透骨消痛颗粒对关节软骨细胞wnt/β-连环蛋白信号通路的调控

背景：由巴戟天、杭白芍、肿节风和川芎等组成的透骨消痛颗粒能有效延缓关节宏观形态、软骨基质及软骨细胞退变。目的：观察透骨消痛颗粒对软骨细胞wnt/β-连环蛋白信号通路中Wnt4、糖原合成酶3β及β-连环蛋白的影响。方法：将SD大鼠关节软骨细胞分离培养成功后，人白细胞介素 1β诱导软骨细胞退变，取第2代退变软骨细胞，随机分为对照组和透骨消痛颗粒组，后者加入透骨消痛颗粒醇提物，分别培养4，8 d后，采用RT-PCR检测2组Wnt4、糖原合成酶3β、β-连环蛋白mRNA表达，采用蛋白印迹法检测Wnt4、糖原合成酶3β及β-连环蛋白表达。结果与结论：采用人白细胞介素1β可诱导软骨细胞退变，软骨细胞退变早期均有Wnt4、糖原合成酶3β及β-连环蛋白表达，但随着时间推移Wnt4、β-连环蛋白表达下降，而糖原合成酶3β表达增高，采用透骨消痛颗粒醇提物干预后均可逆转上述现象。说明透骨消痛颗粒醇提物能诱导软骨细胞转录合成β-连环蛋白和Wnt4蛋白，并抑制糖原合成酶3β表达。

Regulatory effects of Tougu Xiaotong granule on wnt/β-catenin signal pathway of articular chondrocytes

BACKGROUND: Tougu Xiaotong granule composed of mornda root, radix paeoniae alba from Hangzhou of China, herba sarcandrae, and szechwan lovage rhizome can effectively delay alteration of macroscopic form of articular cartilage and degeneration of cartilage matrix and chondrocytes. But the underlying mechanism remains poorly understood. OBJECTIVE:To observe the effects of Tougu Xiaotong granule on Wnt4, glycogen synthase kinase 3β (GSK-3β) and β-catenin levels in wnt/β-catenin signal pathway of chondrocytes.  METHODS:SD rat articular chondrocytes were isolated and induced with human interleukin-1β for degeneration. Passage 2 degenerated chondrocytes were randomly divided into a control group and a Tougu Xiaotong granule group. Alcohol extract product of Tougu Xiaotong granule was added to the Tougu Xiaotong granule group. After 4 and 8 days of culture, Wnt4, GSK-3β and β-catenin levels were detected using RT-PCR and western blot methods. RESULTS AND CONCLUSION:Human interleukin-1β could induce the degeneration of chondrocytes. Wnt4, GSK-3β, β-catenin protein expression was observed in the chondrocytes during the early stage of degeneration. But with time going, Wnt4 and β-catenin protein expression was decreased while GSK-3β expression was increased. After intervention with Tougu Xiaotong granule, these phenomena could be reversed. These findings suggest that Tougu Xiaotong granule can induce chondrocytes to synthesize Wnt4 and β-catenin and inhibit GSK-3β expression.