Y Zhu1, T Wang1,2, N Dai3, M Deng1, H Liu1, X Tong1, L Li1. 1. Department of Pharmacology, College of Integrated Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China. 2. Anhui Provincial Key Laboratory of Traditional Chinese Medicine Compounds, Hefei 230012, China. 3. Department of Andrology, First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230601, China.
Abstract
OBJECTIVE: To study the protective effect of hyperoside (Hyp) against ydrogen peroxide (H2O2)- induced oxidative damage in mouse spermatocytes GC-2 cells and explore the role of the Keap1/Nrf2/HO-1 pathway in this protective mechanism. METHODS: GC-2 cells were treated with 2.5 mmol/L azaacetylcysteine (NAC), 50, 100, and 200 μmol/L hyperoside, or the culture medium for 48 h before exposure to H2O2 (150 μmol/L) for 2 h. CCK-8 assay was used to detect the changes in cell viability, and cell apoptosis was analyzed using flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) activity and malondialdehyde (MDA) in the culture medium. Western blotting and RT-qPCR were used to detect the protein and mRNA expression levels of nuclear factor erythroid 2-related factor2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), and heme oxygenase-1 (HO-1); the nuclear translocation of Nrf2 was detected using immunofluorescence assay. RESULTS: Exposure to H2O2 significantly lowered the proliferation rate, reduced the activities of SOD, GSH and CAT, and obviously increased MDA content, cell apoptosis rate, and the expressions of Keap1 and Nrf2 mRNA and Keap1 protein in GC-2 cells (P < 0.05 or 0.01). Treatment of the cells prior to H2O2 exposure with either NAC or 200 μmol/L hyperoside significantly increased the cell proliferation rate, enhanced the activities of SOD, GSH-PX and CAT, and lowered MDA content and cell apoptosis rate (P < 0.05). Treatment with 200 μmol/L hyperoside significantly decreased the mRNA and protein expressions of Keap1 and increased the expressions of HO-1 mRNA and the protein expressions of Nrf2 and HO-1 (P < 0.05 or 0.01). Hyperoside also caused obvious nuclear translocation of Nrf2 in the cells (P < 0.05). CONCLUSION: Hyperoside protects GC-2 cells against H2O2- induced oxidative damage possibly by activation of the Keap1/Nrf2/HO-1 signaling pathway.
OBJECTIVE: To study the protective effect of hyperoside (Hyp) against ydrogen peroxide (H2O2)- induced oxidative damage in mouse spermatocytes GC-2 cells and explore the role of the Keap1/Nrf2/HO-1 pathway in this protective mechanism. METHODS: GC-2 cells were treated with 2.5 mmol/L azaacetylcysteine (NAC), 50, 100, and 200 μmol/L hyperoside, or the culture medium for 48 h before exposure to H2O2 (150 μmol/L) for 2 h. CCK-8 assay was used to detect the changes in cell viability, and cell apoptosis was analyzed using flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) activity and malondialdehyde (MDA) in the culture medium. Western blotting and RT-qPCR were used to detect the protein and mRNA expression levels of nuclear factor erythroid 2-related factor2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), and heme oxygenase-1 (HO-1); the nuclear translocation of Nrf2 was detected using immunofluorescence assay. RESULTS: Exposure to H2O2 significantly lowered the proliferation rate, reduced the activities of SOD, GSH and CAT, and obviously increased MDA content, cell apoptosis rate, and the expressions of Keap1 and Nrf2 mRNA and Keap1 protein in GC-2 cells (P < 0.05 or 0.01). Treatment of the cells prior to H2O2 exposure with either NAC or 200 μmol/L hyperoside significantly increased the cell proliferation rate, enhanced the activities of SOD, GSH-PX and CAT, and lowered MDA content and cell apoptosis rate (P < 0.05). Treatment with 200 μmol/L hyperoside significantly decreased the mRNA and protein expressions of Keap1 and increased the expressions of HO-1 mRNA and the protein expressions of Nrf2 and HO-1 (P < 0.05 or 0.01). Hyperoside also caused obvious nuclear translocation of Nrf2 in the cells (P < 0.05). CONCLUSION: Hyperoside protects GC-2 cells against H2O2- induced oxidative damage possibly by activation of the Keap1/Nrf2/HO-1 signaling pathway.
Authors: Mafalda V Moreira; Sara C Pereira; Bárbara Guerra-Carvalho; David F Carrageta; Soraia Pinto; Alberto Barros; Branca M Silva; Pedro F Oliveira; Marco G Alves Journal: Cell Physiol Biochem Date: 2022-01-08
Authors: Ashok Agarwal; Neel Parekh; Manesh Kumar Panner Selvam; Ralf Henkel; Rupin Shah; Sheryl T Homa; Ranjith Ramasamy; Edmund Ko; Kelton Tremellen; Sandro Esteves; Ahmad Majzoub; Juan G Alvarez; David K Gardner; Channa N Jayasena; Jonathan W Ramsay; Chak Lam Cho; Ramadan Saleh; Denny Sakkas; James M Hotaling; Scott D Lundy; Sarah Vij; Joel Marmar; Jaime Gosalvez; Edmund Sabanegh; Hyun Jun Park; Armand Zini; Parviz Kavoussi; Sava Micic; Ryan Smith; Gian Maria Busetto; Mustafa Emre Bakırcıoğlu; Gerhard Haidl; Giancarlo Balercia; Nicolás Garrido Puchalt; Moncef Ben-Khalifa; Nicholas Tadros; Jackson Kirkman-Browne; Sergey Moskovtsev; Xuefeng Huang; Edson Borges; Daniel Franken; Natan Bar-Chama; Yoshiharu Morimoto; Kazuhisa Tomita; Vasan Satya Srini; Willem Ombelet; Elisabetta Baldi; Monica Muratori; Yasushi Yumura; Sandro La Vignera; Raghavender Kosgi; Marlon P Martinez; Donald P Evenson; Daniel Suslik Zylbersztejn; Matheus Roque; Marcello Cocuzza; Marcelo Vieira; Assaf Ben-Meir; Raoul Orvieto; Eliahu Levitas; Amir Wiser; Mohamed Arafa; Vineet Malhotra; Sijo Joseph Parekattil; Haitham Elbardisi; Luiz Carvalho; Rima Dada; Christophe Sifer; Pankaj Talwar; Ahmet Gudeloglu; Ahmed M A Mahmoud; Khaled Terras; Chadi Yazbeck; Bojanic Nebojsa; Damayanthi Durairajanayagam; Ajina Mounir; Linda G Kahn; Saradha Baskaran; Rishma Dhillon Pai; Donatella Paoli; Kristian Leisegang; Mohamed Reza Moein; Sonia Malik; Onder Yaman; Luna Samanta; Fouad Bayane; Sunil K Jindal; Muammer Kendirci; Baris Altay; Dragoljub Perovic; Avi Harlev Journal: World J Mens Health Date: 2019-05-28 Impact factor: 5.400