透骨消痛胶囊含药血清对软骨细胞外基质表达的影响**◆

背景：软骨细胞代谢异常引起细胞外基质降解和修复失衡是骨性关节炎的重要病理变化。透骨消痛胶囊具有补肾柔肝、活血祛风的功效,临床实验证实其对防治骨性关节炎有较好疗效。目的：观察透骨消痛胶囊含药血清对软骨细胞外基质表达的影响。方法：取SD大鼠膝关节软骨建立体外培养的软骨细胞,第3代软骨细胞同步化后进行干预。50只SD大鼠抽签法随机分为5组,用药量按照人体与动物药物等效剂量换算,空白组：生理盐水进行灌胃;对照组：壮骨关节丸水溶液灌胃;实验组：透骨消痛胶囊水溶液按0.145,0.290,0.580 mg/(g•d)进行灌胃。各组均连续给药3 d,采血前禁食12 h,末次给药1 h后腹主动脉采血,制备血清。结果与结论：Ⅱ型胶原免疫组织化学染色可见透骨消痛胶囊实验组Ⅱ型胶原表达强于空白组及对照组;透骨消痛胶囊实验组CollagenⅡ、蛋白聚糖、Aggrecan mRNA基因表达明显升高(P < 0.01),Cathepsin K mRNA基因表达明显降低(P < 0.01)。提示透骨消痛胶囊含药血清能上调软骨细胞CollagenⅡ、蛋白聚糖、Aggrecan mRNA基因表达,下调Cathepsin K mRNA基因表达,促进细胞外基质合成,调控细胞外基质和软骨细胞之间的动态平衡。关键词：透骨消痛胶囊;骨性关节炎;含药血清;软骨细胞;细胞外基质doi:10.3969/j.issn.1673-8225.2012.21.004

Effects of serum containing Tougu Xiaotong capsule on the expression of cartilage extracellular matrix

BACKGROUND: Osteoarthritis causes the abnormal metabolism of chondrocytes, which will further disorder the degradation and restoration balance of the extracellular matrix. Tougu Xiaotong capsule has been proved to have a good efficacy on the treatment of osteoarthritis.OBJECTIVE: To explore the effects of serum containing Tougu Xiaotong capsule on the expression of cartilage extracellular matrix. METHODS: Chondrocytes obtained from the knee joints of SD rats were cultured in vitro, and the third passage chondrocytes were intervened with serum containing Tougu Xiaotong capsule after synchronization. Fifty SD rats were divided into five groups randomly; the dose of the injection was conversed according to the drugs equivalent of human and animal. In the blank group, the rats were lavaged with normal saline; in the control group, the rats were lavaged with Zhuangguguanjie aqueous solution; in the experimental group, the rats were lavaged with 0.145, 0.290 and 0.580 mg/g per day Tougu Xiaotong capsule aqueous solution. The injection in each group lasted for 3 days, and fasting was preformed at 12 hours before blood collection. At 1 hour after the last administration, the abdominal aortic blood was collected and the serum was prepared.   RESULTS AND CONCLUSION: Immunohistochemical staining of collagen type Ⅱ showed that the expression of collagen Ⅱ in the experimental group was stronger than that in the blank and control groups. In the experimental group, the mRNA and protein expression of collagen Ⅱ, proteoglycan and Aggrecan was significantly increased (P < 0.01), while the mRNA and protein expression of Cathepsin K was significantly decreased (P < 0.01). Serum containing Tougu Xiaotong capsule can increase the mRNA and protein expression of collagen Ⅱ, proteoglycan, and Aggrecan in chondrocytes, decrease the mRNA and protein expression of Cathepsin K, promote the synthesis of extracellular matrix, and regulate the dynamic equilibrium between extracellular matrix and chondrocytes.