High glucose conditions suppress the function of bone marrow-derived endothelial progenitor cells via inhibition of the eNOS-caveolin-1 complex
High glucose conditions suppress the function of bone marrow-derived endothelial progenitor cells via inhibition of the eNOS-caveolin-1 complex
复制标题
高葡萄糖条件通过抑制 eNOS-caveolin-1 复合物抑制骨髓源性内皮祖细胞的功能
DOI：
10.3892/mmr.2011.644
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发表时间：
2012-02-01
影响因子：
3.4
通讯作者：
Zhang, Nan
中科院分区：
文献类型：
--
作者：
Cao, Chunli;Zhang, Hongkun;Zhang, Nan
本研究旨在揭示高糖对体外培养的大鼠骨髓内皮祖细胞的影响。取骨髓总单个核细胞，在内皮细胞生长介质-2中培养，荧光显微镜下鉴定。免疫荧光法鉴定内皮祖细胞表达VEGFR-2和CD133，进一步鉴定为Dil-Ac-LDL摄取和FITC-UEA-1凝集素结合双阳性的贴壁细胞。收集贴壁细胞，加入不同终浓度的葡萄糖(11.1、33.3、55.5 mmol/L)。检测内皮祖细胞的增殖、迁移和体外血管生成能力。检测细胞小窝蛋白-1和内皮型一氧化氮合酶(ENOS)基因和蛋白水平的变化，并检测细胞培养上清液中一氧化氮(NO)的含量。结果表明，随着培养液中葡萄糖浓度的增加，细胞的增殖、迁移和体外血管生成能力逐渐降低，而小窝蛋白-1基因的mRNA和蛋白水平逐渐升高。内皮型一氧化氮合酶基因的表达水平无明显变化，但蛋白表达减少，培养液中NO水平下降。综上所述，高糖环境对体外培养的骨髓源性内皮祖细胞的功能是不利的，可能是通过破坏eNOS-小窝蛋白-1复合体，导致NO合成减少，最终导致内皮祖细胞功能受损。
The present study aimed to reveal how high glucose affects rat bone marrow-derived endothelial progenitor cells in vitro. Total mononuclear cells of bone marrow were obtained, cultured in endothelial cell growth medium-2 and identified by fluorescence microscopy. Using immunofluorescence, endothelial progenitor cells were identified by expression of VEGFR-2 as well as CD133 and were further characterized as those adherent cells which were double positive by Dil-Ac-LDL uptake and FITC-UEA-1 lectin binding. The attached cells were collected and glucose was added to the culture medium at various final concentrations (11.1, 33.3 and 55.5 mmol/l). Proliferation, migration and in vitro angiogenic ability of endothelial progenitor cells were measured. The change in mRNA and protein levels of caveolin-1 and endothelial nitric oxide synthase (eNOS) were examined; in addition, nitric oxide (NO) levels in the cell medium were measured. Based on the results, with increasing glucose concentration in the medium, proliferation, migration and in vitro angiogenic capacity of the cells were reduced, whereas mRNA and protein levels of caveolin-1 gene increased gradually. The mRNA levels of the eNOS gene did not differ, but the protein expression was reduced and NO levels in the culture medium declined. In conclusion, high glucose conditions are detrimental to the function of bone marrow-derived endothelial progenitor cells in vitro, probably by damaging the eNOS-caveolin-1 complex, which results in the reduction of NO synthesis, eventually leading to the impaired function of endothelial progenitor cells.