Effects of Senegenin against Hypoxia/Reoxygenation- Induced Injury in PC12 Cells

• Published in: Chinese journal of integrative medicine Vol. 22; no. 5; pp. 353 - 361
• Main Author: 朱小青 李学敏 赵严冬 纪惜銮 王彦平 付咏梅 王华东 陆大祥 戚仁斌
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.05.2016
• Subjects: Animals; Apoptosis - drug effects; Calcium - metabolism; Caspase 3 - metabolism; Caspase-3; Cell Hypoxia - drug effects; Cell Nucleus - drug effects; Cell Nucleus - metabolism; Drugs, Chinese Herbal - pharmacology; Flow Cytometry; Fluorescence; Intracellular Space - metabolism; Medicine; Medicine & Public Health; Membrane Potential, Mitochondrial - drug effects; NADPH Oxidases - metabolism; NADPH氧化酶; Neuroprotective Agents - pharmacology; Original Article; Oxygen - pharmacology; PC12 Cells; PC12细胞; Rats; Reactive Oxygen Species - metabolism; Staining and Labeling; 皂苷元; 细胞损伤; 缺氧/复氧损伤; 远志; senegenin; NAPDH oxidase; PC12 cells; reactive oxygen species; hypoxia/reoxygenation
• ISSN: 1672-0415; 1993-0402
• DOI: 10.1007/s11655-015-2091-8

Objective： To investigate the effect and the potential mechanism of Senegenin （Sen） against injury induced by hypoxia/reoxygenation （H/R） in highly differentiated PC12 cells. Methods： The cultured PC12 cells were treated with H/R in the presence or absence of Sen （60 μmol/L）. Four groups were included in the experiment： control group, H/R group, H/R＋Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase （LDH） in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential （△ψm）, reactive oxygen species （ROS） and intracellular free calcium （[Ca2＋]i） were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase （NOX） were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively. Results： Sen significantly elevated cell viability （P〈0.05）, decreased the leakage of LDH （P〈0.05） and apoptosis rate （P〈0.05） in H/R-injured PC12 cells. Sen maintained the value of △ψm （P〈0.05） and suppressed the activity of caspase-3 （P〈0.05）. Moreover, Sen reduced ROS accumulation （P〈0.05） and [Ca2＋]i increment （P〈0.05） by inhibiting the activity of NOX （P〈0.05）. Conclusion： Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca2＊]~, thereby suppressing the mitochondrial pathway of cellular apoptosis.