透骨消痛胶囊含药血清对退变关节软骨细胞表达TNF-α、IL-1β的影响

Impacts of Medicated Serum of Tougu Xiaotong Capsules on the Expression of TNF-α and IL-1β of Chondrocytes in Degenerated Joints

目的观察透骨消痛胶囊含药血清对体外培养的大鼠退变软骨细胞表达肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)的影响,探讨透骨消痛胶囊防治骨性关节炎的作用机制。方法 4周龄SD大鼠膝关节软骨建立体外培养的软骨细胞,采用Ⅱ型胶原原位杂交法对第3代软骨细胞进行鉴定。用10 ng.mL-1IL-1β干预第3代软骨细胞复制退变软骨细胞模型。模型复制成功后,随机分为模型组和透骨消痛胶囊含药血清组,分别在培养24、48、72 h后用TaqMan real-time PCR法检测各组mRNA表达。结果透骨消痛胶囊含药血清组TNF-αmRNA、IL-1βmRNA相对表达量随着干预时间的延长逐渐下降,差异均有统计学意义(P<0.05)。结论透骨消痛胶囊治疗骨性关节炎的部分作用机制是有效抑制退变软骨细胞TNF-α、IL-1β的表达。

Objective To observe the impacts of the medicated serum of tougu xiaotong capsules on the expressions of tumor necrosis factor-α（TNF-α）and interleukin-1β（IL-1β）of chondrocytes of the degenerated joints cultured in vitro for the rats and explore the mechanism of tougu xiaotong capsules on the prevention of treatment of osteoarthritis.Methods The chondrocytes were cultured in vitro in the cartilage of the knee joints for the 4-week old SD rats.II-type collagen in-situ hybridization（ISH）method was adopted to identify the chondrocytes of the 3rd generation.IL-1β,10ng/mL was used in the intervention of the chondrocytes of the 3rd generation so as to duplicate the model of the degenerated chondrocytes.After the model was duplicated successfully,in randomization,a model group and a medicated serum of tougu xiaotong capsule group（medicated serum group）were divided.Separately,in 24 h,48 h and 72 h of culture,TaqMan real-time PCR assay was used to detect mRNA expression in each group.Results In the medicated serum group,the relative expressions of TNF-α mRNA and IL-1β mRNA were reduced gradually with the time prolonging of the intervention.The difference was significant statistically（P0.05）.Conclusion Tougu xiaotong capsules inhibits effectively the expressions of TNF-α and IL-1β of the degenerated chondrocytes,which is the partial mechanism of it on the treatment of osteoarthritis.