低氧对骨髓间质细胞表达VEGF的影响

目的:研究低氧对骨髓间质细胞(MSCs)分泌VEGF及促血管内皮细胞(VECs)增殖的影响。方法:用VEGF ELISA试剂盒和Micro BCA蛋白质定量试剂测定正常氧和低氧培养的MSCs的上清液中VEGF与总蛋白的含量的比值(pg·μg-1)。用不同成份配制的1%FBS的DMEM培养基培养VECs,实验分成5组:低氧MSCs组(低氧培养的MSCs的上清液)、VEGF组(与低氧MSCs组等浓度VEGF)、antibody-低氧MSCs组(低氧MSCs组中加入10μg·ml-1anti-VEGF)、antibody-VEGF组(VEGF组中加入10μg·ml-1anti-VEGF)和对照组(不加其它成份)。用MTT法检测VECs的增殖程度。结果:在低氧条件下培养的MSCs分泌的VEGF明显高于正常氧条件下培养的MSCs分泌的VEGF(P<0.01)。促进VECs增殖的程度依次为低氧MSCs组>VEGF组>antibod-y-低氧MSCs组>antibody-VEGF组=control组。结论:MSCs在低氧条件下比在正常氧条件下具有更强的分泌VEGF的能力。MSCs可分泌VEGF和其它细胞因子共同促进VECs增殖,但VEGF发挥主要作用。

Effect of Hypoxia on VEGF Expressing of Bone Marrow Stromal Cells

Objective: To study the effect of hypoxia on bone marrow stromal cells ’(MSCs) ability to express VEGF and promote the proliferation of vascular endo thelial cells(VECs). Methods: The supernatant of rabbit MSCs wh ich were cultured under normoxic or hypoxic conditions was collected from the cu ltured media. The concentration of VEGF in the supernatant was measured by using sandwich ELISA kits and was corrected for total cell protein by Micro BCA prote in assay. VECs were cultured in five groups of 1% FBS DMEM media as follows: gr oup of hypoxic MSCs (the supernatant of hypoxic MSCs), goup of VEGF (add VEGF t o the same concentration with that in the group of hypoxic MSCs), group of antib odyMSCs (add 10 μg·ml~-1 antiVEGF antibody to the group of hypoxic MS Cs), group of antibodyVEGF (add 10 μg·ml~-1 antiVEGF antibody to the group of VEGF), and the group of control (add nothing). The proliferation of the VECs was measured by MTT assay. Results: The MSCs which cultur ed under hypoxic conditions secreted more VEGF than the MSCs under normoxic cond itions did (P<0.01). The ability to promote VECs’ proliferation ranked from stong to weak as follows: hypoxic MSCs>VEGF>antibody hypoxic MSCs>ant ibodyVEGF=control. Conclusion: The hypoxic MSCs’ ability to screte VEGF is higer than the normoxic MSCs’. MSCs can secret VEGF and other c ytokines which promotes VECs’ proliferation together and VEGF take the major ro le.