Protective effect of pentoxifylline on human umbilical vein endothelial cell injury induced by tert-butyl hydroperoxide
Journal Title: Journal of Air Force Medical University - Year 2023, Vol 44, Issue 3
Abstract
Objective To investigate the protective effect of pentoxifylline (PTX) on oxidative stress and mitochondrial function in human umbilical vein endothelial cells (HUVECs) induced by tert-butyl hydroperoxide (t-BHP). Methods t-BHP was used to induce HUVECs injury model, and PTX was pretreated to screen suitable intervention dose. The changes of mitochondrial mass, mitochondrial reactive oxygen species (ROS), and mitochondrial membrane potential were detected by Mito Tracker Green, Mito SOX, and Rho 123 probes, respectively. Then the cells were divided into control group, t-BHP group, PTX pretreatment group and simple PTX group. The control group did not receive any treatment, t-BHP group was treated with 105 μmol / L t-BHP, simple PTX group with 1 mg / L PTX, and PTX pretreatment group with 105 μmol / L t-BHP and 1 mg / L PTX. MTT assay was used to detect cell viability. The levels of lactic dehydrogenase ( LDH), malondialdehyde (MDA), oxidized glutathione (GSSG), reduced glutathione (GSH), adenosine triphosphate (ATP) and the activity of superoxide dismutase (SOD) were detected by test kits. Western blotting was used to detect the expression levels of SOD1, SOD2 and nuclear factor erythroid 2-related factor 2 ( Nrf-2). The activities of Na + , K + -ATPase and Ca2 + , Mg2 + -ATPase were determined by phosphate method. Results Horns method calculated and measured the median lethal dose of HUVECs exposed to t-BHP for 6 h was 105 μmol / L. When PTX was pretreated for 12 h and the concentration was more than 1 mg / L, the cell viability of HUVECs was increased ( P < 0. 05). Compared with the control group, the green fluorescence intensity of Mito Tracker Green and Rho 123 decreased, while the red fluorescence intensity of Mito SOX increased after t-BHP treatment. Compared with t-BHP group, PTX pretreatment group increased cell viability, decreased the levels of LDH, MDA and GSSG in cell culture supernatant, SOD activity and Nrf-2 protein expression in HUVECs (P < 0. 05), and increased levels of GSH, GSH / GSSG ratio, SOD1 protein expression, ATP, and the activities of Na + , K + -ATPase and Ca2 + , Mg2 + -ATPase (P < 0. 05). Conclusion PTX can protect HUVECs from t-BHP-induced oxidative stress and mitochondrial damage.
Authors and Affiliations
LIU Mengmeng, ZHAO Chenqian, GUO Peng, LIU Jiangzheng, KONG Deqin, REN Xiaoting, LIU Rui,HAI Chunxu, ZHANG Xiaodi
Keywords
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