骨碎补总黄酮对IL-1β诱导体外软骨细胞损伤的保护作用

Protective Effect of Total Flavones of Drynariae Rhizoma on IL-1β Induced Chondrocyte Injury in Vitro

关键词:  骨碎补总黄酮|软骨细胞|炎症|凋亡

Key words:  total flavones of Drynariae Rhizoma|chondrocyte|inflammation|apoptosis

摘要:

目的 探究骨碎补总黄酮（total flavones of Drynariae Rhizoma，TFDR）对IL-1β诱导的软骨细胞损伤的保护作用及相关机制。方法 分离并培养SD大鼠软骨细胞，采用CCK8法检测不同浓度的TFDR对软骨细胞增殖的影响以及TFDR对IL-1β诱导的软骨细胞增殖的影响；采用ELISA试剂盒检测TFDR对IL-1β诱导的软骨细胞中炎症因子的影响；采用Hoechst 33342染色法检测TFDR对IL-1β诱导的软骨细胞凋亡的影响；qRT-PCR和Western blotting检测凋亡相关因子和骨关节炎相关因子的表达情况。结果 不同浓度的TFDR能够在不同程度上促进软骨细胞的增殖；与对照组相比，IL-1β诱导的软骨细胞活性显著降低，炎症因子表达显著升高，细胞凋亡显著增加；IL-1β诱导的软骨细胞经TFDR干预后，细胞活性增加，炎症因子表达下降，细胞凋亡受到抑制。IL-1β诱导的软骨细胞中iNOS、COX-2、MMP-13、ADAMTS-5、Bax、Caspase-3 mRNA表达均显著增强，collagen-II、aggrecan、Bcl-2表达均显著降低；经TFDR处理后，iNOS、COX-2、MMP-13、ADAMTS-5、Bax、Caspase-3 mRNA表达降低，collagen-II、aggrecan、Bcl-2表达升高。结论 TFDR对IL-1β诱导的软骨细胞损伤具有一定的保护作用，其机制和调控炎症反应、抑制细胞凋亡相关。

Abstract:

OBJECTIVE To investigate the protective effect of total flavones of Drynariae Rhizoma(TFDR) on IL-1β induced chondrocyte injury and its related mechanism. METHODS The chondrocytes of SD rats were isolated and cultured. The effects of TFDR at different concentrations on chondrocytes proliferation and IL-1β induced chondrocytes proliferation were determined by CCK8 assay. The effects of TFDR on inflammatory cytokines in IL-1β induced chondrocytes were examined using ELISA kit; Hoechst 33342 staining was used to detect the effect of TFDR on IL-1β induced chondrocyte apoptosis; qRT-PCR and Western blotting were used to detect the expression of factors related to apoptosis and osteoarthritis. RESULTS Different concentrations of TFDR could promote the proliferation of chondrocytes in different degrees. Compared with control group, IL-1β induced chondrocytes were significantly reduced in activity, inflammatory factor expression was significantly increased, and apoptosis was significantly increased. After TFDR intervention, the proliferation activity were significantly increased, inflammatory factor expression was decreased, and apoptosis was inhibited. iNOS, COX-2, MMP-13, ADAMTS-5, Bax and Caspase-3 mRNA expression were significantly increased in IL-1β induced chondrocytes, while collagen II, aggrecan and Bcl-2 expression were significantly decreased. After TFDR treatment, expressions of iNOS, COX-2, MMP-13, ADAMTS-5, Bax and Caspase-3 mRNA were decreased, and collagen II and aggrecan and Bcl-2 expression were increased. CONCLUSION TFDR has a protective effect on IL-1β induced chondrocyte injury, and its mechanism is related to the regulation of inflammatory response and apoptosis.