• 1.中國醫(yī)科大學(xué)附屬第一醫(yī)院心臟內(nèi)科（遼寧沈陽 110001）;;
  • 2.遼寧省人民醫(yī)院心臟中心（遼寧沈陽 110016）;

目的探討缺氧誘導(dǎo)因子-1α(hypoxia induced factor-1α,HIF-1α)基因轉(zhuǎn)染是否提高缺氧間充質(zhì)干細胞(mesenchymal stem cells，MSCs)的耐缺氧能力以及這種能力是否與增加葡萄糖攝取有關(guān)。方法將MSCs分為常氧非轉(zhuǎn)染組(即對照組)、常氧轉(zhuǎn)染組、缺氧非轉(zhuǎn)染組和缺氧轉(zhuǎn)染組，分別于常氧(5% CO2，37 ℃)和缺氧(94% N2、1% O2和5% CO2，37 ℃)條件下孵育8 h。免疫細胞化學(xué)、RT-PCR和Western blot法檢測HIF-1α表達,流式細胞儀檢測細胞凋亡率(AR)、死亡率(DR)，MTT法檢測細胞增殖，放射同位素法檢測氚標(biāo)葡萄糖(3H-G)攝取量。結(jié)果①與常氧非轉(zhuǎn)染組相比，常氧轉(zhuǎn)染組HIF-1α mRNA表達明顯增高(P lt;0.01)，但其蛋白表達2組間差異無統(tǒng)計學(xué)意義(P=0.187)； 與缺氧非轉(zhuǎn)染組相比，缺氧轉(zhuǎn)染組的HIF1α mRNA和蛋白表達均明顯增高(P lt;0.01)。②缺氧轉(zhuǎn)染組MSCs的AR(P=0.001)和DR(P=0.003)均明顯低于缺氧非轉(zhuǎn)染組，但均明顯高于常氧非轉(zhuǎn)染組(P lt;0.01)。③與缺氧非轉(zhuǎn)染組相比，缺氧轉(zhuǎn)染組MSCs增殖(P=0.004)明顯提高，但明顯低于常氧非轉(zhuǎn)染組(P=0.001)。④與缺氧非轉(zhuǎn)染組相比，缺氧轉(zhuǎn)染組MSCs對 3H-G的攝取量明顯提高(P=0.004)，但明顯低于常氧非轉(zhuǎn)染組(P=0.001)。⑤AR與HIF-1α蛋白表達(r=－0.71，P=0.005)、3H-G攝取量(r=－0.65，P=0.004)均呈明顯負(fù)相關(guān)，HIF-1α蛋白表達與3H-G攝取量呈明顯正相關(guān)(r=0.77,P=0.003)。結(jié)論HIF-1α基因可顯著提高MSCs的耐缺氧能力，其機理可能與HIF-1α基因提高MSCs對葡萄糖的攝取有關(guān)。

引用本文： 孔宏亮,孫紅波,宋麗杰,齊國先. 缺氧誘導(dǎo)因子-1α過表達對骨髓間充質(zhì)干細胞缺氧時生存能力的影響. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(3): 261-266. doi: 復(fù)制

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2. Wollert KC, Drexler H. Clinical applications of stem cells for the heart [J]. Circ Res, 2005, 96: 151163.
3. Zimmet JM, Hare JM. Emerging role for bone marrow derived mesenchymal stem cells in myocardial regenerative therapy [J]. Basic Res Cardiol, 2005, 100(6): 471481.
4. Minguell JJ, Erices A. Mesenchymal stem cells and the treatment of cardiac disease [J]. Exp Biol Med (Maywood), 2006, 231(1): 3949.
5. 宋來鳳. 從干細胞誘導(dǎo)成心肌細胞研究中的理論和技術(shù)瓶頸 [J]. 中華心血管病雜志, 2005, 33(2): 107108.
6. Kong HL, Liu NN, Huo Xin, et al. Cell multiplication, apoptosis and pAkt protein expression of bone mesenchymal stem cells of rat under hypoxia environment [J]. JNMU, 2007, 21(4): 233239.
7. 孔宏亮, 劉寧寧, 張海鵬, 等. 缺氧環(huán)境下大鼠骨髓間充質(zhì)干細胞的凋亡及Fas、FasL的表達 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2007, 16(5): 538543.
8. 孔宏亮, 劉寧寧, 齊國先, 等. 缺氧對大鼠骨髓間充質(zhì)干細胞增殖、凋亡及葡萄糖攝取和Akt表達的影響 [J]. 中國現(xiàn)代醫(yī)學(xué)雜志, 2007, 18(10): 13741382.
