Injac Rade, Perse Martina, Obermajer Natasa, Djordjevic-Milic Vukosava, Prijatelj Matevz, Djordjevic Aleksandar, Cerar Anton, Strukelj Borut
Faculty of Pharmacy, The Chair of Pharmaceutical Biology, University of Ljubljana, Askerceva 7, 1000 Ljubljana, Slovenia.
Biomaterials. 2008 Aug-Sep;29(24-25):3451-60. doi: 10.1016/j.biomaterials.2008.04.048. Epub 2008 May 27.
The aim of this study was to investigate the potential protective role of fullerenol C60(OH)24 on doxorubicin-induced liver toxicity using in vivo (female Sprague-Dawley rats) and in vitro (human hepatocellular carcinoma - HepG2; colorectal adenocarcinoma cell lines - Caco-2) approaches. The first (healthy control) and second (control with chemically induced mammary carcinomas) group received saline only. The third, fourth and fifth group (all with breast cancer) were injected (i.p.) with a single dose of doxorubicin (8mg/kg), doxorubicin/fullerenol (100mg/kg of fullerenol 30min before administration of 8mg/kg doxorubicin) and fullerenol (100mg/kg), respectively. Two days after treatment, the rats were sacrificed. Results showed that treatment with doxorubicin alone caused significant changes in the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and alpha-hydroxybutyrate dehydrogenase (alpha-HBDH), as well as in the levels of malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px), total antioxidant status (TAS), glutathione reductase (GR), catalase (CAT) and superoxide dismutase (SOD) in the liver tissue. These effects were significantly reduced for all investigated parameters by pre-treatment with fullerenol but not for the MDA and GSH level. The HepG2 and Caco-2 cell lines were continuously treated with fullerenol for 12h, 24h, 48h and 96h at concentrations of 10microg/mL and 44microg/mL. With the aim of evaluating the modulating activity of fullerenol on doxorubicin-induced hepatotoxicity, the cell lines were simultaneously treated with doxorubicin (1microm; 5microm) and fullerenol (10microg/mL; 44microg/mL) in different combinations. When the cells are treated with 5microm doxorubicin along with the fullerenol, we can see a significant improvement of the cell capability during the entire time-line. We can conclude that fullerenol has cytotoxic effects on HepG2 by itself, but when the oxidative stress is too high the cytotoxic effects of fullerenol are overcome by its protective role as a strong antioxidant compound.
本研究旨在采用体内（雌性斯普拉格-道利大鼠）和体外（人肝癌细胞系-HepG2；结肠直肠腺癌细胞系-Caco-2）方法，研究富勒醇C60(OH)24对阿霉素诱导的肝毒性的潜在保护作用。第一组（健康对照组）和第二组（化学诱导乳腺癌对照组）仅接受生理盐水。第三、第四和第五组（均患有乳腺癌）分别腹腔注射单剂量阿霉素（8mg/kg）、阿霉素/富勒醇（在给予8mg/kg阿霉素前30分钟注射100mg/kg富勒醇）和富勒醇（100mg/kg）。治疗两天后，处死大鼠。结果表明，单独使用阿霉素治疗会导致血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、乳酸脱氢酶（LDH）和α-羟丁酸脱氢酶（α-HBDH）水平以及肝组织中丙二醛（MDA）、谷胱甘肽（GSH）、谷胱甘肽过氧化物酶（GSH-Px）、总抗氧化状态（TAS）、谷胱甘肽还原酶（GR）、过氧化氢酶（CAT）和超氧化物歧化酶（SOD）水平发生显著变化。通过富勒醇预处理，所有研究参数的这些影响均显著降低，但MDA和GSH水平除外。将HepG2和Caco-2细胞系分别用浓度为10μg/mL和44μg/mL的富勒醇连续处理12小时、24小时、48小时和96小时。为了评估富勒醇对阿霉素诱导的肝毒性的调节活性，将细胞系同时用阿霉素（1μM；5μM）和富勒醇（10μg/mL；44μg/mL）以不同组合进行处理。当细胞用5μM阿霉素和富勒醇一起处理时，在整个时间线内我们可以看到细胞能力有显著改善。我们可以得出结论，富勒醇本身对HepG2具有细胞毒性作用，但当氧化应激过高时，富勒醇作为一种强抗氧化化合物的保护作用会克服其细胞毒性作用。
