摘要
目的:利用体外软骨细胞培养体系,观察中药补肾活血方含药血清对软骨细胞增殖及蛋白质合成的影响,以探讨补肾活血中药防治骨性关节炎的作用。方法:实验于2003-05/2004-08在广州中医药大学实验动物中心完成。①4周龄新西兰兔,用于分离培养软骨细胞。②雄性SD大鼠10只,体质量250～300g,随机分为正常组和中药补肾活血方组,每组5只,用于制备含药血清。③将传一代的软骨细胞接种于96孔培养板和24孔培养板中,培养24h后细胞贴壁进行分组试验,培养孔随机分为6组,每组4孔:空白对照组(加体积分数为0.05血清DMEM)5%中药含药血清组,10%中药含药血清组,20%中药含药血清组,5%丹参注射液组,5%葛根素注射液组;用噻唑蓝法检测细胞活性和增殖,考马斯亮蓝法检测细胞总蛋白。结果:①5%,10%,20%中药含药血清组软骨细胞增殖与空白对照组相比有显著差异眼(1.10±0.04),(1.14±0.04),(1.14±0.06),(0.66±0.07),P<0.05演。②5%,10%,20%中药含药血清组细胞总蛋白合成与空白对照组相比有显著差异眼(137.60±5.98),(135.20±9.37),(130.80±2.50),(99.15±5.08),P<0.05演。③5%丹参及葛根素注射液组对软骨细胞增殖和蛋白合成无明显影响。结论:补肾活血中药含药血清可促进软骨细胞的增殖和蛋白合成,这可能是补肾活血中药防治骨性关节炎的重要作用机制之一。
AIM： To investigate the effect of Chinese herbs of bushen huoxue prescription on proliferation and the protein synthesis of chondrocytes cultured in vitro, so as to explore its preventive effect on osteoarthritis.
METHODS： The experiment was conducted in the Animal Laboratory of Guangzhou University of Traditional Chinese Medicine between May 2003 and August 2004. ① New Zealand rabbits with four weeks old were selected for isolation and culture of chondrocytes. ②Ten male SD rats with body mass of 300 300 g were selected and randomly divided into two groups： Normal group and busen huoxue prescription group with 5 rats in each group for preparation of serum containing Chinese herbs. ③The first subculture chondrocytes were plated in a 96 well plate and a 24 well plate, respectively. After 24 hours of incubation, when cell attachment was determined, cultured plates were divided into 6 groups with 4 well in each group： control group （added with 0.05 volume fraction DMEM）, serum groups containing 5%, 10% and 20% Chinese herb of bushen huoxue prescription, 5% danshen injection group and 5% gegen injection group： the cytoactive and proliferation were detected by methyl thiazolyl tetrazolium colorimetric assay. and cell total protein was assessed by coomassie brilliant blue G-250 colorimetric assay.
RESULTS： ①The proliferation of chondrocytes in the serum groups containing 5%, 10%, 20% of bushen huoxue prescription differed significantly compared with the control group [（1.10±0.04）, （1.14±0.04）, （1.14±0.06）, （0.66±0.07）, P 〈 0.01]. ②There were significant differences in cell total protein synthesis among the serum groups containing 5%, 10%, 20% herbs and the control group [（137.60±5.98）, （135.20±9.37）, （130.80±2.50）, （99.15±5.08）, P 〈 0.05]. ③ 5% danshen injection and 5% gegen injection had no obvious effect on the proliferation and protein synthesis of chondrocytes.
CONCLUSION： The serum containing Chinese herb of bushen huoxue prescription can promote the proliferation and protein synthesis of chondrocytes, which may be the important mechanism of bushen huoxue prescription in preventing and treating osteoarthritis.
出处
《中国临床康复》
CSCD
北大核心
2006年第19期50-52,共3页
Chinese Journal of Clinical Rehabilitation
基金
广东省自然科学基金资助(020791)
国家中医药管理局科学技术研究专项基金资助(02-03JP33)~~
