丹参注射液对维甲酸诱导的软骨细胞凋亡的影响

目的:观察中药复方丹参注射液对全反式维甲酸在体外诱导的兔软骨细胞凋亡的调节作用.方法:取体外培养的兔软骨细胞,分别加入不同浓度维甲酸,维甲酸+丹参注射液,维甲酸+DEVD-FMK混合培养.并于培养后1、2、3、4天用DNA电泳,荧光显微镜和流式细胞仪观察软骨细胞凋亡变化.结果:在体外培养兔软骨细胞加维甲酸10-4M 24小时,流式细胞仪检测发现,凋亡率26.5±3.87%,而对照组为5±2.16% (P<0.01).荧光染色可见典型凋亡细胞形态改变,DNA电泳可见典型梯状条带,同时维甲酸诱导的软骨细胞凋亡具有剂量和时间依赖性,随着维甲酸浓度升高,时间延长,诱导凋亡作用明显增加.而维甲酸+丹参注射液或+DEVD-FMK混合培养的软骨细胞,其凋亡率明显下降.结论:维甲酸在体外能诱导兔软骨细胞凋亡,丹参注射液能降低维甲酸诱导的软骨细胞凋亡活性,提示其可能是治疗骨关节炎的机制之一.

Effects of Salvia Miltiorrhiza Injection on Retinoic Acid Induced Apoptosis in Chondrocyte

Objective:To observe the regulative effects of Salvia Miltiorrhiza injection on rabbit articular chondrocytes apoptosis induced by all trans retinoic acid ( RA ) with monolayer culture technique.Methods:Rabbit chondrocytes were cultured with various contrentrations of RA,RA and Salvia Miltiorrhiza injection,RA and DEVD-FMK for 1,2,3,4 days respectively.by Agarose gel electrophoresis of DNA ladder,flowcytometric analysis and flurorescent stain for cell were detected to determine the conditions of cellular apoptosis.Results:When chondrocytes were incubated with RA 10-4M for 24h,flowcytometric analysis showed that apoptotic cells amounted to 26.5±3.87%,while control was only 5±2.16% (P<0.01).Examination with fluorescence microscopy showed typical morphological change of apoptosis and electrophoresis showed the characteristic DNA ladder pattern.Induced apoptosis in chondrocytes by RA was dose and time dependent,and when the concentration of RA increased it expressed stronger activity in induced apoptosis.At the same time Salvia Miltiorrhiza injection and DEVD-FMK were found to down-regulate apoptosis in chondrocytes induced by RA.Conclusion:RA can induced apoptosis in chondrocytes in vitro,and Salvia Miltiorrhiza injection can decrease apoptosis induced by RA.This implies that decreasing apoptosis in chondrocytes may be the role of Salvia Miltiorrhiza injection in the treatment of osteoarthritis.