9. 齊國先, 孔宏亮, 劉寧寧, 等. 缺氧環(huán)境下大鼠骨髓間充質(zhì)干細胞凋亡相關(guān)蛋白和mRNA的表達 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2008, 17(2): 138143.
10. 孔宏亮, 劉寧寧, 霍鑫, 等. Akt基因轉(zhuǎn)染對骨髓間充質(zhì)干細胞缺氧時凋亡和增殖的影響 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2008, 17(3): 225231.
11. 霍鑫, 孔宏亮, 胡新華, 等. pEGFPC1/Akt體外轉(zhuǎn)染骨髓間充質(zhì)干細胞后對VEGF表達的影響 [J]. 中華實驗外科雜志, 2007, 12(3): 206211.
12. Kong HL, Li ZQ, Zhao YJ, et al. Cardiac autonomic nerve fiber regeneration in chronic heart failure. Do Akt genetransduced mesenchymal stem cells promote repair? [J]. Neural Regen Res, 2010, 5(1): 2834.
13. Salceda S, Caro J. Hypoxiainducible factor 1α (HIF1α) protein is rapidly degraded by the ubiquitinproteasome system under normoxic conditions. Its stabilization by hypoxia depends on redoxinduced changes [J]. J Biol Chem, 1997, 272(36): 2264222647.
14. Soucek T, Cumming R, Dargusch R, et al. The regulation of glucose metabolism by HIF1 mediates a neuroprotective response to amyloid beta peptide [J]. Neuron, 2003, 39(1): 4356.
15. 胡鳶, 楊柳, 段小軍. 缺氧誘導(dǎo)因子1α基因轉(zhuǎn)染人間充質(zhì)干細胞 [J]. 第三軍醫(yī)大學(xué)學(xué)報, 2005, 27(9): 928929.
16. 胡鳶, 段小軍, 楊柳. 缺氧誘導(dǎo)因子1α基因轉(zhuǎn)染人間充質(zhì)干細胞的體內(nèi)促血管生成作用 [J]. 中華實驗外科雜志, 2006, 23(8): 993996.
17. Zheng X, Ruas JL, Cao R, et al. Celltypespecific regulation of degradation of hypoxiainducible factor 1α: role of subcellular compartmentalization [J]. Mol Cell Biol, 2006, 26(12): 46284641.
18. 邵磊, 周書春, 徐桂萍, 等. 體外Akt基因轉(zhuǎn)染對骨髓間充質(zhì)干細胞葡萄糖轉(zhuǎn)運載體4表達和易位的影響 [J]. 中國普外基礎(chǔ)與臨床雜志, 2010, 17(1): 5256.
19. Schipani E. Posttranslational modifications of collagens as targets of hypoxia and Hif1α in endochondral bone development [J]. Ann N Y Acad Sci, 2010, 1192: 317321.
20. Westra J, Molema G, Kallenberg CG. Hypoxiainducible factor1 as regulator of angiogenesis in rheumatoid arthritis-therapeutic implications [J]. Curr Med Chem, 2010, 17(3): 254263.
21. Rey S, Semenza GL. Hypoxiainducible factor1dependent mechanisms of vascularization and vascular remodelling [J]. Cardiovasc Res, 2010, 86(2): 236242.
22. Heddleston JM, Li Z, Lathia JD, et al. Hypoxia inducible factors in cancer stem cells [J]. Br J Cancer, 2010, 102(5): 789795.
23. Lin J, Denmeade S, Carducci MA. HIF1α and calcium signaling as targets for treatment of prostate cancer by cardiac glycosides [J]. Curr Cancer Drug Targets, 2009, 9(7): 881887.
  1. 1. Aceves JL, Archundia A, Díaz G, et al. Stem cell perspectives in myocardial infarctions [J]. Rev Invest Clin, 2005, 57(2): 156162.
  2. 2. Wollert KC, Drexler H. Clinical applications of stem cells for the heart [J]. Circ Res, 2005, 96: 151163.
  3. 3. Zimmet JM, Hare JM. Emerging role for bone marrow derived mesenchymal stem cells in myocardial regenerative therapy [J]. Basic Res Cardiol, 2005, 100(6): 471481.
  4. 4. Minguell JJ, Erices A. Mesenchymal stem cells and the treatment of cardiac disease [J]. Exp Biol Med (Maywood), 2006, 231(1): 3949.
  5. 5. 宋來鳳. 從干細胞誘導(dǎo)成心肌細胞研究中的理論和技術(shù)瓶頸 [J]. 中華心血管病雜志, 2005, 33(2): 107108.
