基质细胞衍生因子-1对兔膝关节软骨细胞增殖和凋亡的影响  

Effects of stromal cell-derived factor-1 on the proliferation and apoptosis of articular chondrocytes in rabbits

作　　者：王世东[1] 王宇泽[1] 曾令员[1] 贺冬冬[1] 王春芳[1] 卫小春[1] 

机构地区：[1]山西医科大学第二医院骨科,太原030001

出　　处：《中华实验外科杂志》2013年第4期781-783,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（31200728、81071495）

摘　　要：目的观察基质细胞衍生因子-1（SDF-1）对兔膝关节软骨细胞体外增殖的影响，探讨SDF—l在软骨细胞凋亡过程中的作用。方法体外分离培养兔膝关节软骨细胞并随机分为4组：对照组、50μg／LSDF．1组、50μ/LSDF-1＋5mg／LCXC趋化因子受体4特异性拮抗剂（AMD3100）作用组、5mg／LAMD3100处理组，采用噻唑蓝（MTT）比色法和测定细胞周期观察各组软骨细胞增殖。同时将各组置于250μmol／L过氧化氢条件下作用2h，锥虫蓝染色计数软骨细胞存活率；测定各组中半胱氨酰天冬氨酸特异性蛋白酶-3（Caspase．3）的活力；流式细胞仪观察各组软骨细胞凋亡率的变化；原位末端转移酶标记（TUNEL）法检测凋亡的软骨细胞。结果增殖实验中SDF-1组吸光度（A）值（0．504±0．024）均高于其他各组，且该组S＋G2／M期细胞比例高于其他各组，差异有统计学意义（p〈0．05）、在凋亡方面，锥虫蓝拒染实验结果显示SDF-1组中细胞存活率高于其他3组（P〈0．01）：而SDF-1组膜联蛋白V-异硫氰酸荧光素（AnnexinV—FITC）标记的细胞比例[（4．00±0.92）,低于其他各组（P〈0．05）；SDF-1组Caspase-3活力（40．03±2．56）nmolpNA／（h·rag）蛋白也较其他组低（p〈0．01）：TUNEL实验中SDF-1组细胞核呈棕黄色的软骨细胞数低于其他各组（P〈0．05）．但各实验中SDF—l＋AMD3100组和AMD3100处理组与对照组比较，差异无统计学意义（P〉0．05）。结论SDF-1可促进兔膝关节软骨细胞体外增殖，且在50μg／L时软骨细胞增殖活性好，同时SDF－1在过氧化氯诱导的细胞凋亡条件下可保护软骨细胞免于凋亡。Objective To observe the effect of stromal cell-derived factor-1 （SDF-1） on the pro- liferation of articular chondroeytes in rabbits and discuss the role of SDF-1 in ehondrocyte apoptosis. Meth- ods The chondrocytes were isolated and eutured, then divided into 4 groups： control group, 50 μg/L SDF-1 group, 50 μg/L SDF-1 ＋ 5 mg/L CXC chemokine receptor-4 specific antagonist （AMD3100） group, and 5 mg/L AMD3100 group. The MTT assay and methods for cell cycle were used to observe the proliferation of chondroeytes. In addition, each group was exposured to the condition of 250 μmol/L hydro- gen peroxide. Then cell viability was detected by using trypan blue exclusion assay, apoptosis rate of chon- drocytes was detected by using flow cytometry, and the dynamic changes of Caspase-3 were measured in each group. Furthermore, the apoptosis of chondrocytes was evaluated by TdT-mediated dUTP nick end la- beling （TUNEL） assay. Results The absorbanee value of SDF-1 set （0. 504 ＋ 0. 024） was higher than oth- er groups, and the S ＋ G2/M cell ratio in SDF-1 group was higher than other groups with the differnce bing statistically significant （ all P 〈 0. 05）. In terms of apoptosis, the viable rate in SDF-1 group was higher than other groups （P 〈0. 01 ）, while the Annexin V-FITC（ ＋ ） proportion of cartilage cells in the SDF-1 group （4. O0 ＋0.92）% was lower than other groups （P 〈0. 05）. Caspase-3 activity in SDF-1 group [ （40.03 ＋ 2. 56） nmolpNA/（h.mg） protein] was lower than other groups （P 〈0. 01 ）, and in addition, the number of apoptotic cells in SDF-1 group was less than other groups （ P 〈 0. 05 ）, but there was no significant difference among control group, SDF-1 ＋ AMD3100 group and AMD3100 group in all experiments. Conclu- sion SDF-1 could contribute to the proliferation of rabbit chondrocytes in vitro, and in 50 μg/L group the proliferation activity is reasonable, meanwhile SDF-1 can protect the chondrocytes from apoptosis induced by hydrogen per

关 键 词：基质细胞衍生因子-1 脱噬作用 增殖 软骨细胞 兔 

分 类 号：R6[医药卫生—外科学]