Cuscuta chinensis seeds water extraction protecting murine osteoblastic MC3T3-E1 cells against tertiary butyl hydroperoxide induced injury |
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Authors: | Jian-mei Gao Ran Li Lei Zhang Li-long Jia Xi-xiang Ying De-qiang Dou Jian-chun Li Hai-bo Li |
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Institution: | 1. Department of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China;2. Department of Medical Science, Shandong Wanjie Medical University, 246 Xiwaihuan Road, Zibo 255213, China;3. School of Basic Medical Sciences, Liaoning University of TCM, 79 Chongshan East Road, Shenyang 110032, China;4. School of Pharmacy, Liaoning University of TCM, 1 Shengming Road, Dalian 116600, China |
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Abstract: | Ethnopharmacological relevanceCuscuta chinensis (C. chinensis) is a well-known traditional Chinese herb that has been used to treat heart disease, diabetes, liver injury, cancer, and aging.Materials and methodsMurine osteoblastic MC3T3-E1 cells were treated with various concentrations of C. chinensis water extraction at different time intervals. The antioxidant effect of C. chinensis on MC3T3-E1 cells was evaluated using MTT and TUNEL assays. The effect of C. chinensis on cell cycle was analyzed by flow cytometry with propidium iodide. Lipid peroxidation was measured by the HPLC method. The cellular redox status was determined from the reduced glutathione to oxidized glutathione ratio (GSH/GSSG) and the enzymes involved in glutathione metabolism, including glutathione reductase (GR), Glutathione S-transferase (GST), and Glucose-6-phosphate dehydrogenase (G6PD). The changes in relative mitochondrial transmembrane potential (ΔΨm) in the MC3T3-E1 cells were analyzed with rhodamine 123 staining. Western blot analysis was used to evaluate the levels of cytochrome c (cyto c), Bax, Bcl-2, caspase 3, Sirt3, and IDH2 expressions.ResultsThe C. chinensis water extraction protects tertiary butyl hydroperoxide (TBHP)-treated MC3T3-E1 cells from death in a dose-dependent manner. C. chinensis treatment significantly inhibited the reactive oxygen species (ROS) generation, malondialdehyde (MDA) production, and increased the activity of superoxide dismutase (SOD), GR, GST, and G6PD. The release of cyto c from mitochondria was reduced by C. chinensis, which increased the expression of antiapoptotic IDH2, Sirt3, and Bcl-2 and decreased the expression of Bax, cyto c, and caspase 3.ConclusionsC. chinensis modulated the oxidative stress-induced apoptosis in MC3T3-E1 cells, probably due to its antioxidant activity and functioning via mitochondria-dependent pathways. |
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Keywords: | Cuscuta chinensis Water extraction MC3T3-E1 Reactive oxygen species Tertiary butyl hydroperoxide |
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