Tan Shiqing, Bai Jie, Xu Mingxi, Zhang Longying, Wang Ying
The Second Affiliated Hospital, Dalian Medical University, Dalian, China.
Nutrition and Food Hygiene, Dalian Medical University, Dalian, China.
Front Pharmacol. 2022 Apr 13;13:841330. doi: 10.3389/fphar.2022.841330. eCollection 2022.
Doxorubicin (DOX) has been widely used in cancer treatment. However, DOX can cause a range of significant side effects, of which hepatotoxicity is a common one, and therefore limits its clinical use. Pterostilbene (PTS) has been shown to exhibit anti-oxidant and anti-inflammatory effects in the treatment of liver diseases but whether PTS could protect against hepatotoxicity in DOX-treated mice is unknown. In our study, we use C57/BL6J mice and the HepG2 cell line. We divided the mice in 4 groups: the control, the PTS treatment, the DOX treatment, and the DOX + PTS treatment group. Liver histopathology was judged by performing hematoxylin-eosin and Masson staining. Immunohistochemistry was used to perform the expression of NLRP3. The levels of serum alanine transaminase (ALT) and aspartate transaminase (AST) were evaluated. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and DCFH-DA staining were used to evaluate the oxidative injury. Western blot and real-time PCR were applied to evaluate the expressions of proteins and mRNA. MTT was used to evaluate DOX-induced cell injury and the protective effects of PTS. Recombinant Trx-1 was used to analyze the mechanism of PTS. A TUNEL assay was used to detect apoptosis in DOX-induced HepG2 cells and the protective effects of PTS. PTS ameliorated DOX-induced liver pathological changes and the levels of AST and ALT. PTS also decreased the level of MDA, increased the level of SOD, GSH, and the expression of Trx-1 in DOX-treated mice. PTS decreased the levels of NLRP3 and IL-1β mRNA and the expressions of their proteins in DOX-treated mice. In addition, PTS also decreased the expression of Cleaved Caspase-3 and BAX and increased the expression of BCL-2. , after treatment with recombinant Trx-1, ROS and NLRP3 inflammasome were both decreased. Treatment with PTS could rescue the downregulation of Trx-1, decreased the ROS level and the NLRP3 inflammasome, and protected HepG2 cells against DOX-induced apoptosis. The results show that PTS exhibits protective effects against DOX-induced liver injuries suppression of oxidative stress, fibrosis, NLRP3 inflammasome stimulation, and cell apoptosis which might lead to a new approach of preventing DOX-induced hepatotoxicity.
阿霉素（DOX）已广泛应用于癌症治疗。然而，DOX会引起一系列严重的副作用，其中肝毒性较为常见，因此限制了其临床应用。已表明紫檀芪（PTS）在肝病治疗中具有抗氧化和抗炎作用，但PTS是否能预防DOX处理小鼠的肝毒性尚不清楚。在我们的研究中，我们使用C57/BL6J小鼠和HepG2细胞系。我们将小鼠分为4组：对照组、PTS处理组、DOX处理组和DOX + PTS处理组。通过苏木精-伊红染色和Masson染色判断肝脏组织病理学。免疫组织化学用于检测NLRP3的表达。评估血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。使用丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）水平以及DCFH-DA染色评估氧化损伤。应用蛋白质印迹法和实时定量PCR评估蛋白质和mRNA的表达。MTT用于评估DOX诱导的细胞损伤以及PTS的保护作用。使用重组Trx-1分析PTS的作用机制。TUNEL法用于检测DOX诱导的HepG2细胞凋亡以及PTS的保护作用。PTS改善了DOX诱导的肝脏病理变化以及AST和ALT水平。PTS还降低了DOX处理小鼠的MDA水平，提高了SOD、GSH水平以及Trx-1的表达。PTS降低了DOX处理小鼠中NLRP3和IL-1β mRNA水平及其蛋白质表达。此外，PTS还降低了裂解的半胱天冬酶-3和BAX的表达，并增加了BCL-2的表达。用重组Trx-1处理后，活性氧（ROS）和NLRP3炎性小体均减少。PTS处理可挽救Trx-1的下调，降低ROS水平和NLRP3炎性小体，并保护HepG2细胞免受DOX诱导的凋亡。结果表明，PTS对DOX诱导的肝损伤具有保护作用，可抑制氧化应激、纤维化、NLRP3炎性小体激活和细胞凋亡，这可能为预防DOX诱导的肝毒性提供一种新方法。
