摘要
目的:研究VEGF诱导血管内皮细胞产生细胞外H2O2及其在VEGF促血管内皮细胞增殖功能中的作用。方法:①以H2DCFDA为H2O2指示剂,检测500μg/LVEGF刺激下,血管内皮细胞H2O2的产生;②以MTT法检测3×106U/L过氧化氢酶(CAT),及外源性加入5-20mmol/LH2O2对VEGF促增殖功能的影响。结果:①在VEGF刺激血管内皮细胞15min后,细胞内开始出现逐渐增强的荧光,且随时间逐渐增强,至45min左右最强,随后逐渐减弱;而同时加入过氧化氢酶组的细胞则仅有微弱荧光产生,且荧光强度不随时间变化;②3×106U/L过氧化氢酶对VEGF的促增殖功能有明显的抑制作用(P<0.01);③外源性加入5-10mmol/LH2O2时对血管内皮细胞有明显促增殖作用(P<0.01),但其对VEGF的促增殖功能却有显著抑制作用(P<0.01)。结论:VEGF可刺激血管内皮细胞产生细胞外H2O2,在促细胞增殖中可能具有重要作用。而外源性H2O2对VEGF的生理功能可能具有抑制作用。
AIM: To study the effect of VEGF on extracellular H2O2 production in HUVECs and the role of H2O2 in the VEGF- induced proliferation. METHODS: HUVECs was stimulated with 500 ug/L VEGF. Products of extraeellular H2O2 was detected by H2DCFDA staining. MTT method was used to value the influences of 3 × 10^6 U/L eatalase and 5 -20 mmol/L H2O2 to VEGF function. RESULTS: After treatment for 15 min with VEGF, HUVECs appeared fluorescence, and continued to become stronger, peaked at 45 min then decreased. HUVECs, which was treated simultaneity with VEGF and 3 × 10^6 U/L eatalase, only appeared very faint fluorescence. The proliferation of HUVECs by VEGF was restrained when treated with 3 × 10^6 U/L eatalase. The extrinsic H2O2 at concentration of 5 - 10 mmol/L promoted the proliferation of HUVECs but inhibited the proliferation effect of VEGF on HUVECs ( P 〈 0. 01 ). CONCLUSION : These findings indicate that VEGF induces HUVECs to produce extraeellular H2O2 and plays role in proliferation, but extrinsic H2O2 restrains VEGF function.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第3期533-535,共3页
Chinese Journal of Pathophysiology
基金
国家自然科学重点基金资助项目(No30230305)
国家自然科学青年基金资助项目(No30500466)