CAJ | 학술논문

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0.01,0.05,0.1,0.5,1.0 g/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:0.05,0.1 g/L威灵仙提取物能明显提高软骨细胞活力,而0.5,1.0 g/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);0.01,0.05,0.1 g/L威灵仙提取物能有效抑制软骨细胞凋亡,1.0 g/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0.1 g/L时效果最佳.
배경:근년래연구현시관절연골세포과도조망시골성관절염개시화진전적관건인소,이용약물억제연골세포조망래공제골성관절염적진전시현재적연구열점.이위령선재국내림상치료골관절염시취득료교학절적료효,단시기구체적작용궤제시부통과억제연골세포조망래실현상미견상관보도.목적:관찰중약위령선제취물대슬골관절염연골세포생장활력적영향.방법:취골관절염만기행슬관절치환적관절연골,전쇄후,채용매소화법소화세포,장처우대수생장기제3대세포수궤분조,실험조분별가입0.01,0.05,0.1,0.5,1.0 g/L위령선제취물배양기,대조조부가입보통배양기진행배양.Live/Dead염색법측정인연골세포활력지수,TUNEL염색법측정인연골세포조망지수.결과여결론:0.05,0.1 g/L위령선제취물능명현제고연골세포활력,이0.5,1.0 g/L위령선제취물반이강저료연골세포활력,기활력지수여대조조비교차이균유현저성의의(P <0.05);0.01,0.05,0.1 g/L위령선제취물능유효억제연골세포조망,1.0 g/L위령선제취물반이촉진료연골세포조망,기조망지수여대조조비교차이균유현저성의의(P <0.05).결과가견위령선제취물재합괄적질량농도시능유효제고인연골세포활력,억제인연골세포조망,우이0.1 g/L시효과최가.
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 cel 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 cel s 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 cel 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 cel viability of chondrocytes, while 0.5 and 1.0 g/L radix clematidis extracts decreased the cel viability of chondrocytes. There was a significant difference in the cel 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.