独活寄生汤含药血清通过调控SOX6对体外软骨细胞表型的影响

Effect of Duhuo Jisheng Decoction on the Phenotype of Chondrocytes in Vitro by Regulating SOX6

关键词:  独活寄生汤  软骨细胞  转录因子SOX6

Key words:  Duhuo Jisheng decoction  chondrocytes  SOX6

摘要:

目的 研究大鼠独活寄生汤含药血清对体外软骨细胞表型稳定性，SOX6及培养基中细胞因子IL-1β、IL-6、TNF-α表达的影响。方法 取大鼠股骨头软骨帽，提取软骨细胞，并在体外单层传代培养。培养体系为含高、中、低浓度的独活寄生汤含药血清及生理盐水血清，通过免疫组织化学分析软骨细胞表型Ⅱ型胶原的变化及SOX6表达分布，通过Westernblot及ELISA检测SOX6、Collagen Ⅱ、Aggrecan及IL-1β、IL-6、TNF-α表达的变化。结果 不同浓度的独活寄生汤含药血清均可对体外培养的软骨细胞产生增殖促进作用，并能增加Ⅱ型胶原稳定其软骨表型；其中以中浓度最为显著（P<0.01）；与生理盐水血清组相比，独活寄生汤含药血清可以增加SOX6在细胞核的分布，而且能抑制培养基中细胞因子IL-1β、IL-6、TNF-α的表达水平。结论 独活寄生汤含药血清可以上调体外软骨细胞SOX6及抑制炎症因子的表达以稳定软骨细胞的表型。

Abstract:

OBJECTIVE To study the effects of serum containing Duhuo Jisheng decoction on the phenotypic stability of chondrocytes in vitro, and on the expression of SOX6, cytokines IL-1β, IL-6 and TNF-α. METHODS Cartilage caps were extracted from the femoral head of rats, and chondrocytes were isolated and cultured in vitro. The culture system were serum containing Duhuo Jisheng decoction (different doses of high, medium and low concentrations) and serum containing normal saline as control. The changes of collagen type Ⅱ and SOX6 expression distribution in chondrocytes were analyzed by immunohistochemistry. The changes of SOX6, Collagen Ⅱ, Aggrecan, IL-1 beta, IL-6, TNF-α were detected by Western blot and ELISA. RESULTS The serum containing Duhuo Jisheng decoction of different doses could promote the proliferation of chondrocytes in vitro, and increase the expression of collagen Ⅱ to stabilize its cartilage phenotype. The medium does showed most significant (P<0.01). Compared with the normal control group, the serum containing Duhuo Jisheng decoction could increase the fraction of SOX6 in the nucleus. It also inhibited the expression levels of cytokines IL-1β, IL-6 and TNF-α in the medium. CONCLUSION The serum containing Duhuo Jisheng decoction can upregulate the expression of SOX6 and inhibit the expression of inflammatory factors in vitro, so as to stabilize the phenotype of chondrocytes.