Liquid chromatography–mass spectrometry-based quantitative proteomics analysis reveals chondroprotective effects of astragaloside IV in interleukin-1β-induced SW1353 chondrocyte-like cells

Abstract Objective Chondrocyte apoptosis played a key role on the progression of Osteoarthritis (OA). Safe and effective drugs are urgently needed for the treatment of OA. Previous study reported that Astragaloside IV (ASG-IV) had exerted a protective effect against articular cartilage degeneration by promoting rapid proliferation of chondrocyte. Therefore, the aim of our study is to explore the effects and mechanisms of ASG-IV in chondrocyte apoptosis. Methods Isobaric Tags For Relative And Absolute Quantitation (iTRAQ)-based quantitative proteomics was used to quantitatively detect and map proteins in SW1353 chondrocyte-like cells pre-treated with ASG-IV or interleukin-1β (IL-1β) or ASG-IV + IL-1β. The iTRAQ-labeled peptides were fractionated by high-accuracy liquid chromatography–mass spectrometry (LC–MS). Cell apoptosis and differentially expressed proteins was detected by flow cytometry (FCM), quantitative real-time polymerase chain reaction (qRT-PCR), and western blotting, respectively. Results The apoptosis of the IL-1β-induced SW1353 cells treated with ASG-IV was greatly inhibited. Bioinformatics analysis revealed that gamma actin 1 ( ACTG1 ) and Yes Associated Protein 1 ( YAP1 ), participating in the Hippo signaling pathway and Vitronectin ( VTN ) and Collagen Type I Alpha 1 Chain ( COL1A1 ), involving in the extracellular matrix (ECM)-receptor interaction signaling pathway, were all significantly up-regulated in the IL-1β-induced SW1353 cells after treatment with ASG-IV. The qRT-PCR and Western blotting results confirmed the up-regulation of these four genes. Conclusion ASG-IV played a positive role in human osteoarthritic chondrocyte apoptosis, possibly through modulation of the Hippo signaling pathway by up-regulating YAP1 and ACTG1 expression, and also by up-regulating VTN and COL1A1 , which are involved in the ECM-receptor interaction pathway. Taken together, all the results suggested that ASG-IV had a novel therapeutic potential for the treatment of OA.