柔肝复方及单味药提取物对体外培养人骨关节炎软骨细胞增殖及关节软骨低聚基质蛋白分泌的研究

In vitro study the effect of both extract and compound from Liver-softening herbs on chondrocyte proliferation and COMP expression of human osteoarthritis

目的：探讨柔肝单味药提取物金利胶囊及复方养血软坚胶囊对体外培养人骨关节炎（OA）软骨细胞增殖能力及关节软骨低聚基质蛋白（COMP）分泌的影响。方法：分阶段酶消化法体外培养人骨关节炎软骨细胞,以1×10^5/cm^2密度接种3代内细胞,研究设正常对照组及金利胶囊与养血软坚胶囊2个药物干预组。药物直接添加体外培养体系,添加终浓度为10mg/ml。以MTS/PMS法观察不同药物对体外培养的软骨细胞在接种1、3、5d时增殖情况的影响,采用ELISA法检测药物对体外培养的人软骨细胞COMP分泌的影响。结果：与空白组相比,金利胶囊及养血软坚胶囊未表现出明显的促进人骨关节炎软骨细胞增殖能力（P〉0.05）,但在各用药组内,细胞增殖能力在1~3d时间段较3~5d时间段明显增强（P〉0.05）。各药物组有上调人OA软骨细胞分泌COMP的作用,但组间未见有明显差异（P〉0.05）。结论：用药干预后,柔肝单味药提取物金利胶囊及复方养血软坚胶囊可以促进人骨关节炎软骨细胞分泌COMP,细胞增殖能力在1~3d时间段较3~5d时间段明显增强,但2组药物之间未见有明显差异。

Objective： To investigate the effect of both extract（RGL） and compound（BNHS） from Liver-softening herbs on chondrocyte proliferation and cartilage oligomeric matrix protein （COMP） expression of human osteoarthritis chondrocyte cultured in vitro. Methods： Chondrocytes isolated by enzymatic digestion from human knee osteoarthritis cartilage were cultured within 3 generations with the density of 1 × 10^5/cm^2. There were one control group and two study groups treated with RGL and BNHS respectively in this study. After directly adding drugs 10 mg/ml to cuhures,chondrocyte proliferation was observed at 1 st,3 rd and 5th days by using the 3- （4,5-dimethylthiazol-2yl） -5- （3-carboxymethoxy-phenyl） -2- （4-sulfophenyl） -2H-tetrazoli- um inner salt （MTS） assay kit. COMP level from culture supematant was tested by enzyme-linked immunoabsorbent assays （ELISA） Results：Although both RGL and BNHS had no significant effects on chondrocyte proliferation as compared to the control group（P 〉0. 05） ,cell number in 1 to 3 days interval increased much mere rapidly than in 3 to 5 days interval（ P 〈 0. 05 ）in each group. Compared with the control group,both extract and compound group had high COMP level, but no significant difference was demonstrated between the two groups. Conclusion. The finding suggests that both extract and compound from Liver-softening herbs could promote COMP expression of chondrocyte in vitro, but significant effect of stimulating cell proliferation is not yet be validated.