灵仙通络方对C518大鼠膝关节退变软骨细胞增殖、Ⅱ型胶原及基质金属蛋白酶-13表达的影响

摘要

目的：探讨灵仙通络方对C518大鼠膝关节退变软骨细胞增殖、Ⅱ型胶原（COL-Ⅱ）及基质金属蛋白酶-13（MMP-13）的影响。方法选取C518大鼠膝关节软骨细胞株，随机分为中药组（灵仙通络方）、西药组（氨糖美辛）和对照组（氯化钠溶液），进行培养、诱导退变等处理，于给药后第1、2、3 d，分别检测中药组、西药组和对照组不同浓度下对软骨细胞增殖、COL-Ⅱ及MMP-13表达的影响。结果同一时间点的中药组、西药组不同浓度之间噻唑蓝（MTT）值差异有统计学意义（P＜0.01）。对照组不同浓度氯化钠溶液的MTT值差异不具有统计学意义（P＞0.05）。在COL-Ⅱ免疫组化染色中，中药组与西药组和对照组相比差异有统计学意义（P＜0.01），且中药组优于西药组（染色指数χ2=16.26，df=2，P＜0.01）。在 MMP-13免疫组化染色中，对照组较中药组、西药组差异有统计学意义（P＜0.01），且中药组优于西药组（染色指数χ2=54.31，df=2，P＜0.01）。结论 C518大鼠膝关节退变软骨细胞经灵仙通络方干预后能够促进其增殖，调节COL-Ⅱ、MMP-13的表达，延缓膝关节骨性关节炎疾病的发展进程。%〔Abstract〕 Objective To investigate the effect of Lingxian Tongluo prescription on the expression of cartilage cell, COL-II and MMP-13 in C518 rats with knee joint degeneration. Methods Selected C518 rat knee cartilage cell line as the object, which were randomly divided into the Chinese medicine group (Lingxian Tongluo decoction) and Western medicine group (indometacin and glucosamine) and control group (sodium chloride solution), and then the above groups were treated with culture and induced degeneration. On the 1st d, 2nd d, 3rd d after administration, and then the expression of cartilage cell, COL-II and matrixmetalloproteinase-13 (MMP-13) in rats with knee joint degeneration was detected. Results At the same time point, the MTT values of the Traditional Chinese medicine group and Western medicine group had significant difference (P<0.01). The MTT values of the control group were not statistically significant (P>0.05). Compared with the Western medicine group and the control group, the COL-II immunohistochemical staining of Traditional Chinese medicine group were statistically significant (P<0.01), and the Traditional Chinese medicine group was better than the Western medicine group (dyeing index χ2=16.26, df=2, P<0.01). The MMP-13 immunohistochemistry staining in the control group was significant comparing with the Traditional Chinese medicine group and the Western medicine group, with statistical significance (P<0.05), and the Western medicine group compared with the Traditional Chinese medicine group was significantly different (P<0.05), and the traditional Chinese medicine group was better than that of the Western medicine group (dyeing index χ2=54.31, df=2, P<0.05). Conclusion The knee degenerative cartilage cells of C518 rats intervened by Lingxian Tongluo prescription can promote its proliferation, regulate the expression of COL-II and MMP-13, and delay the development of knee osteoarthritis disease.