Role and mechanism of stromal cell derived factor 1 on proliferation of vascular endothelial cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LIU Zongchao ¹ , WEI Zhangchao ¹ , LIU Yong ¹ , WANG Zhenlong ¹ , FU Zhijiang ¹ , LIU Shigui ¹ , SHEN Jieliang ² ,  HU Zhenming ²

1. Department of Orthopaedics, the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou Sichuan, 646000, P.R.China;
2. Department of Orthopaedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400042, P.R.China;

Corresponding author：

HU Zhenming, Email: spinecenter_hu@126.com

Keywords：

Stromall cell derived factor 1; nucleus pulposus cells; vascular endothelial cells; three-dimensional culture; Akt

DOI：

10.7507/1002-1892.201609071

Video：

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Objective To investigate the role and relative mechanism of stromal cell derived factorl (SDF-1) secreted by nucleus pulposus cells (NPCs) on the proliferation of vascular endothelial cells (VECs). Methods The NPCs were isolated from the degenerated disc specimens after discectomy. NPCs at passage 1 were transfected with lentivirus-mediated SDF-1 over-expression; transfected and untransfected NPCs at passage 2 were cultured in the three-dimensional alvetex® scaffold, then they were co-cultured with HMEC-1 cells. The morphology of NPCs was observed by scanning electron microscope (SEM), and the apoptosis of HMEC-1 cells was detected by Annexin V/propidiumiodide staining after 72 hours co-culutre. The proliferation of HMEC-1 cells was detected by cell counting kit 8 at 12, 24, 48, and 72 hours in transfected group and untransfected group, respectively. ELISA was used to measure the vascular endothelial growth factor (VEGF) expression level. The virus transfection efficiency and relative Akt pathway were determined by Western blot. Results The NPCs maintained cell phenotype and secreted much extracellular matrix in three-dimensional-culture by SEM observation. In the co-culutre system, after NPCs were transfected with SDF-1 over-expression lentivirus, the proliferation of HMEC-1 cells was significantly increased, while the apoptosis was decreased obviously. The ELISA results demonstrated that the amount of VEGF was remarkably increased in the culture medium. Furthermore, SDF-1 promoted the up-regulation of phosphorylate Akt expression; after inhibition of Akt expression by GSK690693, the proliferation rate of VECs decreased significantly. Conclusion Over-expression of SDF-1 by NPCs is beneficial for VECs proliferation, which is involved in SDF-1-Akt signalling pathway.

Citation： LIUZongchao, WEIZhangchao, LIUYong, WANGZhenlong, FUZhijiang, LIUShigui, SHENJieliang, HUZhenming. Role and mechanism of stromal cell derived factor 1 on proliferation of vascular endothelial cells. Chinese Journal of Reparative and Reconstructive Surgery, 2017, 31(1): 91-97. doi: 10.7507/1002-1892.201609071 Copy

