少阳生骨方调节C518大鼠膝关节退变软骨细胞Ⅱ型胶原和基质金属蛋白酶13的表达

Shaoyang Shenggu formula regulates the expression of collagen type Ⅱ and matrix metalloproteinase 13 in degenerative cartilage cells of C518 rat's knee joint

背景:少阳生骨方有很好的治疗骨关节炎的作用,但其作用机制仍不清楚,可能与软骨退变的调节有关。目的:探讨少阳生骨方对C518细胞Ⅱ型胶原和基质金属蛋白酶13表达的影响。方法:选用C518大鼠膝关节退变软骨细胞,并用苏木精-伊红染色观察C518细胞形态;然后将C518细胞分为:少阳生骨方组、盐酸氨基葡萄糖组、生理盐水组,每组给予相应的干预后,再用RT-q PCR和Western blot检测Ⅱ型胶原和基质金属蛋白酶13基因和蛋白的表达水平。结果与结论:(1)RT-q PCR和Western blot检测结果显示,相较于生理盐水组,少阳生骨方组和盐酸氨基葡萄糖组Ⅱ型胶原的m RNA和蛋白表达量均上调(P<0.05),基质金属蛋白酶13的m RNA和蛋白表达量均下调(P<0.05);且相比盐酸氨基葡萄糖组,少阳生骨方组Ⅱ型胶原的m RNA和蛋白表达量显著上调(P<0.05),基质金属蛋白酶13 m RNA和蛋白表达量显著下调(P<0.05);(2)结果说明,少阳生骨方能调节C518大鼠膝关节退变软骨细胞中Ⅱ型胶原、基质金属蛋白酶13的m RNA和蛋白的表达水平,从而延缓退变的发展。

BACKGROUND： Shaoyang Shenggu formula is an effective prescription for osteoarthritis, which probably works via regulating cartilage degeneration, but the mechanism of action still needs an in-depth study.OBJECTIVE： To explore the effects of Shaoyang Shenggu formula on the expression of collagen type II and matrix metalloproteinase 13（MMP-13） in degenerative cartilage cells of C518 rat＇s knee joint. METHODS： C518 degenerative cartilage cells were selected, and then observed through hematoxylin-eosin staining. Afterwards, the cells were treated with Shaoyang Shenggu formula, glucosamine hydrochloride and normal saline, respectively. The expression levels of collagen type II and MMP-13 were detected by RT-q PCR and western blot assay. RESULTS AND CONCLUSION： Compared with normal saline, Shaoyang Shenggu formula and glucosamine hydrochloride significantly up-regulated the m RNA and protein expression levels of collagen type II, and significantly down-regulated the m RNA and protein expression levels of MMP-13. Moreover, the regulatory effects were more obvious in the group of Shaoyang Shenggu formula（P〈0.05）. To conclude, Shaoyang Shenggu formula delays the articular cartilage degeneration of the rat knee through regulating the expression of collagen type II and MMP-13 in C518 degenerative cartilage cells.