摘要
目的 探讨血小板衍生生长因子 (PDGF) BB在成骨细胞 破骨细胞培养体系中对破骨细胞骨吸收功能的影响。方法 体外分离、培养成人成骨细胞和破骨细胞 ,利用复合培养系统使二者生活在同一环境中 ;在复合培养体系中施加不同浓度的PDGF BB或者PDGF BB +L 单甲基精氨酸 (L NMMA) ;酶动力学法测定培养上清中抗酒石酸酸性磷酸酶 (TRAP)的活性 ;利用甲苯胺蓝对骨吸收陷窝染色并在图像分析仪下测定骨吸收陷窝的面积和数目。结果 在PDGF BB单独作用下 ,复合培养体系中TRAP的活性不随POGF BB的浓度发生明显变化 (P >0 .0 5 ) ,骨吸收陷窝的面积和数目PDGF BB均与对照组比较 ,无明显变化 (P >0 0 5 ) ;在加入L 单甲基精氨酸后 ,随着PDGF BB的浓度递增 ,培养上清中TRAP活性从 1.46U/L± 0 .10U/L升高至最高为 2 .47U/L± 0 .38U/L(P <0 .0 1) ;骨吸收陷窝的面积和数目分别从 436 μm2 ± 147μm2 和 (14.1± 1.2 )个 /片增加到最多为 5 92 μm2 ± 171μm2 和(2 2 .5± 1.7)个 /片 (P <0 .0 1)。在同一PDGF BB浓度下 ,加入L 单甲基精氨酸后 ,TRAP和骨吸收陷窝面积、数量比加入L 单甲基精氨酸前均显著升高。所有数据进行拉丁方设计的方差分析。结论 在成骨细胞 破骨细胞复合培养体系中 ,由于PDGF
Objective To investigate the biological effects of platelet derived growth factor (PDGF) BB on the osteoclasts in the osteoblast osteoclast co culture system. Methods The human osteoblasts and osteoclasts were isolated from the iliac crest of patients and co cultured in the same system. The PDGF BB or PDGF BB+NO synthase inhibitor, N(G) monomethyl L arginine (L NMMA) was administered to the co culture system. The tartrate resistant acid phosphatase (TRAP) activities in the medium were measured by enzyme kinetics. The resorption pits were stained by toludine blue. The area and number of the resorption pits were determined with the Leica Quantimet 500 system. Results Under the stimuli of PDGF BB in the co culture system, TRAP activities changed from 1.40 U/L±0.12 U/L to 1.46 U/L±0.08 U/L ( P >0.05) in the medium, and the area and number of resorption pits was changed from (427±149) μm 2 and (13.3±0.9) per slice to (436±147) μm 2 and (14.1±1.2) per slice respectively ( P >0.05). After the L NMMA was applied to the system, the TRAP activities rose significantly from 1.46 U/L±0.10 U/L to 2.47 U/L±0.38 U/L along with the incnease of dose of PDGF BB, and the number and area of the resorption pits increased significantly from 436 μm 2±147 μm 2 and (14.1±1.2) per slice to 592 μm 2±171 μm 2 and (22 5±1.7) per slice. Both the TRAP activities and resorption pits rose significantly after the L NMMA was administered. Comparisons of different treatment groups were made using analysis of variance (ANOVA) with multiple comparisons and a Student Newman Keuls test. Conclusions The nitric oxide (NO) was produced by osteoblasts stimulated by PDGF BB in the osteoblast osteoclast co culture system. Therefore, the direct promotion effects of PDGF BB on the osteoclasts were inhibited.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2001年第7期425-428,共4页
National Medical Journal of China
基金
默沙东国际交流基金