  6. 6. Kong HL, Liu NN, Huo Xin, et al. Cell multiplication, apoptosis and pAkt protein expression of bone mesenchymal stem cells of rat under hypoxia environment [J]. JNMU, 2007, 21(4): 233239.
  7. 7. 孔宏亮, 劉寧寧, 張海鵬, 等. 缺氧環(huán)境下大鼠骨髓間充質(zhì)干細胞的凋亡及Fas、FasL的表達 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2007, 16(5): 538543.
  8. 8. 孔宏亮, 劉寧寧, 齊國先, 等. 缺氧對大鼠骨髓間充質(zhì)干細胞增殖、凋亡及葡萄糖攝取和Akt表達的影響 [J]. 中國現(xiàn)代醫(yī)學(xué)雜志, 2007, 18(10): 13741382.
  9. 9. 齊國先, 孔宏亮, 劉寧寧, 等. 缺氧環(huán)境下大鼠骨髓間充質(zhì)干細胞凋亡相關(guān)蛋白和mRNA的表達 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2008, 17(2): 138143.
  10. 10. 孔宏亮, 劉寧寧, 霍鑫, 等. Akt基因轉(zhuǎn)染對骨髓間充質(zhì)干細胞缺氧時凋亡和增殖的影響 [J]. 中國組織化學(xué)與細胞化學(xué)雜志, 2008, 17(3): 225231.
  11. 11. 霍鑫, 孔宏亮, 胡新華, 等. pEGFPC1/Akt體外轉(zhuǎn)染骨髓間充質(zhì)干細胞后對VEGF表達的影響 [J]. 中華實驗外科雜志, 2007, 12(3): 206211.
  12. 12. Kong HL, Li ZQ, Zhao YJ, et al. Cardiac autonomic nerve fiber regeneration in chronic heart failure. Do Akt genetransduced mesenchymal stem cells promote repair? [J]. Neural Regen Res, 2010, 5(1): 2834.
  13. 13. Salceda S, Caro J. Hypoxiainducible factor 1α (HIF1α) protein is rapidly degraded by the ubiquitinproteasome system under normoxic conditions. Its stabilization by hypoxia depends on redoxinduced changes [J]. J Biol Chem, 1997, 272(36): 2264222647.
  14. 14. Soucek T, Cumming R, Dargusch R, et al. The regulation of glucose metabolism by HIF1 mediates a neuroprotective response to amyloid beta peptide [J]. Neuron, 2003, 39(1): 4356.
  15. 15. 胡鳶, 楊柳, 段小軍. 缺氧誘導(dǎo)因子1α基因轉(zhuǎn)染人間充質(zhì)干細胞 [J]. 第三軍醫(yī)大學(xué)學(xué)報, 2005, 27(9): 928929.
  16. 16. 胡鳶, 段小軍, 楊柳. 缺氧誘導(dǎo)因子1α基因轉(zhuǎn)染人間充質(zhì)干細胞的體內(nèi)促血管生成作用 [J]. 中華實驗外科雜志, 2006, 23(8): 993996.
  17. 17. Zheng X, Ruas JL, Cao R, et al. Celltypespecific regulation of degradation of hypoxiainducible factor 1α: role of subcellular compartmentalization [J]. Mol Cell Biol, 2006, 26(12): 46284641.
  18. 18. 邵磊, 周書春, 徐桂萍, 等. 體外Akt基因轉(zhuǎn)染對骨髓間充質(zhì)干細胞葡萄糖轉(zhuǎn)運載體4表達和易位的影響 [J]. 中國普外基礎(chǔ)與臨床雜志, 2010, 17(1): 5256.
  19. 19. Schipani E. Posttranslational modifications of collagens as targets of hypoxia and Hif1α in endochondral bone development [J]. Ann N Y Acad Sci, 2010, 1192: 317321.
  20. 20. Westra J, Molema G, Kallenberg CG. Hypoxiainducible factor1 as regulator of angiogenesis in rheumatoid arthritis-therapeutic implications [J]. Curr Med Chem, 2010, 17(3): 254263.
  21. 21. Rey S, Semenza GL. Hypoxiainducible factor1dependent mechanisms of vascularization and vascular remodelling [J]. Cardiovasc Res, 2010, 86(2): 236242.
  22. 22. Heddleston JM, Li Z, Lathia JD, et al. Hypoxia inducible factors in cancer stem cells [J]. Br J Cancer, 2010, 102(5): 789795.
  23. 23. Lin J, Denmeade S, Carducci MA. HIF1α and calcium signaling as targets for treatment of prostate cancer by cardiac glycosides [J]. Curr Cancer Drug Targets, 2009, 9(7): 881887.