School of Biological Sciences, University of Northern Colorado, Greeley, CO 80639, USA.
School of Sport and Exercise Science, University of Northern Colorado, Greeley, CO 80639, USA.
Nutrients. 2020 Dec 24;13(1):41. doi: 10.3390/nu13010041.
: Treatment with the chemotherapy drug doxorubicin (DOX) may lead to toxicities that affect non-cancer cells including the liver. Supplementing the diet with creatine (Cr) has been suggested as a potential intervention to minimize DOX-induced side effects, but its effect in alleviating DOX-induced hepatoxicity is currently unknown. Therefore, we aimed to examine the effects of Cr supplementation on DOX-induced liver damage. Male Sprague-Dawley rats were fed a diet supplemented with 2% Cr for four weeks, 4% Cr for one week followed by 2% Cr for three more weeks, or control diet for four weeks. Animals then received either a bolus i.p. injection of DOX (15 mg/kg) or saline as a placebo. Animals were then sacrificed five days-post injection and markers of hepatoxicity were analyzed using the liver-to-body weight ratio, aspartate transaminase (AST)-to- alanine aminotransferase (ALT) ratio, alkaline phosphatase (ALP), lipemia, and T-Bilirubin. In addition, hematoxylin and eosin (H&E) staining, Picro-Sirius Red staining, and immunofluorescence staining for CD45, 8-OHdG, and β-galactosidase were performed to evaluate liver morphology, fibrosis, inflammation, oxidative stress, and cellular senescence, respectively. The mRNA levels for biomarkers of liver fibrosis, inflammation, oxidative stress, and senescence-related genes were measured in liver tissues. Chromosomal stability was evaluated using global DNA methylation ELISA. The ALT/AST ratio and liver to body weight ratio tended to increase in the DOX group, and Cr supplementation tended to attenuate this increase. Furthermore, elevated levels of liver fibrosis, inflammation, oxidative stress, and senescence were observed with DOX treatment, and Cr supplementation prior to DOX treatment ameliorated this hepatoxicity. Moreover, DOX treatment resulted in chromosomal instability (i.e., altered DNA methylation profile), and Cr supplementation showed a tendency to restore chromosomal stability with DOX treatment. The data suggest that Cr protected against DOX-induced hepatotoxicity by attenuating fibrosis, inflammation, oxidative stress, and senescence.
：多柔比星（DOX）化疗药物治疗可能导致影响非癌细胞的毒性，包括肝脏。补充肌酸（Cr）已被建议作为减轻 DOX 诱导的副作用的潜在干预措施，但减轻 DOX 诱导的肝毒性的效果尚不清楚。因此，我们旨在研究 Cr 补充对 DOX 诱导的肝损伤的影响。雄性 Sprague-Dawley 大鼠用补充 2% Cr 的饮食喂养四周，4% Cr 喂养一周，然后再用 2% Cr 喂养三周，或用补充 4 周对照饮食。然后，动物接受腹腔内单次 DOX（15mg/kg）注射或生理盐水作为安慰剂。动物在注射后五天处死，通过肝重/体重比、天冬氨酸转氨酶（AST）/丙氨酸转氨酶（ALT）比、碱性磷酸酶（ALP）、脂血症和 T-胆红素分析肝毒性标志物。此外，进行苏木精和伊红（H&E）染色、苦味酸-天狼星红染色和 CD45、8-OHdG 和β-半乳糖苷酶的免疫荧光染色，分别评估肝脏形态、纤维化、炎症、氧化应激和细胞衰老。用肝脏组织中肝纤维化、炎症、氧化应激和衰老相关基因的生物标志物的 mRNA 水平进行测量。用全基因组 DNA 甲基化 ELISA 评估染色体稳定性。DOX 组的 ALT/AST 比和肝重/体重比趋于增加，Cr 补充趋于减弱这种增加。此外，用 DOX 处理观察到肝纤维化、炎症、氧化应激和衰老增加，Cr 补充在 DOX 处理前可改善这种肝毒性。此外，DOX 处理导致染色体不稳定（即改变的 DNA 甲基化谱），Cr 补充在 DOX 处理时显示出恢复染色体稳定性的趋势。数据表明，Cr 通过减轻纤维化、炎症、氧化应激和衰老来保护免受 DOX 诱导的肝毒性。
