The Research Center for Bone And Stem Cells, Nanjing Medical University, Nanjing, PR China.
Cytotherapy. 2011 Feb;13(2):156-64. doi: 10.3109/14653249.2010.512633. Epub 2010 Sep 15.
We wanted to determine whether zinc supplementation can inhibit bone marrow-derived mesenchymal stromal cell (MSC) apoptosis and enhance their tissue regenerative potential a in mouse ischemic hindlimb model.
Rat bone marrow cells were cultured and the resulting MSC were passaged for 3-7 generations. The proliferation and apoptosis of MSC was examined by 3-[4,5-dimethyl-2-thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis. The activation of protein kinases B (Akt) was determined by Western blots. Vascular endothelial growth factor (VEGF) levels were examined by enzyme-linked immunosorbent assay. The mouse hindlimb ischemic model was established by ligating the right femoral artery. Mice received MSC, zinc-treated MSC or vehicle. The blood flow was assessed by laser Doppler imaging. The survival rate of donor cells was quantified by real-time polymerase chain reaction for the sex-determining region of the Y-chromosome (Sry). Angiogenesis was assessed by histochemical staining and immunofluoresence staining.
Supplementation with physiologic amounts of zinc caused a marked attenuation of cell apoptosis, enhanced cell viabilities, increased VEGF release and up-regulated Akt activation. Zinc-treated MSC delivered into ischemic hindlimbs resulted in significant improvements in limb blood perfusion by increased implanted MSC survival and stimulated angiogenesis.
This study demonstrates the potential of zinc supplement to enhance survival of engrafted MSC and ameliorate their tissue regenerative potential in a mouse ischemic hindlimb model.
我们旨在确定锌补充剂是否能抑制骨髓间充质干细胞（MSC）凋亡，并增强其在小鼠缺血后肢模型中的组织再生潜能。
培养大鼠骨髓细胞，传代培养 MSC 3-7 代。通过 3-[4,5-二甲基-2-噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）检测和流式细胞术分析检测 MSC 的增殖和凋亡。通过 Western blot 检测蛋白激酶 B（Akt）的激活。通过酶联免疫吸附试验（ELISA）检测血管内皮生长因子（VEGF）水平。通过结扎右股动脉建立小鼠后肢缺血模型。小鼠接受 MSC、锌处理的 MSC 或载体。通过激光多普勒成像评估血流。通过实时聚合酶链反应（PCR）检测性别决定区 Y 染色体（Sry）定量供体细胞的存活率。通过组织化学染色和免疫荧光染色评估血管生成。
补充生理剂量的锌可显著减轻细胞凋亡，增强细胞活力，增加 VEGF 释放，并上调 Akt 激活。将锌处理的 MSC 植入缺血后肢可通过增加植入 MSC 的存活率和刺激血管生成来显著改善肢体血流灌注。
这项研究表明，锌补充剂具有增强植入 MSC 存活和改善其在小鼠缺血后肢模型中组织再生潜能的潜力。
