威灵仙提取物干预膝骨关节炎软骨细胞的生长活力

Effect of radix clematidis extract on viability of knee osteoarthritis chondrocytes

背景:近年来研究显示关节软骨细胞过度凋亡是骨性关节炎开始和进展的关键因素,利用药物抑制软骨细胞凋亡来控制骨性关节炎的进展是现在的研究热点。而威灵仙在国内临床治疗骨关节炎时取得了较确切的疗效,但是其具体的作用机制是否通过抑制软骨细胞凋亡来实现尚未见相关报道。目的:观察中药威灵仙提取物对膝骨关节炎软骨细胞生长活力的影响。方法:取骨关节炎晚期行膝关节置换的关节软骨,剪碎后,采用酶消化法消化细胞,将处于对数生长期第3代细胞随机分组,实验组分别加入0.01,0.05,0.1,0.5,1.0g/L威灵仙提取物培养基,对照组仅加入普通培养基进行培养。Live/Dead染色法测定人软骨细胞活力指数,TUNEL染色法测定人软骨细胞凋亡指数。结果与结论:0.05,0.1g/L威灵仙提取物能明显提高软骨细胞活力,而0.5,1.0g/L威灵仙提取物反而降低了软骨细胞活力,其活力指数与对照组比较差异均有显著性意义(P<0.05);0.01,0.05,0.1g/L威灵仙提取物能有效抑制软骨细胞凋亡,1.0g/L威灵仙提取物反而促进了软骨细胞凋亡,其凋亡指数与对照组比较差异均有显著性意义(P<0.05)。结果可见威灵仙提取物在合适的质量浓度时能有效提高人软骨细胞活力,抑制人软骨细胞凋亡,尤以0.1g/L时效果最佳。

BACKGROUND： In recent years, studies have shown that excessive apoptosis of articular chondrocytes is the key factor for the start and progress of osteoarthritis. To study the progress in the use of drugs that inhibit apoptosis of chondrocytes to control osteoarthritis has become a hotspot. Clematis has obtained exact effect in the domestic clinical treatment of osteoarthritis, but the specific mechanism underlying inhibition of chondrocytes apoptosis has not been reported. OBJECTIVE： To observe the effect of radix clematidis extract on cell viability of the knee osteoarthritis chondrocytes.METHODS： Joint cartilage was shredded after harvested from the patients of osteoarthritis undergoing the knee replacements, and chondrocytes were isolated and cultured by the way of enzymatic digestion. The third-passage cells in the logarithmic growth phase were cultured in vitro and randomly divided into six groups after adherence. The experimental groups were cultured in Dulbecco’s modified Eagle’s medium with 0.01, 0.05, 0.1, 0.5, and 1.0 g/L radix clematidis extract, while the control group was given normal medium alone. Live/Dead assay method was adopted to observe the effect of radix clematidis extract with different concentrations on cell viability of the knee osteoarthritis chondrocytes, and TUNEL method was used to assay apoptosis index of the knee osteoarthritis chondrocytes.RESULTS AND CONCLUSION： 0.05 and 0.1 g/L radix clematidis extracts increased cell viability of chondrocytes, while 0.5 and 1.0 g/L radix clematidis extracts decreased the cell viability of chondrocytes. There was a significant difference in the cell viability between 0.05, 0.1, 0.5, 1.0 g/L radix clematidis extract groups and the control group （P 〈 0.05）. 0.01, 0.05, 0.1 g/L radix clematidis extracts effectively inhibited apoptosis of chondrocytes, while 1.0 g/L radix clematidis extract promote the apoptosis of chondrocytes. There was a significant difference in the apoptotic index between 0.01, 0.05, 0.1, 1.0 g/L radix clematidis extract groups and the control group （P 〈 0.05）. The appropriate concentration of radix clematidis extract could improve chondrocytes viability and restrain chondrocytes apoptosis, and the peak was at 0.1 g/L group. But excessive concentration of radix clematidis extract could reduce chondrocytes viability and promote chondrocytes apoptosis, representing the toxic effects on human articular chondrocytes.