Fund programs: Health Research Project of Anhui Province （No. AHWJ2024Aa40016）； Occupational Medicine and Health Joint Research Project from Institute of Health and Medicine， Hefei Comprehensive National Science Center （No. OMH-2023-03）； Natural Science Foundation of Anhui Province （No. 2208085MH215）； Scientific Research Project of Inheritance and Innovation of Traditional Chinese Medicine in Anhui Province.（No. 2020cczd05）
Authors：Wang Jinhong1，2， Chen Tianyu1，2， Mao Lifang1，2， Zhao Yingjie1，2， Zhou Renpeng1，2， Hu Wei1，2， Lu Chao1，3
Keywords：genistein； Prdx6； chondrocytes； senescence； molecular docking； etoposide
DOI：10.19405/j.cnki.issn1000-1492.2026.04.007
〔Abstract〕 Objective To investigate the protective effect of genistein （Gen） on etoposide-induced chondrocyte senescence and its underlying mechanism.Methods The C28/I2 cell line was treated with different concentrations of Gen and etoposide， and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide， with Gen intervention. Senescence-associated β-galactosidase （SA-β-gal） staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 （Prdx6）， cyclin-dependent kinaseto clarify the functional necessity of Prdx6.Results Compared with the etoposide group， the C28/I2 chondrocyte viability significantly increased （P<0.01）， the expression ofsenescence-associated proteins p21 and p16 decreased （P<0.01， P<0.05）， the expression of senescence-associated genes p21 and p16 reduced （both P<0.01）， the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished （P<0.05， P<0.01）， and the proportion of SA-β-gal-positive cells decreased （P<0.01） in the Gen+etoposide group. Compared with the Control group， the expression of Prdx6 was downregulated in the etoposide group （P<0.05）. Compared with the etoposide group， the expression of Prdx6 was upregulated in the Gen+etoposide group （P<0.01）. Compared with the Control group， the GPx activity significantly decreased in the si-Prdx6 group （P<0.01）. Furthermore， compared with the si-Prdx6 group， the GPx activity increased in the si-Prdx6+Gen group （P<0.05）. Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown， the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group （both P<0.01）.Conclusion Gen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.