School of Medicine, Shanghai University, Shanghai 200444, China; Shanghai Health Commission Key Lab of Artificial Intelligence-Based Management of Inflammation and Chronic Diseases, Shanghai Pudong Gongli Hospital, Secondary Military Medical University, Shanghai 200135, China.
Shanghai Health Commission Key Lab of Artificial Intelligence-Based Management of Inflammation and Chronic Diseases, Shanghai Pudong Gongli Hospital, Secondary Military Medical University, Shanghai 200135, China; School of Basic Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China.
J Integr Med. 2024 May;22(3):286-294. doi: 10.1016/j.joim.2024.03.007. Epub 2024 Mar 16.
Research has shown that celastrol can effectively treat a variety of diseases, yet when passing a certain dosage threshold, celastrol becomes toxic, causing complications such as liver and kidney damage and erythrocytopenia, among others. With this dichotomy in mind, it is extremely important to find ways to preserve celastrol's efficacy while reducing or preventing its toxicity.
In this study, insulin-resistant HepG2 (IR-HepG2) cells were prepared using palmitic acid and used for in vitro experiments. IR-HepG2 cells were treated with celastrol alone or in combination with N-acetylcysteine (NAC) or ferrostatin-1 (Fer-1) for 12, 24 or 48 h, at a range of doses. Cell counting kit-8 assay, Western blotting, quantitative reverse transcription-polymerase chain reaction, glucose consumption assessment, and flow cytometry were performed to measure celastrol's cytotoxicity and whether the cell death was linked to ferroptosis.
Celastrol treatment increased lipid oxidation and decreased expression of anti-ferroptosis proteins in IR-HepG2 cells. Celastrol downregulated glutathione peroxidase 4 (GPX4) mRNA. Molecular docking models predicted that solute carrier family 7 member 11 (SLC7A11) and GPX4 were covalently bound by celastrol. Importantly, we found for the first time that the application of ferroptosis inhibitors (especially NAC) was able to reduce celastrol's toxicity while preserving its ability to improve insulin sensitivity in IR-HepG2 cells.
One potential mechanism of celastrol's cytotoxicity is the induction of ferroptosis, which can be alleviated by treatment with ferroptosis inhibitors. These findings provide a new strategy to block celastrol's toxicity while preserving its therapeutic effects. Please cite this article as: Liu JJ, Zhang X, Qi MM, Chi YB, Cai BL, Peng B, Zhang DH. Ferroptosis inhibitors reduce celastrol toxicity and preserve its insulin sensitizing effects in insulin resistant HepG2 cells. J Integr Med. 2024; 22(3): 286-294.
研究表明，雷公藤红素可有效治疗多种疾病，但当其剂量超过一定阈值时，会产生毒性，导致肝、肾损伤和红细胞减少等并发症。鉴于这种双重性，寻找既能保持雷公藤红素疗效又能降低或预防其毒性的方法非常重要。
本研究采用棕榈酸构建胰岛素抵抗 HepG2（IR-HepG2）细胞模型，进行体外实验。用不同浓度的雷公藤红素单独或联合 N-乙酰半胱氨酸（NAC）或 ferrostatin-1（Fer-1）处理 IR-HepG2 细胞 12、24 或 48 h，采用细胞计数试剂盒-8 检测、Western blot、实时定量聚合酶链反应、葡萄糖消耗评估和流式细胞术检测雷公藤红素的细胞毒性及细胞死亡是否与铁死亡有关。
雷公藤红素处理可增加 IR-HepG2 细胞的脂质氧化，降低抗铁死亡蛋白的表达。雷公藤红素下调谷胱甘肽过氧化物酶 4（GPX4）mRNA 的表达。分子对接模型预测溶质载体家族 7 成员 11（SLC7A11）和 GPX4 与雷公藤红素发生共价结合。重要的是，我们首次发现应用铁死亡抑制剂（尤其是 NAC）可降低雷公藤红素的毒性，同时保持其改善 IR-HepG2 细胞胰岛素敏感性的作用。
雷公藤红素的细胞毒性的一个潜在机制是诱导铁死亡，铁死亡抑制剂的治疗可缓解雷公藤红素的毒性。这些发现为阻断雷公藤红素的毒性而保留其治疗效果提供了新策略。请引用本文为：刘静静，张霞，齐苗苗，迟艳彬，蔡兵林，彭波，张大华。铁死亡抑制剂降低雷公藤红素在胰岛素抵抗 HepG2 细胞中的毒性并保留其胰岛素增敏作用。 解放军医学杂志。2024; 22(3): 286-294.
