Yang Tao, Sun Wei, Duan Yuan-Hui, Sun Yun-Bo, Ren Yi-Ming, Hou Wei-Yu, Tian Meng-Qiang
Department of Joint and Sports Medicine, Tianjin Union Medical Center, Tianjin 300121, P.R. China.
The Postdoctoral Research Station, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, P.R. China.
Exp Ther Med. 2020 Aug;20(2):1775-1781. doi: 10.3892/etm.2020.8839. Epub 2020 Jun 4.
Low expression levels of 25-hydroxyvitamin D (vitamin D3) in the blood have been reported to be associated with the progression of osteoarthritis; however, the mechanisms by which this occurs remain unclear. The present study aimed to determine the effects of vitamin D3 on chondrocytes. MTT assays were used to determine whether vitamin D3 affects chondrocytes viability. Primary chondrocytes were treated with control culture medium, vitamin D3, tumor necrosis factor (TNF)-α, TNF-α + PNU-74654 [Wingless-related integration site (Wnt)/β-catenin signaling pathway inhibitor] or TNF-α + vitamin D3. Reverse transcription-quantitative PCR and western blotting were utilized to measure the gene and protein expression of collagen II, aggrecan, matrix metalloproteinase (MMP)-3 and MMP-13, A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, ADAMTS-5, Wnt-3a and nuclear β-catenin. The results demonstrated that TNF-α reduced the expression levels of aggrecan and collagen II, and increased the expression levels of MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5. Furthermore, vitamin D3 and PNU-74654 were observed to partially attenuate the effects induced by TNF-α. Moreover, similar findings were reported following co-treatment with vitamin D3 and TNF-α. Western blotting data revealed that TNF-α increased Wnt-3a and β-catenin protein levels in chondrocytes, while Vitamin D3 and PNU-74654 decreased the expression levels of Wnt-3a and nuclear β-catenin. In conclusion, the findings of the present study provided evidence to suggest that vitamin D3 may prevent articular cartilage degeneration and osteoarthritic disease progression by inhibiting the expression levels of MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5 through suppressing the Wnt/β-catenin signaling pathway. These results suggested that vitamin D3 may be of therapeutic value for the prevention and treatment of osteoarthritis.
据报道，血液中25-羟基维生素D（维生素D3）水平低与骨关节炎的进展有关；然而，其发生机制尚不清楚。本研究旨在确定维生素D3对软骨细胞的影响。采用MTT法测定维生素D3是否影响软骨细胞活力。原代软骨细胞分别用对照培养基、维生素D3、肿瘤坏死因子（TNF）-α、TNF-α + PNU-74654[无翅相关整合位点（Wnt）/β-连环蛋白信号通路抑制剂]或TNF-α + 维生素D3处理。利用逆转录定量PCR和蛋白质印迹法检测Ⅱ型胶原、聚集蛋白聚糖、基质金属蛋白酶（MMP）-3和MMP-13、含血小板反应蛋白基序的解聚素和金属蛋白酶（ADAMTS）-4、ADAMTS-5、Wnt-3a和核β-连环蛋白的基因和蛋白表达。结果表明，TNF-α降低了聚集蛋白聚糖和Ⅱ型胶原的表达水平，并增加了MMP-3、MMP-13、ADAMTS-4和ADAMTS-5的表达水平。此外，观察到维生素D3和PNU-74654可部分减轻TNF-α诱导的作用。此外，维生素D3与TNF-α联合处理后也有类似发现。蛋白质印迹数据显示，TNF-α增加了软骨细胞中Wnt-3a和β-连环蛋白的蛋白水平，而维生素D3和PNU-74654降低了Wnt-3a和核β-连环蛋白的表达水平。总之，本研究结果表明，维生素D3可能通过抑制Wnt/β-连环蛋白信号通路来抑制MMP-3、MMP-13、ADAMTS-4和ADAMTS-5的表达水平，从而预防关节软骨退变和骨关节炎疾病进展。这些结果表明，维生素D3可能对骨关节炎的预防和治疗具有治疗价值。
