了大鼠肝细胞中 50 μM 和 100 μM 浓度的 LPO 含量。进一步的结果表明，克利弗林降低了 GPx、GST 和 GR 活性，这三种酶都是谷胱甘肽（GSH）相关的抗氧化酶。CAT 和 SOD 都是重要的抗氧化酶，结果表明，克利弗林在 5 μM 的低浓度下增加了 CAT 活性，而在所有浓度下都降低了细胞 SOD 活性。总之，我们的结果表明，克利弗林通过破坏细胞氧化还原平衡对原代培养大鼠肝细胞造成毒性。
