血管内皮生长因子对关节软骨细胞诱导型一氧化氮合酶的影响

Influence of vascular endothelial growth factor on inducible nitric oxide synthase expression in rat articular chondrocytes

目的观察血管内皮生长因子（VEGF）对体外培养的关节软骨细胞诱导型一氧化氮合酶（iNOS）表达的影响。方法体外培养SD乳鼠关节软骨细胞，用白细胞介素（IL）-1B诱导的方法建立骨关节炎（OA）体外模型，实验分为4组，每组加入不同处理因素进行干预，A组：（正常对照组）不加任何处理因素；B组：10μg/L VEGF；C组：10μg/L IL-18；D组：10μg／L VEGF＋10μg/L IL-18。采用实时荧光定量PCR（Real TimePCR）检测iNOSmRNA的表达，采用蛋白免疫印迹法（Westernblot）检测iNOS蛋白的表达。结果iNOSmRNA的表达：A组iNOSmRNA无表达，B组（9．64±1．64）、C组（17．27±2．01）及D组（28．93±6．63），3组的iNOSmRNA表达量显著升高，进一步组间比较，D组软骨细胞iNOS的mRNA表达水平明显高于B组（P〈0．01）及C组（P〈0．05），C组软骨细胞iNOS的mRNA表达水平高于B组（P〈0．05）。iNOS蛋白的表达：A组iNOS蛋白无表达，B组（0．44±0．12）、C组（0．74±0．07）及D组（1．38±0．38），3组的iNOS蛋白表达量显著升高，进一步组间比较，D组软骨细胞iNOS的蛋白表达水平明显高于B组（P〈0．01）及C组（P〈0．05），C组软骨细胞iNOS的mRNA表达水平高于B组（P〈0．01）。结论在OA的发病过程中，VEGF可能通过上凋软骨细胞iNOS的表达发挥重要作用。

Objective To investigate the influence of vascular endothelial growth factor （VEGF） on inducible nitric oxide synthase （iNOS） expression in rat articular chondrocytes in vitro. Methods Chondrocytes were isolated and cultured. The model of osteoarthritis （OA） was induced by IL-1β in vitro. Then the experiment was divided into 4 groups ： group A （control） ： without any disposal; group B ： 10μg/L VEGF; group C： 10μg/L IL-1β; group D： 10μg/L VEGF ＋ 10μg/L IL-β The mRNA and protein expression of iNOS was detected by using real-time polymerase chain reaction, and Western blotting respectively. Results The iNOS mRNA expression levels in group B （9. 64± 1.64）, group C （ 17.27 ±2.01） and group D （28.93 ±6. 63） were higher than in group A （control group） significantly, and there was significant difference between groups B and C, B and D, C and D （ B vs C, P 〈0. 05 ; B vs D, P 〈 0. 01 ; C vs D, P 〈 0. 05 ）. As compared with group A, the expression of iNOS protein in group B （0.44 ±0. 12）, group C （0. 74±0.07） and group D （1.38 ±0.38） was significantly increased. There was significant difference between groups B and C, B and D, C and D （ B vs C,P 〈 0.01 ; B vs D,P 〈 0.01 ; C vs D,P 〈 0. 05）. Conclusion VEGF significantly improved the expression of iNOS in rat articular chondrocytes. VEGF plays an important role during the development of OA.