1,25(OH)2D3对关节滑膜蛋白聚糖-4基因表达及分泌的影响

王志强; 关超玲; 朱辉; 李贤慧

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武警医学 ›› 2021, Vol. 32 ›› Issue (1) : 42-45.

论著

1,25(OH)₂D₃对关节滑膜蛋白聚糖-4基因表达及分泌的影响

王志强¹, 关超玲², 朱辉³, 李贤慧²

作者信息 +

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

Author information +

文章历史 +

摘要

目的探讨维生素D体内活性形式1,25(OH)₂D₃对关节滑膜蛋白聚糖-4 (proteoglycan 4,PRG4)基因表达及分泌的影响。方法新西兰大白兔,分离膝关节滑膜体外培养。首先用10 nM浓度1,25(OH)₂D₃进行试验。Real time-PCR检测PRG4基因表达,ELISA 法检测培养液中PRG4分泌量的变化,确定1,25(OH)₂D₃作用后PRG4基因表达和PRG4分泌的最佳时间点。之后用不同浓度1,25(OH)₂D₃进行刺激,在最佳时间点测定1,25(OH)₂D₃对关节滑膜PRG4基因表达及分泌的影响。结果 Real time-PCR分析显示,1,25(OH)₂D₃作用6 h后PRG4 mRNA水平开始上调,24 h时PRG4 mRNA表达水平达峰值。ELISA分析结果显示,1,25(OH)₂D₃作用12 h时滑膜培养液中PRG4蛋白含量开始明显升高,48 h时滑膜培养液中PRG4蛋白含量达最高。Real time-PCR分析显示,在0.1~100 nM 浓度范围内1,25(OH)₂D₃均促进关节滑膜PRG4基因表达,随着1,25(OH)₂D₃浓度的提高,PRG4基因表达上调逐渐提高。ELISA分析显示,1,25(OH)₂D₃在0.1~100 nM浓度范围内均促进关节滑膜分泌PRG4,随着1,25(OH)₂D₃浓度的提高,滑膜分泌PRG4量逐渐增加。结论 1,25(OH)₂D₃对滑膜PRG4基因表达及分泌有明显的促进作用。

Abstract

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.

导出引用

王志强, 关超玲, 朱辉, 李贤慧. 1,25(OH)₂D₃对关节滑膜蛋白聚糖-4基因表达及分泌的影响[J]. 武警医学. 2021, 32(1): 42-45

WANG Zhiqiang, GUANG Chaoling, ZHU Hui, LI Xianhui. Influence of 1,25(OH)₂D₃ on proteoglycan 4 gene expression and secretion by rabbit synovial membrane[J]. Medical Journal of the Chinese People Armed Police Forces. 2021, 32(1): 42-45

中图分类号： R34 R68

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