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目的:观察透骨消痛胶囊含药血清对硝普钠造模后软骨细胞TGF-β(转化生长因子-β)信号通路中ALK1,ALK5,ALK1/ALK5及MMP-13的表达影响,探讨其防治骨关节炎的作用机制。方法:2月龄清洁级雄性SD大鼠90只,随机分为空白组30只、壮骨关节丸组30只和透骨消痛胶囊组30只,制备含药血清。4周龄清洁级雄性SD大鼠膝关节软骨建立软骨细胞体外培养体系。1 mmol/L SNP干预第2代软骨细胞复制软骨细胞凋亡模型。分为1 mmol/L SNP组,1 mmol/L SNP+10%壮骨关节丸组,1 mmol/L SNP+5%、10%和15%透骨消痛胶囊组,分别培养3,6和9d,Real-time PCR和Western blot分别检测各组软骨细胞ALK1,ALK5及MMP-13基因及蛋白表达。结果:含药血清分别干预细胞3,6和9d后,ALK1,ALK1/ALK5及MMP-13基因及蛋白的表达,壮骨关节丸组和透骨消痛胶囊组均明显低于1 mmol/L SNP组(P<0.01,P<0.05);ALK5基因及蛋白的表达,壮骨关节丸组和透骨消痛胶囊组均明显高于1 mmol/L SNP组(P<0.01,P<0.05)。结论:透骨消痛胶囊通过调节TGF-β信号通路发挥抗软骨细胞凋亡作用。
OBJECTIVE: To observe the effect of the serum containing traditional Chinese medicine TouJiaoJiaoTong capsule on the expressions of ALK1, ALK5, ALK1 / ALK5 and MMP-13 in TGF-β (TGF-β) signaling pathway of rat model of sodium nitroprusside Prevention and treatment of osteoarthritis mechanism. Methods: Ninety clean male Sprague-Dawley rats of 2 months old were randomly divided into blank control group (n = 30), Zhuanggu Keli group (n = 30) and Tonggu Xiao Tong Capsule group (n = 30). Chondrocytes culture system was established in knee joint cartilage of 4-week-old clean male SD rats. Effect of 1 mmol / L SNP on Chondrocyte Apoptosis in 2nd Generation of Chondrocytes. The rats were divided into 1 mmol / L SNP group, 1 mmol / L SNP + 10% Zhuoguganjiewan group, 1 mmol / L SNP + 5%, 10% and 15% Real-time PCR and Western blot were used to detect the expression of ALK1, ALK5 and MMP-13 in chondrocytes respectively. RESULTS: The expressions of ALK1, ALK1 / ALK5 and MMP-13 mRNA and protein in the serum of 3 groups were significantly lower than those of 1 mmol / L SNP group (P <0.01, P <0.05). The expression of ALK5 gene and protein were significantly higher in Zhuanggu Keliwan and Touguixiao Capsules than those in 1 mmol / L SNP (P <0.01, P <0.05) . Conclusion: The traditional Chinese medicine TouJiaoJiaoTong capsule can prevent the apoptosis of chondrocytes by regulating the TGF-β signaling pathway.