Bakhshii Saba, Khezri Saleh, Ahangari Roya, Jahedsani Asal, Salimi Ahmad
Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran.
Students Research Committee, Faculty of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran.
Drug Dev Res. 2021 Sep;82(6):835-843. doi: 10.1002/ddr.21790. Epub 2021 Jan 13.
Clozapine (CLZ) is unusually efficient in psychotic diseases. Nonetheless, its use is confined due to potentially life-threatening adverse events, including cardiotoxicity. Since the cardiotoxicity of CLZ is mediated through the generation of active metabolites, free radical, and inflammation. Here, we tested this hypothesis that kaempferol (KP) as antioxidant and anti-inflammatory agent could attenuate CLZ-induced mitochondrial/lysosomal and oxidative damages in rat ventricular cardiomyocytes. Rat ventricular cardiomyocytes were isolated by collagenase perfusion. Then isolated cardiomyocytes were simultaneously treated with different concentrations of KP (10, 20, and 50 μM) and CLZ (50 μM) for 4 h at 37°C. After 4 h of incubation, using by flow cytometry and biochemical evaluations, the parameters of cellular toxicity including: cell viability, reactive oxygen species (ROS) formation, mitochondria membrane potential (ΔΨm) collapse, lysosomal membrane integrity, malondialdehyde, and oxidized/reduced glutathione were analyzed. The results showed that CLZ (50 μM) induced a significant increase in cytotoxicity, ROS formation, mitochondrial membrane potential collapse, lipid peroxidation, and oxidative stress while KP reverted the above toxic effect of CLZ on isolated cardiomyocytes. Our data suggest that KP prevents and reverses CLZ-induced oxidative and mitochondrial/lysosomal damages in isolated cardiomyocytes, providing an experimental basis for clinical treatment on CLZ-induced cardiotoxicity.
氯氮平（CLZ）在治疗精神疾病方面异常有效。然而，由于其潜在的危及生命的不良事件，包括心脏毒性，其应用受到限制。由于CLZ的心脏毒性是通过活性代谢产物、自由基的产生和炎症介导的。在此，我们验证了这一假设，即作为抗氧化剂和抗炎剂的山奈酚（KP）可以减轻CLZ诱导的大鼠心室心肌细胞的线粒体/溶酶体损伤和氧化损伤。通过胶原酶灌注分离大鼠心室心肌细胞。然后将分离的心肌细胞在37℃下同时用不同浓度的KP（10、20和50μM）和CLZ（50μM）处理4小时。孵育4小时后，通过流式细胞术和生化评估，分析细胞毒性参数，包括：细胞活力、活性氧（ROS）形成、线粒体膜电位（ΔΨm）崩溃、溶酶体膜完整性、丙二醛以及氧化型/还原型谷胱甘肽。结果表明，CLZ（50μM）诱导细胞毒性、ROS形成、线粒体膜电位崩溃、脂质过氧化和氧化应激显著增加，而KP可逆转CLZ对分离心肌细胞的上述毒性作用。我们的数据表明，KP可预防和逆转CLZ诱导的分离心肌细胞的氧化损伤以及线粒体/溶酶体损伤，为CLZ诱导的心脏毒性的临床治疗提供了实验依据。
