Effect of Hypoxia Induced Factor-1α Over-Expression on Survival Capacity of Hypoxia Bone Marrow Mesenchymal Stem Cells Ex Vivo_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

KONG Hongliang ^1,2 , SUN Hongbo ² , SONG Lijie ² , QI Guoxian ¹

1.Department of Cardiology, The First Affiliated Hospital, China Medical University, Shenyang 110001, Liaoning Province, China;;
2.Center of Cardiology, The People’s Hospital of Liaoning Province, Shenyang 110016, Liaoning Province, China;

Corresponding author：

Keywords：

Bone marrow mesenchymal stem cell; Hypoxia induced factor-1α gene; Transfection; Hypoxia; Apoptosis; Glucose uptake; Rat

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ObjectiveTo elucidate whether hypoxia induced factor-1α (HIF-1α) gene improved hypoxia tolerant capability of bone marrow mesenchymal stem cells uptake(MSCs) or not and whether the capability was related to glucose uptake increase in hypoxia MSCs ex vivo or not. MethodsMSCs were randomly divided into normoxia non-HIF-1α transfection group (control group), normoxia HIF-1α transfection group, hypoxia non-HIF-1α transfection group, and hypoxia HIF-1α transfection group and then each group was cultured with normoxia (5% CO2 at 37 ℃) or hypoxia (94% N2, 1% O2, 5% CO2 at 37 ℃) for 8 h, respectively. Finally, the expressions of HIF-1α were detected by immunocytochemistry, RT-PCR, and Western blot methods, respectively. Apoptosis ratio (AR) and death ratio (DR) were tested by flow cytometry. The proliferation was detected by MTT method. Glucose uptake was assayed by radiation isotope method. Results① Compared with the normoxia non-HIF-1α transfection group, the expression of HIF-1α mRNA significantly increased (P lt;0.01) in the normoxia HIF-1α transfection group except for its protein (P=0.187); Both of mRNA and protein expressions of HIF-1α in the hypoxia HIF-1α transfection group were significantly higher than those in the hypoxia non-HIF-1α transfection group (P lt;0.01). ② The AR (P=0.001) and DR (P=0.003) in the hypoxia HIF-1α transfection group were significantly lower than
those in the hypoxia non-HIF-1α transfection group, both of which were significantly higher than those in the normoxia non-HIF-1α transfection group (P lt;0.01). ③ The proliferation of MSCs in the hypoxia HIF-1α transfection group was significantly higher than that in the hypoxia non-HIF-1α transfection group (P=0.004), which significantly lower than that in the normoxia non-HIF-1α transfection group (P=0.001). ④ Compared with the hypoxia non-HIF-1α transfection group, the 3H-G uptake capability (P=0.004) of MSCs significantly increased in the hypoxia HIF-1α transfection group, which was significantly lower than that in the normoxia non-HIF-1α transfection group (P=0.001). ⑤ There were significantly negative relation between AR and HIF-1α protein (r=－0.71，P=0.005) or 3H-G uptake (r=－0.65，P=0.004), and significantly positive relation between HIF-1α protein expression and 3H-G uptake (r=0.77, P=0.003). ConclusionHIF-1α gene significantly improves anti-hypoxia capability of MSCs, which is fulfilled by increasing glucose upake.

Citation： KONG Hongliang ,SUN Hongbo,SONG Lijie,QI Guoxian. Effect of Hypoxia Induced Factor-1α Over-Expression on Survival Capacity of Hypoxia Bone Marrow Mesenchymal Stem Cells Ex Vivo. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2011, 18(3): 261-266. doi: Copy

