Department of Phisiology and Pharmacology, University Federal of Ceara, Fortaleza, Ceara, Brazil.
Department of Clinical and Toxicological Analysis, University Federal of Ceara, Fortaleza, Ceara, Brazil.
Eur J Pharmacol. 2019 Jul 15;855:1-9. doi: 10.1016/j.ejphar.2019.04.044. Epub 2019 Apr 29.
Acute Kidney Injury (AKI) is associated with high morbidity and mortality. Ischemia and reperfusion (I/R) are events that lead to AKI through hypoxia, reactive oxygen species (ROS) production, oxidative stress and apoptosis. We aimed to evaluate the mechanism of nephroprotection mediated by Bisabolol in human tubular kidney cells after injury by I/R in vitro. HK2 cells were exposed to I/R and treated with Bisabolol. Cell viability was accessed by MTT assay. Cells were submitted to flow cytometry to evaluate necrotic/apoptotic cells, reactive oxygen species production and mitochondrial transmembrane depolarization. TBARS and GSH were used as parameters of redox balance. Also, KIM-1 supernatant levels were measured. In order to identify an interaction between bisabolol and NOX4, molecular docking and enzymatic assays were performed. Expression of isoform NOX4 on treated cells was examined by western-blot. Finally, cells were visualized by scanning electron microscopy. Bisabolol improved cell viability and prevented cell death by apoptosis, indicated also by the decreased levels of KIM-1. It was observed a decrease on reactive oxygen species production and mitochondrial depolarization, with antioxidant regulation by increased GSH and decreased lipid peroxidation. It was also demonstrated that bisabolol treatment can inhibit NOX4. Finally, SEM images showed that bisabolol reduced I/R-induced cell damage. Bisabolol treatment protects HK2 cells against oxidative damage occasioned by I/R. This effect is related to inhibition of apoptosis, decrease on KIM-1 release, reactive oxygen species accumulation and mitochondrial dysfunction. Bisabolol inhibited NOX4 activity in the tubular cells, impairing reactive oxygen species synthesis.
急性肾损伤 (AKI) 与高发病率和死亡率相关。缺血再灌注 (I/R) 是通过缺氧、活性氧 (ROS) 产生、氧化应激和细胞凋亡导致 AKI 的事件。我们旨在评估毕沙罗醇在体外 I/R 损伤后对人肾小管肾细胞的保护机制。将 HK2 细胞暴露于 I/R 并用毕沙罗醇处理。通过 MTT 测定法评估细胞活力。通过流式细胞术评估坏死/凋亡细胞、活性氧产生和线粒体跨膜去极化。TBARS 和 GSH 用于评估氧化还原平衡的参数。还测量了 KIM-1 上清液水平。为了鉴定毕沙罗醇和 NOX4 之间的相互作用，进行了分子对接和酶测定。通过 Western blot 检查处理细胞中 NOX4 同工型的表达。最后，通过扫描电子显微镜观察细胞。毕沙罗醇通过减少 KIM-1 水平来改善细胞活力并防止细胞凋亡。还观察到活性氧产生和线粒体去极化减少，通过增加 GSH 和减少脂质过氧化来调节抗氧化作用。还表明，毕沙罗醇处理可以抑制 NOX4。最后，SEM 图像显示，毕沙罗醇可减少 I/R 诱导的细胞损伤。毕沙罗醇治疗可保护 HK2 细胞免受 I/R 引起的氧化损伤。这种作用与抑制细胞凋亡、减少 KIM-1 释放、活性氧积累和线粒体功能障碍有关。毕沙罗醇抑制肾小管细胞中 NOX4 的活性，从而抑制活性氧的合成。
