论文部分内容阅读
背景:研究表明,地塞米松可以使关节软骨表层基质中的Ⅱ型胶原减少,Ⅰ型胶原增多,但糖皮质激素对关节软骨细胞增殖作用的机制尚不十分清楚。目的:验证地塞米松磷酸钠对体外培养兔关节软骨细胞增殖的影响。设计:对比观察。材料:1月龄新西兰白兔用于软骨细胞的分离与培养。方法:兔关节软骨细胞常规培养后随机分为对照组和实验组,对照组用不含地塞米松磷酸钠的DMEM培养液培养,实验组则分别加入含不同质量浓度地塞米松磷酸钠(0.02,0.1,0.5g/L)的DMEM培养液。主要观察指标:①原代及传代兔关节软骨细胞的形态学观察。②应用流式细胞术及免疫细胞化学法检测软骨细胞增殖及其合成Ⅱ型胶原能力。③透射电子显微镜观察软骨细胞超微结构的变化。结果:实验组软骨细胞贴壁、增殖较对照组缓慢。各实验组Ⅱ型胶原平均灰度值均显著高于对照组(P<0.05)。实验组软骨细胞进入S期、G2+M期的细胞比率较对照组软骨细胞减少、进入G0/G1期的细胞比率增加。电镜下见软骨细胞线粒体、粗面内质网数量减少。结论:地塞米松磷酸钠抑制软骨细胞的增殖,可能与抑制关节软骨细胞Ⅱ型胶原和蛋白的合成能力有关。
BACKGROUND: Dexamethasone can reduce type Ⅱ collagen and type Ⅰ collagen in the surface matrix of articular cartilage, but the mechanism of glucocorticoid on the proliferation of articular chondrocytes is not yet clear. Objective: To verify the effect of dexamethasone sodium phosphate on rabbit articular chondrocytes proliferation in vitro. Design: comparative observation. MATERIALS: One-month old New Zealand white rabbits were used for the isolation and culture of chondrocytes. Methods: Rabbit articular chondrocytes were randomly divided into control group and experimental group. The control group was cultured in DMEM without dexamethasone sodium phosphate. The experimental group was treated with different concentrations of dexamethasone sodium phosphate (0.02 , 0.1, 0.5 g / L) DMEM culture medium. MAIN OUTCOME MEASURES: ① Morphological observation of primary and passage rabbit articular chondrocytes. ② Flow cytometry and immunocytochemistry were used to detect chondrocyte proliferation and its ability to synthesize type Ⅱ collagen. Transmission electron microscopy was used to observe the change of ultrastructure of chondrocytes. Results: The experimental group chondrocytes adherent, proliferation slower than the control group. The average gray value of type Ⅱ collagen in experimental group was significantly higher than that in control group (P <0.05). In the experimental group, the percentage of chondrocytes entering the S phase and G2 + M phase decreased compared with that of the control group, and the percentage of cells entering the G0 / G1 phase increased. Electron microscope, chondrocytes mitochondria, the number of rough endoplasmic reticulum decreased. Conclusion: Dexamethasone sodium phosphate inhibits the proliferation of chondrocytes, which may be related to the inhibition of the synthesis of type Ⅱ collagen and protein in articular chondrocytes.