Title1alpha, 25(OH)_2 D_3对大鼠关节软骨细胞蛋白聚糖和蛋白聚糖酶代谢的调节作用
Other TitlesRole of 1alpha, 25(OH)_2D_3in the metabolism of aggrecan and aggrecanase in chondrocytes of rat articular cartilage
Authors官剑
谭启钊
赵振达
刘忠军
宋纯理
冷慧杰
Affiliation北京大学第三医院骨科, 北京 100191, 中国
Keywords维生素D
蛋白聚糖
蛋白聚糖酶
软骨细胞
骨关节炎
1alpha
25(OH)_2D_3
aggrecan
aggrecanase
chondrocytes
osteoarthritis
Issue Date2018
Publisher中国实验动物学报
Citation中国实验动物学报. 2018, 26(3), 287-295.
Abstract目的在细胞水平,探讨1alpha, 25(OH)_2D_3对受到多种促炎症因子干预的大鼠关节软骨细胞蛋白聚糖合成和分解的调节作用。方法分别使用促炎症因子IL-1alpha、IL-1 beta及TNF-alpha对大鼠关节软骨细胞进行炎症诱导,使用不同剂量的1alpha, 25(OH)_2D_3对正常及炎症状态下的软骨细胞进行干预,使用CCK8、流式细胞术分析、real time-PCR及western blot等方法分别检测软骨细胞的增殖活性和凋亡水平,以及软骨细胞中蛋白聚糖和蛋白聚糖酶-1 /2即ADAMTS-4及ADAMTS-5的mRNA和蛋白表达水平变化,从而评估1alpha, 25(OH)_2D_3对大鼠关节软骨细胞蛋白聚糖和蛋白聚糖酶代谢的调节作用。结果IL-1alpha、IL-1beta及TNF-alpha显著降低 了软骨细胞的增殖活性并增加了软骨细胞的凋亡水平,以及降低细胞中蛋白聚糖的mRNA和蛋白表达水平,增加ADAMTS-4及ADAMTS-5的mRNA 和蛋白表达水平。1alpha, 25(OH)_2D_3的干预可以增加炎症状态下软骨细胞的增殖活性、降低软骨细胞的凋亡水平,以及增加炎症状态下软骨细胞内蛋白聚糖的mRNA和蛋白表 达水平,降低ADAMTS-4及ADAMTS-5的mRNA和蛋白表达水平,并呈一定剂量依赖性。但是,1 alpha, 25(OH) _2D_3的干预并不影响正常软骨细胞的增殖、凋亡。结论维生素D能促进炎症状态下的软骨细胞蛋白聚糖的合成代谢并抑制蛋白聚糖酶活性,从而为关节软骨提 供更全面的保护。
Objective To investigate the role of vitamin D in the synthesis and degradation of aggrecan in rat articular chondrocytes at cellular level. Methods Rat articular chondrocytes were stimulated by IL-1alpha,IL-1beta and TNF- alpha,respectively. Normal and inflammatory chondrocytes were treated with different doses of vitamin D,respectively. CCK8,Flow cytometry,real time-PCR and western blot analysis were used to examine the proliferation activity and apoptosis level of chondrocytes,and the expression of aggrecan,ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels. Results IL-1alpha, IL-1beta and TNF-alpha significantly decreased the proliferation activity and increased the apoptosis level of the chondrocytes. Furthermore,IL-1alpha,IL-1beta and TNF-alpha significantly decreased the expression of aggrecan,and increased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes. 1alpha,25 (OH) _2D_3 supplementation significantly increased the proliferation activity and decreased the apoptosis level of chondrocytes stimulated by IL-1alpha, IL-1beta and TNF-alpha in a dose-dependent manner, but not affected the normal chondrocytes. Meanwhile,1alpha,25 (OH) _2D_3 also significantly increased the expression of aggrecan,and decreased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes under inflammatory conditions. Conclusions Vitamin D may promote the anabolism of aggrecan and inhibit aggrecanase activity in chondrocytes under inflammatory conditions,which may impact overall protection for articular cartilage.
URIhttp://hdl.handle.net/20.500.11897/514811
ISSN1005-4847
Indexed中国科学引文数据库(CSCD)
Appears in Collections:第三医院

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