中文摘要：

目的观察透骨消痛胶囊含药血清对经TNF-α诱导的软骨细胞Wnt／β-catenin信号通路的影响，探讨其对关节软骨细胞的多靶点保护作用。方法将4周龄SD大鼠关节分离软骨细胞进行体外培养、传代，用免疫组化法鉴定第三代软骨细胞。以第3代软骨细胞为靶细胞，分为正常组、模型组和治疗组，模型组和治疗组用TNF-α诱导软骨细胞凋亡后，分别给予生理盐水血清和透骨消痛胶囊含药血清作用24h，Westernblot法观察各组软骨细胞中Wnt／β-catenin信号通路关键蛋白wnt4、β-catenin和GSK-3β的表达。结果（1）细胞鉴定结果为Ⅱ型胶原免疫细胞化学染色阳性，由此说明分离细胞为软骨细胞。（2）与正常组相比，模型组wnt4和β-catenin表达升高（P〈0．05），而GSK-3β的表达降低（P〈0．05）；与模型组相比，治疗组Wnt4和β-catenin表达降低（P〈0．05），而GSK-3β的表达升高（P〈0．05）。结论透骨消痛胶囊能多靶点调控Wnt／β-catenin信号通路，对软骨细胞具有保护作用，为其治疗骨关节炎提供依据。

英文摘要：

Objective To observe the effects of serum containing Tougu Xiaotong capsule （TGXTC） on Wnt/β-catenin signaling pathway in chondrocytes induced by TNF-α and discuss its multi-target protection effects on chondrocytes. Methods ArtiCular chondrocytes were isolated from Knee cartilages of four-week-old SD rats, cultured and subcultured in vitro. The third generation cells were identified by the methods of type Ⅱ collagen enzyme immunohistochemistry, and divided into three groups including normal group, model group and treatment group. After the chondrocytes of model group and treatment group were induced by TNF-α, they were intervened by serums containing physiological saline and TGXTC for 24 hours, respectively. Western blot was used to observe the expressions of key proteins Wnt 4, β-catenin and GSK-3β of Wnt/β-catenin signaling pathway in chondrocytes. Results The cells appeared positive after type Ⅱ collagen enzyme immunohistochemistry. This indicated they were chondroeytes. Compared with normal group, the expressions of Wnt 4 and β-catenin were significantly up-regulated in model group （P〈0.05）, but the expression of GSK-3β was significantly reduced in model group （P〈0.05）. Compared with model group, the expressions of Wnt 4 and β-catenin were significantly reduced in treatment group （P〈0.05）, but the expression of GSK-3β was significantly up-regulated in treatment group （P〈0.05）. Conclusions TGXTC has multi-target effects on regulating Wnt/β-catenin signaling pathway to protect chondrocytes, which provides evidence for treating osteoarthritis of TGXTC.