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.	Wollert KC, Drexler H. Clinical applications of stem cells for the heart [J]. Circ Res, 2005, 96: 151163.
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7.	孔宏亮, 刘宁宁, 张海鹏, 等. 缺氧环境下大鼠骨髓间充质干细胞的凋亡及Fas、FasL的表达 [J]. 中国组织化学与细胞化学杂志, 2007, 16(5): 538543.
8.	孔宏亮, 刘宁宁, 齐国先, 等. 缺氧对大鼠骨髓间充质干细胞增殖、凋亡及葡萄糖摄取和Akt表达的影响 [J]. 中国现代医学杂志, 2007, 18(10): 13741382.
9.	齐国先, 孔宏亮, 刘宁宁, 等. 缺氧环境下大鼠骨髓间充质干细胞凋亡相关蛋白和mRNA的表达 [J]. 中国组织化学与细胞化学杂志, 2008, 17(2): 138143.
10.	孔宏亮, 刘宁宁, 霍鑫, 等. Akt基因转染对骨髓间充质干细胞缺氧时凋亡和增殖的影响 [J]. 中国组织化学与细胞化学杂志, 2008, 17(3): 225231.
11.	霍鑫, 孔宏亮, 胡新华, 等. pEGFPC1/Akt体外转染骨髓间充质干细胞后对VEGF表达的影响 [J]. 中华实验外科杂志, 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.	胡鸢, 杨柳, 段小军. 缺氧诱导因子1α基因转染人间充质干细胞 [J]. 第三军医大学学报, 2005, 27(9): 928929.
16.	胡鸢, 段小军, 杨柳. 缺氧诱导因子1α基因转染人间充质干细胞的体内促血管生成作用 [J]. 中华实验外科杂志, 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, 周书春, 徐桂萍, 等. 体外Akt基因转染对骨髓间充质干细胞葡萄糖转运载体4表达和易位的影响 [J]. 中国普外基础与临床杂志, 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. Aceves JL, Archundia A, Díaz G, et al. Stem cell perspectives in myocardial infarctions [J]. Rev Invest Clin, 2005, 57(2): 156162.
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. 宋来凤. 从干细胞诱导成心肌细胞研究中的理论和技术瓶颈 [J]. 中华心血管病杂志, 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, 刘宁宁, 张海鹏, 等. 缺氧环境下大鼠骨髓间充质干细胞的凋亡及Fas、FasL的表达 [J]. 中国组织化学与细胞化学杂志, 2007, 16(5): 538543.
8. 孔宏亮, 刘宁宁, 齐国先, 等. 缺氧对大鼠骨髓间充质干细胞增殖、凋亡及葡萄糖摄取和Akt表达的影响 [J]. 中国现代医学杂志, 2007, 18(10): 13741382.
9. 齐国先, 孔宏亮, 刘宁宁, 等. 缺氧环境下大鼠骨髓间充质干细胞凋亡相关蛋白和mRNA的表达 [J]. 中国组织化学与细胞化学杂志, 2008, 17(2): 138143.
10. 孔宏亮, 刘宁宁, 霍鑫, 等. Akt基因转染对骨髓间充质干细胞缺氧时凋亡和增殖的影响 [J]. 中国组织化学与细胞化学杂志, 2008, 17(3): 225231.
11. 霍鑫, 孔宏亮, 胡新华, 等. pEGFPC1/Akt体外转染骨髓间充质干细胞后对VEGF表达的影响 [J]. 中华实验外科杂志, 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. 胡鸢, 杨柳, 段小军. 缺氧诱导因子1α基因转染人间充质干细胞 [J]. 第三军医大学学报, 2005, 27(9): 928929.
16. 胡鸢, 段小军, 杨柳. 缺氧诱导因子1α基因转染人间充质干细胞的体内促血管生成作用 [J]. 中华实验外科杂志, 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, 周书春, 徐桂萍, 等. 体外Akt基因转染对骨髓间充质干细胞葡萄糖转运载体4表达和易位的影响 [J]. 中国普外基础与临床杂志, 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.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of Hypoxia Induced Factor-1α Over-Expression on Survival Capacity of Hypoxia Bone Marrow Mesenchymal Stem Cells Ex Vivo

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