中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2012年
4期
366-370
,共5页
胡龙%王昕%丁轩玺%张超
鬍龍%王昕%丁軒璽%張超
호룡%왕흔%정헌새%장초
血管内皮生长因子类%椎间盘%凋亡%羟脯氨酸%细胞外基质蛋白质类
血管內皮生長因子類%椎間盤%凋亡%羥脯氨痠%細胞外基質蛋白質類
혈관내피생장인자류%추간반%조망%간포안산%세포외기질단백질류
Vascular endothelial growth factors%Intervertebral disc%Apoptosis%Hydroxyproline%Extracellular matrix proteins
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.
目的 觀察血管內皮細胞生長因子(vasctdar endothelial cell growth factor,VEGF)對體外培養大鼠椎間盤細胞及基質代謝的影響. 方法 建立大鼠椎間盤細胞培養體繫,體外單層培養大鼠椎間盤細胞,取生長良好的第2代細胞,使用抗- Fas抗體誘導凋亡,加入不同濃度(10,100,1 000μg/L)的VEGF,影響椎間盤細胞的凋亡及代謝過程.利用流式細胞儀PI標記法檢測椎間盤細胞凋亡情況；利用氯胺-T法和DMB比色法分彆檢測細胞培養液中羥脯氨痠和蛋白多糖的含量. 結果 (1)加入不同濃度VEGF(10,100,1 000μg/L)後,椎間盤細胞凋亡率為(87.62±11.06)％、(53.30 ±9.23)％和(16.75±4.21)％,與對照組比較,差異均有統計學意義(P＜0.01).(2)隨著VEGF濃度的增加(10,100,1 000 μg/L),羥脯氨痠含量[(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]明顯增加.加入不同濃度VEGF後,羥脯氨痠和蛋白多糖的含量與對照組比較,差異均有統計學意義(P＜0.01).羥脯氨痠含量、蛋白多糖含量與VEGF濃度呈正相關( ra=0.972,P＜0.01；rb=0.907,P＜0.01).結論 VEGF可以抑製體外培養的椎間盤細胞凋亡,促進椎間盤細胞基質中膠原及蛋白多糖的閤成.
목적 관찰혈관내피세포생장인자(vasctdar endothelial cell growth factor,VEGF)대체외배양대서추간반세포급기질대사적영향. 방법 건립대서추간반세포배양체계,체외단층배양대서추간반세포,취생장량호적제2대세포,사용항- Fas항체유도조망,가입불동농도(10,100,1 000μg/L)적VEGF,영향추간반세포적조망급대사과정.이용류식세포의PI표기법검측추간반세포조망정황；이용록알-T법화DMB비색법분별검측세포배양액중간포안산화단백다당적함량. 결과 (1)가입불동농도VEGF(10,100,1 000μg/L)후,추간반세포조망솔위(87.62±11.06)％、(53.30 ±9.23)％화(16.75±4.21)％,여대조조비교,차이균유통계학의의(P＜0.01).(2)수착VEGF농도적증가(10,100,1 000 μg/L),간포안산함량[(6.71±0.33) μg/L、(9.12±0.41) μg/L、(11.58±0.12)μg/L]、단백다당함량[(23.21±2.87)μg/L、(32.45±5.23) μg/L、(37.18±3.22) μg/L]명현증가.가입불동농도VEGF후,간포안산화단백다당적함량여대조조비교,차이균유통계학의의(P＜0.01).간포안산함량、단백다당함량여VEGF농도정정상관( ra=0.972,P＜0.01；rb=0.907,P＜0.01).결론 VEGF가이억제체외배양적추간반세포조망,촉진추간반세포기질중효원급단백다당적합성.
Objective 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.
