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核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响

刘荣强 袁泽南 吴小材 刘炜 汪国营

刘荣强, 袁泽南, 吴小材, 等. 核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响[J]. 器官移植, 2018, 9(2): 110-115. doi: 10.3969/j.issn.1674-7445.2018.02.004
引用本文: 刘荣强, 袁泽南, 吴小材, 等. 核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响[J]. 器官移植, 2018, 9(2): 110-115. doi: 10.3969/j.issn.1674-7445.2018.02.004
Liu Rongqiang, Yuan Zenan, Wu Xiaocai, et al. Effect of Nrf2 overexpression on anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 110-115. doi: 10.3969/j.issn.1674-7445.2018.02.004
Citation: Liu Rongqiang, Yuan Zenan, Wu Xiaocai, et al. Effect of Nrf2 overexpression on anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 110-115. doi: 10.3969/j.issn.1674-7445.2018.02.004

核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响

doi: 10.3969/j.issn.1674-7445.2018.02.004
基金项目: 

国家十二五科技重大专项 2012ZX10002017-005

国家十二五科技重大专项 2012ZX10002016-023

广东省科技计划项目 2014B020228003

广东省科技计划项目 2014A020211015

广东省自然科学基金 2015A030312013

广东省自然科学基金 2015A030313038

广州市科技计划项目 201400000001-3

广州市科技计划项目 158100076

广州市科技计划项目 2014J4100183

详细信息
    作者简介:

    刘荣强,1990年生,硕士研究生,研究方向为干细胞和肝癌,Email: 425475531@qq.com

    通讯作者:

    汪国营,副教授,博士研究生导师,研究方向为干细胞和肝癌,Email: wanggy3@126.com

  • 中图分类号: R617

Effect of Nrf2 overexpression on anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells

More Information
  • 摘要:   目的  研究核因子E2相关因子2(Nrf2)对间充质干细胞(MSCs)的抗缺氧及抗凋亡能力的影响。  方法  将过表达Nrf2的重组质粒及辅助质粒转染人胚肾细胞(293FT),获得过表达Nrf2的高滴度慢病毒。MSCs分别转染过表达Nrf2慢病毒以及空载体慢病毒,构建稳定过表达Nrf2的Nrf2-MSCs(Nrf2过表达组)以及绿色荧光蛋白(GFP)-MSCs(对照组)。采用荧光显微镜观察两组细胞的绿色荧光表达情况。采用蛋白免疫印迹(Western Blot)法检测两组细胞中Nrf2蛋白的表达水平。采用光学显微镜观察两组细胞的抗缺氧能力,采用流式细胞术检测两组细胞的抗凋亡能力。  结果  成功构建了稳定过表达Nrf2的Nrf2-MSCs,Western Blot法检测结果显示Nrf2过表达组细胞的Nrf2蛋白表达水平明显高于对照组(P < 0.01)。缺氧处理15 h后,Nrf2过表达组的细胞活力明显高于对照组。流式细胞术检测表明Nrf2过表达组细胞的凋亡率为(30.9±1.4)%,明显低于对照组细胞的(61.3±1.3)%(P < 0.05)。  结论  稳定过表达Nrf2的Nrf2-MSCs在低氧环境中具有一定的抗缺氧、抗凋亡能力。

     

  • 图  1  重组质粒阳性克隆菌液的PCR产物

    Figure  1.  PCR products of recombinant plasmid positive clone bacteria solution

    图  2  荧光显微镜下两组细胞的绿色荧光表达(×200)

    A图为Nrf2过表达组细胞;B图为对照组细胞

    Figure  2.  The green fluorescent expression of cells in two groups by fluorescence microscope

    图  3  两组细胞Nrf2蛋白的表达水平比较

    与对照组比较,aP<0.05

    Figure  3.  Comparison of the expression level of Nrf2 protein of cells between two groups

    图  4  两组细胞抗缺氧能力的比较(×100)

