雷公藤甲素是从传统中药雷公藤（T. wilfordii）Hook F中分离出来的主要生物活性和毒性成分。它具有有效的抗肿瘤、免疫抑制和抗炎等生物活性。然而，其严重的全身毒性阻碍了其临床应用。临床上常用的雷公藤制剂有雷公藤苷片和雷公藤甲素片两种。然而，它们的不良副作用，特别是肝毒性，限制了它们的安全使用。因此，寻找雷公藤甲素强效、特异性的解毒药物至关重要。本研究旨在探讨螺内酯对雷公藤甲素所致肝毒性的解毒作用及潜在作用机制，为雷公藤甲素提供潜在的解毒策略，从而促进雷公藤甲素的安全应用。临床环境中的 T. wilfordii 制剂。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑 (MTT) 测定和结晶紫染色评估细胞活力。使用 4',6-二脒基-2-苯基吲哚 (DAPI) 染色观察核碎片，并通过蛋白质印迹分析蛋白质表达。通过检查螺内酯对血清丙氨酸氨基转移酶和天冬氨酸氨基转移酶水平的影响，以及雷公藤甲素诱导的急性肝毒性小鼠模型的肝脏病理学，评估螺内酯对雷公藤甲素诱导的肝毒性的抑制作用。此外，还进行了生存测定，以研究螺内酯对暴露于致死剂量的雷公藤甲素的小鼠的生存率的影响。通过 5-乙炔基尿苷染色评估螺内酯对雷公藤内酯诱导的整体转录抑制的影响。雷公藤内酯处理降低了肝癌细胞和肝细胞中的细胞活力，增加了核碎片和裂解的 caspase-3 水平。它还增加了丙氨酸转氨酶和天冬氨酸转氨酶的水平，导致肝细胞肿胀和坏死，并导致11只小鼠中7只死亡。通过螺内酯预处理可以减轻上述影响。此外，分子机制探索揭示螺内酯抑制雷公藤甲素诱导的DNA引导的RNA聚合酶II亚基RPB1降解，从而增加新生RNA的5-乙炔基尿苷染色的荧光强度。本研究表明螺内酯对雷公藤甲素的肝毒性表现出有效的解毒作用，通过抑制雷公藤甲素诱导的 RPB1 降解，进而延缓受影响细胞中的整体转录抑制。这些发现提出了雷公藤甲素的潜在解毒策略，可能有助于安全使用雷公藤制剂。版权所有 © 2024。由 Elsevier B.V. 出版。

Triptolide is a major bioactive and toxic ingredient isolated from the traditional Chinese herb Tripterygium wilfordii (T. wilfordii) Hook F. It exhibits potent antitumor, immunosuppressive, and anti-inflammatory biological activities; however, its clinical application is hindered by severe systemic toxicity. Two preparations of T. wilfordii, including T. wilfordii glycoside tablets and T. wilfordii tablets, containing triptolide, are commonly used in clinical practice. However, their adverse side effects, particularly hepatotoxicity, limit their safe use. Therefore, it is crucial to discover potent and specific detoxification medicines for triptolide.This study aimed to investigate the detoxification effects and potential mechanism of action of spironolactone on triptolide-induced hepatotoxicity to provide a potential detoxifying strategy for triptolide, thereby promoting the safe applications of T. wilfordii preparations in clinical settings.Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and crystal violet staining. Nuclear fragmentation was visualized using 4',6-diamidino-2-phenylindole (DAPI) staining, and protein expression was analyzed by Western blotting. The inhibitory effect of spironolactone on triptolide-induced hepatotoxicity was evaluated by examining the effects of spironolactone on serum alanine aminotransferase and aspartate aminotransferase levels, as well as liver pathology in a mouse model of triptolide-induced acute hepatotoxicity. Furthermore, a survival assay was performed to investigate the effects of spironolactone on the survival rate of mice exposed to a lethal dose of triptolide. The effect of spironolactone on triptolide-induced global transcriptional repression was assessed through 5-ethynyl uridine staining.Triptolide treatment decreased the cell viability, increased the nuclear fragmentation and the cleaved caspase-3 levels in both hepatoma cells and hepatocytes. It also increased the alanine aminotransferase and aspartate aminotransferase levels, induced the hepatocyte swelling and necrosis, and led to seven deaths out of 11 mice. The above effects could be mitigated by pretreatment with spironolactone. Additionally, molecular mechanism exploration unveiled that spironolactone inhibited triptolide-induced DNA-directed RNA polymerase II subunit RPB1 degradation, consequently increased the fluorescence intensity of 5-ethynyl uridine staining for nascent RNA.This study shows that spironolactone exhibits a potent detoxification role against triptolide hepatotoxicity, through inhibition of RPB1 degradation induced by triptolide and, in turn, retardation of global transcriptional inhibition in affected cells. These findings suggest a potential detoxification strategy for triptolide that may contribute to the safe use of T. wilfordii preparations.Copyright © 2024. Published by Elsevier B.V.