Zhao Lin, Yue Zengyaran, Wang Gang, Qin Jiahui, Ma Hongyue, Tang Decai, Yin Gang
School of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, PR China; School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, PR China.
School of Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, PR China.
Phytomedicine. 2025 Apr;139:156550. doi: 10.1016/j.phymed.2025.156550. Epub 2025 Feb 22.
Owing to its remarkable efficacy, cisplatin (CDDP) is widely used as a chemotherapeutic drug in clinical cancer treatment; however, its severe nephrotoxicity often leads to acute kidney injury (AKI), in turn adversely affecting patient treatment and quality of life. Smilax glabra Roxb. (TFL), a Chinese herbal medicine, has various pharmacological effects, including antitumour, anti-inflammatory, and antioxidant activities, with the antioxidant activity being of useful in the detoxification of heavy metal toxicity.
This study aimed to investigate, for the first time, the nephroprotective effects of TFL in alleviating CDDP-induced AKI and to elucidate its underlying mechanisms.
In vitro and in vivo models of AKI were established using CDDP induction. For the in vivo model, CDDP (20 mg/kg) was intraperitoneally injected on day 7 to induce AKI. TFL treatment was administered daily at doses of 1.95 and 3.9 g/kg starting from the day 1 and continuing for 10 consecutive days. Blood samples were collected on day 10 after 72-h of CDDP injection for analysis. Kidney pathology was observed using haematoxylin and eosin (HE) staining, and mitochondrial ultrastructure was assessed using transmission electron microscopy. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), HO-1, NQO1, caspase-3, and cytochrome C (CYT-C) were determined using western blotting, PCR, and immunofluorescence (IF). Adenosine triphosphate (ATP) levels, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) were measured using the corresponding kits. Lastly, reverse validation of the Nrf2/HO-1 pathway was performed using the Nrf2-specific inhibitor, ML385.
After induction with 40 μM CDDP, HK2 cells showed obvious mitochondrial damage, and the protein and mRNA expressions of Nrf2, HO-1, and NQO1 were inhibited, but gradually increased with TFL treatment. Furthermore, CDDP-induced AKI in mice was similar to the observations in the in vitro model using HK2 cells. The protective effects of TFL were reversed with ML385 therapy.
In both in vivo and in vitro experiments, TFL activated the Nrf2/HO-1 signalling pathway, promoting the expression of antioxidant enzymes and thereby ameliorating CDDP-induced oxidative stress, mitochondrial dysfunction and renal cell apoptosis.
顺铂（CDDP）因其显著的疗效，在临床癌症治疗中被广泛用作化疗药物；然而，其严重的肾毒性常导致急性肾损伤（AKI），进而对患者的治疗和生活质量产生不利影响。中药菝葜（TFL）具有多种药理作用，包括抗肿瘤、抗炎和抗氧化活性，其抗氧化活性有助于重金属毒性的解毒。
本研究旨在首次探讨TFL在减轻CDDP诱导的AKI中的肾保护作用，并阐明其潜在机制。
采用CDDP诱导建立AKI的体外和体内模型。对于体内模型，在第7天腹腔注射CDDP（20mg/kg）诱导AKI。从第1天开始，每天以1.95和3.9g/kg的剂量给予TFL治疗，并持续10天。在CDDP注射72小时后的第10天采集血样进行分析。使用苏木精和伊红（HE）染色观察肾脏病理，使用透射电子显微镜评估线粒体超微结构。使用蛋白质印迹法、PCR和免疫荧光（IF）测定核因子红细胞2相关因子2（Nrf2）、HO-1、NQO1、半胱天冬酶-3和细胞色素C（CYT-C）的表达水平。使用相应试剂盒测量三磷酸腺苷（ATP）水平、线粒体膜电位（MMP）和活性氧（ROS）。最后，使用Nrf2特异性抑制剂ML385对Nrf2/HO-1途径进行反向验证。
用40μM CDDP诱导后，HK2细胞表现出明显的线粒体损伤，Nrf2、HO-1和NQO1的蛋白质和mRNA表达受到抑制，但随着TFL治疗逐渐增加。此外，CDDP诱导的小鼠AKI与使用HK2细胞的体外模型中的观察结果相似。ML385治疗逆转了TFL的保护作用。
在体内和体外实验中，TFL均激活了Nrf2/HO-1信号通路，促进抗氧化酶的表达，从而改善CDDP诱导 的氧化应激、线粒体功能障碍和肾细胞凋亡。
