摘要目的 观察血管内皮细胞生长因子(vasctdar endothelial cell growth factor,VEGF)对体外培养大鼠椎间盘细胞及基质代谢的影响. 方法 建立大鼠椎间盘细胞培养体系,体外单层培养大鼠椎间盘细胞,取生长良好的第2代细胞,使用抗- Fas抗体诱导凋亡,加入不同浓度(10,100,1 000μg/L)的VEGF,影响椎间盘细胞的凋亡及代谢过程.利用流式细胞仪PI标记法检测椎间盘细胞凋亡情况；利用氯胺-T法和DMB比色法分别检测细胞培养液中羟脯氨酸和蛋白多糖的含量. 结果 (1)加入不同浓度VEGF(10,100,1 000μg/L)后,椎间盘细胞凋亡率为(87.62±11.06)％、(53.30 ±9.23)％和(16.75±4.21)％,与对照组比较,差异均有统计学意义(P＜0.01).(2)随着VEGF浓度的增加(10,100,1 000 μg/L),羟脯氨酸含量[(6.71±0.33) μg/L、(9.12±0.41) μg/L、(11.58±0.12)μg/L]、蛋白多糖含量[(23.21±2.87)μg/L、(32.45±5.23) μg/L、(37.18±3.22) μg/L]明显增加.加入不同浓度VEGF后,羟脯氨酸和蛋白多糖的含量与对照组比较,差异均有统计学意义(P＜0.01).羟脯氨酸含量、蛋白多糖含量与VEGF浓度呈正相关( ra=0.972,P＜0.01；rb=0.907,P＜0.01).结论 VEGF可以抑制体外培养的椎间盘细胞凋亡,促进椎间盘细胞基质中胶原及蛋白多糖的合成.

abstractsObjective To observe the effect of vascular endothelial cell growth factor (VEGF)on apoptosis and matrix metabolism of rat intervertebral.disc cells cultured in vitro. Methods Culture system of rat intervertebral disc cells was established to culture the monolayer intervertebral disc cells in vitro.The well-grown intervertebral disc cells of the second generation were chosen and anti-Fas antibody was applied to induce their apoptosis.Then,VEGF at different concentrations ( 10,100,1 000 μg/L)were used to affect their apoptosis and metabolism.The apoptosis of the cells stained with Propidium Iodide (PI) was detected by using flow cytometry.The levels of hydroxyproline and proteoglycan in the culture medium were detected by using chloramines T method and DMB chromometry method,respectively.Results ( 1 ) The apoptotic ratio of the intervertebral disc cells affected by different concentrations of VEGF (10,100,1 000 μg/L) was (87.62 ±11.06)％,(53.30 ±9.23)％ and (16.75 ±4.21)％ respectively,with significant differences in comparison with the control group (P ＜ 0.01 ).(2)The contents of hydroxyproline [ (6.71 ±0.33) μg/L,(9.12 ±0.41 ) μg/L,( 11.58 ±0.12) μg/L] and proteoglycan [ (23.21 ± 2.87) μg/L,( 32.45 ± 5.23 ) μg/L,(37.18 ± 3.22) μg/] in the culture medium raised with the increase of VEGF concentration ( 10,100,1 000 μg/L),with statistical differences compared with those of the control group (P ＜0.01 ).The contents of hydroxyproline and proteoglycan were positively correlated with the concentration of VEGF (ra=0.972,P＜0.01;rb =0.907,P＜0.01). Conclusion VEGF can inhibit the apoptosis of the rat intervertebral disc cells cultured in vitro and promote collagen and proteoglycan synthesis of the cell matrix.