血管内皮生长因子对关节软骨细胞Ⅱ型胶原表达的影响

Effect of vascular endothelial growth factor on collagen Ⅱ expression in rat articular chondrocytes

目的观察血管内皮生长因子（VEGF）对关节软骨细胞Ⅱ型胶原表达的影响。方法体外培养乳鼠关节软骨细胞，实验分为4组，加入不同因素干预。A组（对照组）：不加处理因素；B组：10ng／mlVEGF；C组：10ng／ml白细胞介素（IL）-1β；D组：10ng／mlVEGF＋10ng／mlIL-1β。采用实时荧光定量聚合酶链反应检测Ⅱ型胶原mRNA的表达，采用蛋白免疫印迹法检测Ⅱ型胶原蛋白的表达。采用单因素方差分析进行统计学处理。结果各组软骨细胞Ⅱ型胶原mRNA的相对表达量分别为：A组1．00±0．08，B组0．78±0．07，C组0．67±0．06，D组0．57±0．04，B、C及D组软骨细胞Ⅱ型胶原的mRNA表达量均显著低于A组（P均〈0．01），D组Ⅱ型胶原的mRNA表达水平明显低于B组及C组；与A组（0．95±0．21）比较，B组（0．71±0．14）、C组（0．60±0．11）及D组（0．31±0．09）的Ⅱ型胶原蛋白表达量显著降低（P均〈0．01），D组软骨细胞Ⅱ型胶原的蛋白表达水平明显低于B组及C组（P均〈0．01）。结论在骨关节炎的发病过程中，VEGF可能通过抑制关节软骨细胞Ⅱ型胶原的表达发挥重要作用。

Objective To investigate the effect of vascular endothelial growth factor （VEGF） on colla- gen Ⅱexpression in rat articular chondrocytes in vitro. Methods Chondrocytes were isolated and cultured. Then rats were divided into 4 groups： group A （control）： without any intervention; group B： 10 ng/ml VEGF was added; group C： 10 ng/ml IL-1β was added; group D： 10 ng/ml VEGF and 10 ng/ml IL-1β were added. Messenger RNA （mRNA） expression of collagen Ⅱ was detected by using real time polymerase chain reaction （real time PCR）, and the protein expression level of collagen H was detected by Western blotting. Comparisons between groups were performed by one-way ANOVA Results The collagen Ⅱ mRNA expression levels of group B （0.78±0.07）, group C （0.67＋0.06） and group D （0.57＋0.04） were significantly lower than those of the group A （1.00±0.08）, and there was significant difference between B and D, C and D. Compared with group A （0.95±0.21）, the expression of collagen Ⅱ protein in group B （0.71±0.14）, group C （0.60±0.11） and group D （0.31±0.09） was significantly suppressed. The expression of collagen Ⅱprotein in group D was significantly lower than those of group B and C. Conclusion VEGF can significantly suppress the expression of collagen Ⅱ in rat articular chondrocytes. VEGF may play an important role in the development of osteoarthritis.