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透明质酸对体外培养大骨节病软骨细胞增殖与凋亡的影响

期刊名称：中国地方病学杂志 2010;29(2):158-161
时间：0
分类：R681.3[医药卫生—骨科学;医药卫生—临床医学;医药卫生—外科学]
作者机构：[1]西安交通大学医学院第二附属医院骨科,710004, [2]西安交通大学医学院第二附属医院环境与疾病相关基因教育部重点实验室环境与地方病研究室,710004, [3]陕西省地方病防治研究所, [4]上海其胜生物制剂有限公司
相关基金：科技部国际合作重点项目（2006DFA33610）;国家自然科学基金（30630058）;陕西省国际合作重点项目（2005KW-13）
相关项目：大骨节病病因及发病机制研究

关键词：透明质酸, 大骨节病, 软骨细胞, 细胞增殖, 细胞凋亡, Hyaluronic acid , Kashin-Beck disease , Chondrocyte , Proliferation, Apoptosis

中文摘要：

目的通过观察透明质酸（HA）对体外培养的大骨节病（Kashin—Beck disease，KBD）软骨细胞增殖、凋亡的影响．为临床上HA治疗KBD提供实验依据。方法依据《大骨节病诊断标准》（GB16003—1995）收集KBD患者和遭遇意外事故的病人（对照组）关节软骨，分离、体外培养关节软骨细胞，选用第2代细胞进行实验。两组软骨细胞分别给予不同剂量的HA，按HA剂量分为0、100、500mg／L组，通过二苯甲唑溴盐（MTT）实验．测定第2．4、6天HA对KBD组、对照组软骨细胞增殖的影响。并通过流式细胞检测观察HA对软骨细胞凋亡的影响。结果对照组在第4天时，500mg／L组（0．140±0．049）促软骨细胞增殖作用大于0mg／L组（0．116±0．021）；KBD组在第6天时，500mg／L组（0．179±0．081）与0mg／L组（0．128±0．017）比较，显示了明显的促增殖作用（P〈0．05）。KBD组细胞凋亡率100、500mg／L组（10．458±1．143、7．877±1．346）均较0mg／L组（12．860±2．159）下降（P〈0．05）；对照组500mg／L组（4．045±1．204）较0mg／L组（7．128±1．244）细胞凋亡率下降（P〈0．05）。结论HA对KBD软骨细胞具有促进增殖和抑制软骨细胞凋亡的作用，其中500mg／L的HA改善KBD软骨细胞代谢的作用较100mg／L明显。

英文摘要：

Objective To understand the effect of hyaluronic acid（HA） on the proliferation and apoptosis of chondrocytes cultured in vitro with Kashin-Beck disease （KBD） to provide the experimental evidences for treating KBD diseases with HA. Methods The articular cartilage samples collected from KBD patients were selected according to Diagnosis for Kaschin-Beck Disease（GB 16003-1995）. And the normal cartilage samples were collected from victims of incidence （control）. Chondrocytes were separated and cultured in vitro. Then varying dosages of HA were administered to chondrocytes and individed into 0,100,500 mg/L group, according to HA doages. The effect of HA on the proliferation and apoptosis of chondrocytes cultured in vitro both KBD and the controls were investigated by methyl thiazolyl tetrazolium（MTT）, Annexin V/PI staining on 2^sd, 4^th, 6^th day. Results In the control group, 500 mg/L group （0.140 ± 0.049） promoted chondrocyte proliferation significantly than 0 mg/L group （0.116± 0.021） at the 4^th day（P 〈 0.05）, similar phenomenon was observed in KBD group in the 6^th day between 500 and 0 mg/L group（0.179 ± 0.081,0.128 ± 0.017, P 〈 0.05）. In the KBD group, compared with 0 mg/L （12.860 ± 2.159）, both 100 and 500 mg/L（10.458± 1.143,7.877± 1.346） inhibited chondrocyte apoptosis rate （P 〈 0.05 ）. In control, apoptosis rate of 500 mg/L group（4.045 ±1.204） descreased compared with 0 mg/L group （7.128± 1.244, P 〈 0.05）. Conclusion HA can promote the proliferation and inhibit the apoptosis of KBD chondrocytes cultured in vitro, and 500 mg/L HA play more effective role than that of 100 mg/L in promoting proliferation and inhibiting poptosis.

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