Generation of an anti-angiogenic endothelial progenitor cell line via endostatin gene transfer.
Generation of an anti-angiogenic endothelial progenitor cell line via endostatin gene transfer.
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通过内皮抑素基因转移产生抗血管生成内皮祖细胞系
DOI：
10.3892/mmr.2018.8623
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发表时间：
2018-04
影响因子：
3.4
通讯作者：
Yao K
中科院分区：
文献类型：
--
作者：
Ai J;Sun JH;Wan T;Ma J;Feng L;Yao K
本研究对内皮祖细胞 (EPC) 作为新生血管形成 (NV) 治疗方法的可行性进行了调查。此外，内皮抑素先前已被证明是血管生成的抑制剂和血管渗漏的抑制剂。本研究的目的是产生具有抗血管生成作用的转基因 EPC，用于治疗眼部 NV。从大鼠外周血样本中获取EPC并进行验证。生成慢病毒-内皮抑素-绿色荧光蛋白重组构建体并用于感染 EPC。然后对转染的细胞进行嘌呤霉素选择。然后应用逆转录定量聚合酶链反应和蛋白质印迹测定来分别测定内皮抑素mRNA和蛋白质表达水平。此外，还检测血管内皮生长因子（VEGF）的表达水平，以观察内皮抑素转染的EPC的抗血管生成作用。嘌呤霉素 (1 µg/ml) 选择 4 天后，生成稳定的内皮抑素转染 EPC 系。在这个稳定的内皮抑素转染的EPC系中，内皮抑素的表达水平增加；而VEGF的表达水平下降。本研究的结果表明，EPCs可以通过基因修饰来过度表达内皮抑素，这可能通过增加内皮抑素的表达和减少VEGF的表达来为细胞提供抗血管生成作用。因此，经过基因修饰以过表达内皮抑素的 EPC 可以作为眼部 NV 治疗的潜在治疗剂。
The viability of endothelial progenitor cells (EPCs) as a therapeutic treatment for neovascularization (NV) was subject to investigation in the present study. Furthermore, endostatin has previously been demonstrated to be an inhibitor of angiogenesis and a suppressant of vascular leakage. The aim of the present study was to generate transgenic EPCs with anti-angiogenic effects for the treatment of ocular NV. EPCs were obtained from rat peripheral blood samples and then verified. A lentiviral-endostatin-green fluorescent protein recombinant construct was generated and used to infect EPCs. Transfected cells were then subjected to puromycin selection. Reverse transcription-quantitative polymerase chain reaction and a western blot assay were then applied in order to determine both the endostatin mRNA and protein expression levels, respectively. In addition, vascular endothelial growth factor (VEGF) expression levels were also detected in order to observe the anti-angiogenic effect of the endostatin-transfected EPCs. Following puromycin (1 µg/ml) selection for 4 days, a stable endostatin-transfected EPC line was generated. In this stable endostatin-transfected EPC line, the expression levels of endostatin increased; whereas the expression levels of VEGF decreased. The results of the present study revealed that EPCs can be genetically modified to overexpress endostatin, which may provide the cells with an anti-angiogenic effect via increased expression of endostatin and decreased expression of VEGF. Thus, EPCs genetically modified to overexpress endostatin may serve as a potential therapeutic agent for ocular NV treatment.
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影响因子：
15.9
作者：
Butler, JM;Guthrie, SM;Scott, EW
通讯作者：
Scott, EW
影响因子：
56.9
作者：
Asahara, T;Murohara, T;Isner, JM
通讯作者：
Isner, JM
DOI：
10.2147/opth.s27423
发表时间：
2012
期刊：
Clinical ophthalmology (Auckland, N.Z.)
影响因子：
--
作者：
Baharivand N;Zarghami N;Panahi F;Dokht Ghafari MY;Mahdavi Fard A;Mohajeri A
通讯作者：
Mohajeri A
影响因子：
7.7
作者：
Caballero, Sergio;Sengupta, Nilanjana;Grant, Maria B.
通讯作者：
Grant, Maria B.
影响因子：
2.3
作者：
Bai, Yu-jing;Huang, Lv-zhen;Li, Xiao-xin
通讯作者：
Li, Xiao-xin