CAJ | 학술논문

目的观察Caveolin-1( Cav-1)在一氧化氮(N0)介导的血管新生中的作用.方法建立人脐静脉内皮细胞(HUVECs)三维纤维蛋白凝胶血管新生模型.通过管腔形成指数(TFI)来定量分析毛细血管管腔样结构,以亚硝酸盐来反映NO的产生量.根据是否添加血管内皮生长因子(VEGF)和N-硝基-L-精氨酸(L-NAME)分为VEGF组、VEGF+L-NAME组和对照组.HUVECs转染反义寡核苷酸( ASON)和无义寡核苷酸(NSON),并将其分为ASON组、NSON组和对照组.结果 VEGF组TFI和亚硝酸盐分别为(3.15±0.46)、(0.36±0.05) mmol/L,明显高于对照组(0.89±0.09)、(0.18 ±0.02) mmol/L(P＜0.05).VEGF+ L-NAME组TFI和亚硝酸盐分别为(1.06±0.13)、(0.21±0.02) mmol/L,与对照组比较差异无统计学意义(P＞0.05).HUVECs在三维纤维蛋白凝胶内培养后Cav-1的表达水平逐渐升高.ASON组TFI(1.28±0.15)明显低于对照组(3.27 ±0.33)和NSON组(3.05±0.31) (P ＜0.05),转染ASON的HUVECs添加VEGF和L-NAME后TFI和亚硝酸盐无明显改变,差异有统计学意义(P＞0.05).结论 Cav-1对NO介导的血管新生起非常重要的调节作用.
목적 관찰Caveolin-1( Cav-1)재일양화담(N0)개도적혈관신생중적작용.방법 건립인제정맥내피세포(HUVECs)삼유섬유단백응효혈관신생모형.통과관강형성지수(TFI)래정량분석모세혈관관강양결구,이아초산염래반영NO적산생량.근거시부첨가혈관내피생장인자(VEGF)화N-초기-L-정안산(L-NAME)분위VEGF조、VEGF+L-NAME조화대조조.HUVECs전염반의과핵감산( ASON)화무의과핵감산(NSON),병장기분위ASON조、NSON조화대조조.결과 VEGF조TFI화아초산염분별위(3.15±0.46)、(0.36±0.05) mmol/L,명현고우대조조(0.89±0.09)、(0.18 ±0.02) mmol/L(P＜0.05).VEGF+ L-NAME조TFI화아초산염분별위(1.06±0.13)、(0.21±0.02) mmol/L,여대조조비교차이무통계학의의(P＞0.05).HUVECs재삼유섬유단백응효내배양후Cav-1적표체수평축점승고.ASON조TFI(1.28±0.15)명현저우대조조(3.27 ±0.33)화NSON조(3.05±0.31) (P ＜0.05),전염ASON적HUVECs첨가VEGF화L-NAME후TFI화아초산염무명현개변,차이유통계학의의(P＞0.05).결론 Cav-1대NO개도적혈관신생기비상중요적조절작용.
Objective To examine the role of caveolin-1 (Cav-1) in nitric oxide (NO)-mediated angiogenesis.Methods Human umbilical vein endothelial cells (HUVECs) were cultured in three dimensional fibrin gels to form capillary-like tubules.The tubule formation index (TFI) and nitrite were measured to quantify the capillary-like tubes and production of NO.In the presence or absence of vascular endothelial growth factor (VEGF) and NG-nitro-L-arginine methyl ester (L-NAME),HUVECs were separated into the VEGF group,VEGF + L-NAME group and control group.HUVECs were treated with antisense oligonucleotides (A SON) and nonsense oligonucleotides (NSON) and they were separated into the ASON group,NSON group and control group.Results The TFI and nitrite levels in the VEGF group were (3.15 ±0.46) and (0.36 ±0.05) mmol/L respectively,which were obviously higher than those in the control group (0.89 ± 0.09) and (0.18 ± 0.02 ) mmol/L,P ＜ 0.05,but there were no obvious differences between the VEGF+ L-NAME group (1.06 ±0.13) and (0.21 ±0.02) mmol/L and the control group (P＞0.05).Cav-1 levels weres teadily increased in a time-dependent manner.The TFI level in ASON group was 1.28 ±0.15,which was obviously lower than that in control group (3.27 ±0.33) and NSAN group (3.05 ± 0.31,P ＜ 0.05 ).The TFI and nitrite levels in ASON-transduced HUVECs showed no responses to the addition of VEGF and L-NAME.Conclusion Cav-1 was essential for NO-mediated angiogenesis.