硒对体外培养大骨节病软骨细胞生长及凋亡的影响  

Effect of selenium on proliferation and apoptosis of Kaschin-Beck disease chondrocyte cultured in vitro

作　　者：段琛[1] 郭雄[1] 张晓东[2] 高宗强[3] 张银刚[2] 于曰祥[4] 

机构地区：[1]西安交通大学医学院公共卫生系地方病研究所,710061 [2]西安交通大学医学辽第一附属医院外科,710061 [3]西安交通大学医学辽第二附属医院骨科,710061 [4]陕西省地方病防治研究所外科

出　　处：《中国地方病学杂志》2010年第5期480-484,共5页Chinese Jouranl of Endemiology

基　　金：科技部“十一五”支撑计划（2007BA125B004）;国家自然科学基金重点项目（306030058）

摘　　要：目的 观察硒对体外培养大骨节病（KBD）患者和正常人关节软骨细胞增殖和凋亡的影响,探索补硒防治KBD的作用,并为硒对正常软骨细胞生长的影响提供依据.方法 依据〈大骨节病临床诊断标准〉（GB 16003-1995）,选择Ⅱ度和Ⅲ度KBD患者5例和非病区正常人意外事故者5例的关节软骨进行体外分离、培养.KBD组和对照组分别给予不同剂量的硒（0、0.0125、0.0250、0.0500、0.1000、0.2500、0.5000、1.0000 mg/L）进行干预,采用四氮唑蓝（MTT）法、流式细胞仪和免疫组化法观察细胞生长和凋亡情况.结果 对照组第6天时各剂量组的细胞增殖率（0.086±0.025、0.077±0.012、0.073±0.027、0.071±0.017、0.058±0.028、0.052±0.028和0.046±0.037）比0 mg/L组（0.138±0.026）明显降低（P均＜0.05）;0.1000～1.0000 mg/L剂量组的平均细胞增殖率为负值（-0.001±0.001、-0.003±0.000、-0.003±0.001和-0.004±0.001）,显著低于0 mg/L组（0.025±0.003,P均＜0.05）;KBD组,与0 mg/L组（0.115±0.011）比较,0.2500 mg/L剂量组促进细胞增殖（0.128±0.037,P＜0.05）,1.0000 mg/L剂量组细胞生长受到抑制（0.071±0.019,P＜0.05）.对照组0.0500～1.0000mg/L剂量组的细胞凋亡率[（18.88±0.02）%、（17.58±0.01）%、（17.09±0.04）%、（56.00±0.02）%、（57.85±0.03）%]比0 mg/L组[（13.51±0.01）%]增高（P均＜0.05）;KBD组,与0 mg/L组[（25.84±0.02）%]比较,0.0250～0.2500 mg/L剂量组的细胞凋亡率[（13.69±0.02）%、（15.96±0.03）%、（16.68±0.03）%、（16.67±0.02）%]降低,0.5000、1.0000 mg/L剂量组的细胞凋亡率[（59.58±0.03）%、（73.48±0.04）%]明显增高（P均＜0.05）.KBD组0.0500～0.2500 mg/L剂量组的Fas表达[（41.2±1.5）%、（40.3±2.0）%、（50.2±2.5）%]低于同剂量硒干预的对照组[（52.4±1.0）%、（67.2±4.0）%、（75.1±5.0）%,P均＜0.05],0.0500、0.1000 mg/L剂量组的Caspase-3表达[（40.8±1.1）%、（45.1±2.1）%]低于同剂量硒干预的�Objective To investigate the effect of selenium on proliferation and apoptosis of chondrocytes of articular cartilage cultured in vitro in Kaschin-Beck disease（KBD） patients and normal person, to explore the role of selenium in control of KBD, and to provide evidence for selenium＇s effect on the growth of normal cartilage cells. Methods The articular cartilage samples of grade Ⅱ and Ⅲ KBD patients were selected according to the national ＂Clinical Diagnosis of KBD＂ （GB 16003-1995）. Chondrocytes of 5 KBD and 5 non-endemic normal accidentswere separated and cultured in vitro. KBD group and control group were given different doses of selenium （0,0.0125,0.0250,0.0500,0.1000,0.2500,0.5000,1.0000 mg/L, respectively）. Methyl thiazolyl tetrazolium （MTT）,flow cytometric analysis, and immunocytochemical staining were used to observe the effect of selenium on cell growth and apoptosis in KBD and normal persons. Results MTT results showed that the cell proliferation rate in each dosage group of the control group at the 6th day（0.086 ± 0.025,0.077 ± 0.012,0.073 ± 0.027,0.071 ± 0.017,0.058 ± 0.028,0.052 ± 0.028 and 0.046 ± 0.037） was significantly lower than that of 0 mg/L group（0.138 ± 0.026,all P 〈 0.05）;the average cell proliferation rate was negative（ - 0.001 ± 0.001, - 0.003 ± 0.000, - 0.003 ± 0.001and - 0.004 ± 0.001 ） in 0.1000 - 1.0000 mg/L dose group, which was significantly lower than that of the 0 mg/L group（0.025 ± 0.003, all P 〈 0.05）;compared with 0 mg/L group（0. 115 ± 0.011）, the KBD 0.2500 mg/L dose group promoted cell proliferation（0.128 ± 0.037, P 〈 0.05）, the KBD 1.0000 mg/L dose group inhibited cell growth （0.071 ± 0.019, P 〈 0.05）. The apoptotic rate of 0.0500 - 1.0000 mg/L dose control group [ （18.88 ± 0.02）%,（17.58 ± 0.01）%, （17.09 ± 0.04）%, （56.00 ± 0.02）%, （57.85 ± 0.03）% ] were higher than that of the 0 mg/L group[（13.51 ± 0.01）%, all P 〈 0.05];compared with 0 mg/L group[（25.84 ± 0.02）%],

关 键 词：大骨节病 硒 软骨细胞 细胞凋亡 

分 类 号：R686[医药卫生—骨科学]