Influence of 1,25(OH)2D3 on proteoglycan 4 gene expression and secretion by rabbit synovial membrane
WANG Zhiqiang1, GUANG Chaoling2, ZHU Hui3, LI Xianhui2
1. Special Medical Center of Chinese People’s Armed Police Force,Tianjin 300162,China;
2. Department of Health Service,Logistics University of People’s Armed Police Force,Tianjin,300309,China;
3. Foundation depontment,Logistics University of People’s Armed Police Force,Tianjin,300309,China
Objective To investigate the effects of 1,25(OH)2D3 (the active form of vitamin D) on proteoglycan 4 (PRG4) gene expression and secretion by rabbit synovial membrane in order to find an effective scheme for protecting synovial joints and preventing osteoarthritis.Methods The synovial membrane from New Zealand rabbits was cultured.The synovial membrane was treated with 10 nM of 1,25(OH)2D3 for different lengths of time and the PRG4 gene expression was evaluated using quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR).PRG4 concentrations in the synovial membrane culture supernatant were measured using sandwich enzyme linked immunosorbent assay (ELISA).The optimal time point was determined for the subsequent experiments.After the synovial membrane was treated with different concentrations of 1,25(OH)2D3,the effect of 1,25(OH)2D3 on PRG4 gene expression and secretion was examined.Results Real-time PCR showed that PRG4 mRNA was up-regulated 6 h after 1,25(OH)2D3 treatment,and that PRG4 mRNA expression peaked at 24 h.ELISA results showed that PRG4 protein content in the synovial medium began to increase significantly 12 h after 1,25(OH)2D3 treatment,and reached the peak at 48 h.Real-time PCR analysis showed that 1,25(OH)2D3 promoted the expression of PRG4 gene in the joint synovial membrane within the range of 0.1-100 nM.The expression of PRG4 gene was increased gradually with 1,25(OH)2D3 concentrations.ELISA analysis showed that 1,25(OH)2D3 promoted the synovial secretion of PRG4 within the range of 0.1-100 nM.The synovial secretion of PRG4 gradually increased along with 1,25(OH)2D3 concentrations.Conclusions Our study has demonstrated that 1,25-(OH)2D3 can promote the PRG4 gene expression and secretion of synovial membrane.
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