Huali Luo1, Ling Yao2, Yudi Zhang3, Rongheng Li4. 1. Department of Integrated Traditional Chinese and Western Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China; Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, Chongqing Medical University, Chongqing 400016, China. 2. Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, Chongqing Medical University, Chongqing 400016, China. 3. Department of Integrated Traditional Chinese and Western Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China. 4. Department of Integrated Traditional Chinese and Western Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China. Electronic address: Lrongheng1231@163.com.
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 YAP1and 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.
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 YAP1and 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.