    A图为Nrf2过表达组细胞;B图为对照组细胞

    Figure  4.  Comparison of the anti-hypoxia ability of cells between two groups

    图  5  两组细胞抗凋亡能力的比较

    A图为Nrf2过表达组细胞;B图为对照组细胞

    Figure  5.  Comparison of the anti-apoptotic ability of cells between two groups

  • [1] LEAL AJ, TANNURI AC, BELON AR, et al. Effects of ischemic preconditioning in a pig model of large-for-size liver transplantation[J]. Clinics (Sao Paulo), 2015, 70(2): 126-135. DOI: 10.6061/clinics/2015(02)10.
    [2] BEHRENDS M, HIROSE R, SERKOVA NJ, et al. Mild hypothermia reduces the inflammatory response and hepatic ischemia/reperfusion injury in rats[J]. Liver Int, 2006, 26(6): 734-741. doi: 10.1111/liv.2006.26.issue-6
    [3] ICHIHARA S, YAMADA Y, LIU F, et al. Ablation of the transcription factor Nrf2 promotes ischemia-induced neovascularization by enhancing the inflammatory response[J]. Arterioscler Thromb Vasc Biol, 2010, 30(8):1553-1561. DOI: 10.1161/ATVBAHA.110.204123.
    [4] KODE A, RAJENDRASOZHAN S, CAITO S, et al. Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells[J]. Am J Physiol Lung Cell Mol Physiol, 2008, 294(3): L478-L488. doi: 10.1152/ajplung.00361.2007
    [5] MORITO N, YOH K, ITOH K, et al. Nrf2 regulates the sensitivity of death receptor signals by affecting intracellular glutathione levels[J]. Oncogene, 2003, 22(58): 9275-9281. doi: 10.1038/sj.onc.1207024
    [6] KUDOH K, UCHINAMI H, YOSHIOKA M, et al. Nrf2 activation protects the liver from ischemia/reperfusion injury in mice[J]. Ann Surg, 2014, 260(1): 118-127. DOI: 10.1097/SLA.0000000000000287.
    [7] HORWITZ EM, DOMINICI M. How do mesenchymal stromal cells exert their therapeutic benefit?[J]. Cytotherapy, 2008, 10(8): 771-774. DOI: 10.1080/14653240802618085.
    [8] MURPHY MB, MONCIVAIS K, CAPLAN AI. Mesenchymal stem cells: environmentally responsive therapeutics for regenerative medicine[J]. Exp Mol Med, 2013, 45:e54. DOI: 10.1038/emm.2013.94.
    [9] BIANCHI F, SALA E, DONADEI C, et al. Potential advantages of acute kidney injury management by mesenchymal stem cells[J]. World J Stem Cells, 2014, 6(5): 644-650. DOI: 10.4252/wjsc.v6.i5.644.
    [10] MORIGI M, INTRONA M, IMBERTI B, et al. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice[J]. Stem Cells, 2008, 26(8): 2075-2082. DOI: 10.1634/stemcells.2007-0795.
    [11] JIANG MH, LI G, LIU J, et al. Nestin(+) kidney resident mesenchymal stem cells for the treatment of acute kidney ischemia injury[J]. Biomaterials, 2015, 50: 56-66. DOI: 10.1016/j.biomaterials.2015.01.029.
    [12] LINTERMAN KS, PALMER DN, KAY GW, et al. Lentiviral-mediated gene transfer to the sheep brain: implications for gene therapy in batten disease[J]. Hum Gene Ther, 2011, 22(8): 1011-1020. DOI: 10.1089/hum.2011.026.
    [13] KAFRI T, VAN PRAAG H, GAGE FH, et al. Lentiviral vectors: regulated gene expression[J]. Mol Ther, 2000, 1(6): 516-521. doi: 10.1006/mthe.2000.0083
    [14] DREYER JL. Lentiviral vector-mediated gene transfer and RNA silencing technology in neuronal dysfunctions[J]. Mol Biotechnol, 2011, 47(2): 169-187. DOI: 10.1007/s12033-010-9334-x.
    [15] LASEK AW, AZOUAOU N. Virus-delivered RNA interference in mouse brain to study addiction-related behaviors[J]. Methods Mol Biol, 2010, 602: 283-298. DOI: 10.1007/978-1-60761-058-8_17.
    [16] RUBINSON DA, DILLON CP, KWIATKOWSKI AV, et al. A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference[J]. Nat Genet, 2003, 33(3): 401-406. doi: 10.1038/ng1117
    [17] SOHNI A, VERFAILLIE CM. Mesenchymal stem cells migration homing and tracking[J]. Stem Cells Int, 2013: 130763. DOI: 10.1155/2013/130763.
    [18] MOUSA HSE, SHALABY SM, GOUDA ZA, et al. Efficacy of human umbilical cord derived-mesenchymal stem cells in treatment of rat bone marrow exposed to gamma irradiation[J]. Ann Anat, 2017, 210: 64-75. DOI: 10.1016/j.aanat.2016.12.002.
    [19] ZHANG B, SHEN L, SHI H, et al. Exosomes from human umbilical cord mesenchymal stem cells: identification, purification, and biological characteristics[J]. Stem Cells Int, 2016: 1929536. DOI: 10.1155/2016/1929536.
    [20] MA S, XIE N, LI W, et al. Immunobiology of mesenchymal stem cells[J]. Cell Death Differ, 2014, 21(2): 216-225. DOI: 10.1038/cdd.2013.158.
    [21] HOOGDUIJN MJ. Are mesenchymal stromal cells immune cells?[J]. Arthritis Res Ther, 2015, 17:88. DOI: 10.1186/s13075-015-0596-3.
    [22] D'SOUZA N, ROSSIGNOLI F, GOLINELLI G, et al. Mesenchymal stem/stromal cells as a delivery platform in cell and gene therapies[J]. BMC Med, 2015, 13:186. DOI: 10.1186/s12916-015-0426-0.
    [23] ZHALEH F, AMIRI F, MOHAMMADZADEH-VARDIN M, et al. Nuclear factor erythroid-2 related factor 2 overexpressed mesenchymal stem cells transplantation, improves renal function, decreases injuries markers and increases repair markers in glycerol-induced acute kidney injury rats[J]. Iran J Basic Med Sci, 2016, 19(3): 323-329. https://www.researchgate.net/publication/299452445_Nuclear_factor_erythroid-2_related_factor_2_overexpressed_mesenchymal_stem_cells_transplantation_improves_renal_function_decreases_injuries_markers_and_increases_repair_markers_in_glycerol-induced_Acu
    [24] MOHAMMADZADEH M, HALABIAN R, GHAREHBAGHIAN A, et al. Nrf-2 overexpression in mesenchymal stem cells reduces oxidative stress-induced apoptosis and cytotoxicity[J]. Cell Stress Chaperones, 2012, 17(5): 553-565. DOI: 10.1007/s12192-012-0331-9.
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出版历程
  • 收稿日期:  2017-12-14
  • 网络出版日期:  2021-01-19
  • 刊出日期:  2018-03-15

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