1.	Karamouzian S, Eskandary H, Faramarzee M, et al. Frequency of lumbar intervertebral disc calcification and angiogenesis, and their correlation with clinical, surgical, and magnetic resonance imaging findings. Spine (Phila Pa 1976), 2010, 35(8): 881-886.
2.	Ohba T, Haro H, Ando T, et al. TNF-alpha-induced NF-kappaB signaling reverses age-related declines in VEGF induction and angiogenic activity in intervertebral disc tissues. J Orthop Res, 2009, 27(2): 229-235.
3.	Vernon-Roberts B, Moore RJ, Fraser RD. The natural history of age-related disc degeneration: the pathology and sequelae of tears. Spine (Phila Pa 1976), 2007, 32(25): 2797-2804.
4.	Pohl PH, Lozito TP, Cuperman T, et al. Catabolic effects of endothelial cell-derived microparticles on disc cells: Implications in intervertebral disc neovascularization and degeneration. J Orthop Res, 2016, 34(8): 1466-1474.
5.	Kim JH, Studer RK, Vo NV, et al. p38 MAPK inhibition selectively mitigates inflammatory mediators and VEGF production in AF cells co-cultured with activated macrophage-like THP-1 cells. Osteoarthritis Cartilage, 2009, 17(12): 1662-1669.
6.	Lee JM, Song JY, Baek M, et al. Interleukin-1beta induces angiogenesis and innervation in human intervertebral disc degeneration. J Orthop Res, 2011, 29(2): 265-269.
7.	Liu Z, Ma C, Shen J, et al. SDF1/CXCR4 axis induces apoptosis of human degenerative nucleus pulposus cells via the NFkappaB pathway. Mol Med Rep, 2016, 14(1): 783-789.
8.	Hiasa K, Ishibashi M, Ohtani K, et al. Gene transfer of stromal cell-derived factor-1alpha enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/endothelial nitric oxide synthase-related pathway: next-generation chemokine therapy for therapeutic neovascularization. Circulation, 2004, 109(20): 2454-2461.
9.	Pereira CL, Goncalves RM, Peroglio M, et al. The effect of hyaluronan-based delivery of stromal cell-derived factor-1 on the recruitment of MSCs in degenerating intervertebral discs. Biomaterials, 2014, 35(28): 8144-8153.
10.	Zhang H, Zhang L, Chen L, et al. Stromal cell-derived factor-1 and its receptor CXCR4 are upregulated expression in degenerated intervertebral discs. Int J Med Sci, 2014, 11(3): 240-245.
11.	Simon J, McAuliffe M, Shamim F, et al. Discogenic low back pain. Phys Med Rehabil Clin N Am, 2014, 25(2): 305-317.
12.	Yu N, Zhang Z, Chen P, et al. Tetramethylpyrazine (TMP), an Active Ingredient of Chinese Herb Medicine Chuanxiong, Attenuates the Degeneration of Trabecular Meshwork through SDF-1/CXCR4 Axis. PLoS One, 2015, 10(8): e0133055.
13.	Ascione R, Rowlinson J, Avolio E, et al. Migration towards SDF-1 selects angiogenin-expressing bone marrow monocytes endowed with cardiac reparative activity in patients with previous myocardial infarction. Stem Cell Res Ther, 2015, 6: 53.
14.	Jia CQ, Zhao JG, Zhang SF, et al. Stromal cell-derived factor-1 and vascular endothelial growth factor may play an important role in the process of neovascularization of herniated intervertebral discs. J Int Med Res, 2009, 37(1): 136-144.
15.	Chang TT, Wu TC, Huang PH, et al. Direct Renin Inhibition with Aliskiren Improves Ischemia-Induced Neovasculogenesis in Diabetic Animals via the SDF-1 Related Mechanism. PLoS One, 2015, 10(8): e0136627.

1. Karamouzian S, Eskandary H, Faramarzee M, et al. Frequency of lumbar intervertebral disc calcification and angiogenesis, and their correlation with clinical, surgical, and magnetic resonance imaging findings. Spine (Phila Pa 1976), 2010, 35(8): 881-886.
2. Ohba T, Haro H, Ando T, et al. TNF-alpha-induced NF-kappaB signaling reverses age-related declines in VEGF induction and angiogenic activity in intervertebral disc tissues. J Orthop Res, 2009, 27(2): 229-235.
3. Vernon-Roberts B, Moore RJ, Fraser RD. The natural history of age-related disc degeneration: the pathology and sequelae of tears. Spine (Phila Pa 1976), 2007, 32(25): 2797-2804.
4. Pohl PH, Lozito TP, Cuperman T, et al. Catabolic effects of endothelial cell-derived microparticles on disc cells: Implications in intervertebral disc neovascularization and degeneration. J Orthop Res, 2016, 34(8): 1466-1474.
5. Kim JH, Studer RK, Vo NV, et al. p38 MAPK inhibition selectively mitigates inflammatory mediators and VEGF production in AF cells co-cultured with activated macrophage-like THP-1 cells. Osteoarthritis Cartilage, 2009, 17(12): 1662-1669.
6. Lee JM, Song JY, Baek M, et al. Interleukin-1beta induces angiogenesis and innervation in human intervertebral disc degeneration. J Orthop Res, 2011, 29(2): 265-269.
7. Liu Z, Ma C, Shen J, et al. SDF1/CXCR4 axis induces apoptosis of human degenerative nucleus pulposus cells via the NFkappaB pathway. Mol Med Rep, 2016, 14(1): 783-789.
8. Hiasa K, Ishibashi M, Ohtani K, et al. Gene transfer of stromal cell-derived factor-1alpha enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/endothelial nitric oxide synthase-related pathway: next-generation chemokine therapy for therapeutic neovascularization. Circulation, 2004, 109(20): 2454-2461.
9. Pereira CL, Goncalves RM, Peroglio M, et al. The effect of hyaluronan-based delivery of stromal cell-derived factor-1 on the recruitment of MSCs in degenerating intervertebral discs. Biomaterials, 2014, 35(28): 8144-8153.
10. Zhang H, Zhang L, Chen L, et al. Stromal cell-derived factor-1 and its receptor CXCR4 are upregulated expression in degenerated intervertebral discs. Int J Med Sci, 2014, 11(3): 240-245.
11. Simon J, McAuliffe M, Shamim F, et al. Discogenic low back pain. Phys Med Rehabil Clin N Am, 2014, 25(2): 305-317.
12. Yu N, Zhang Z, Chen P, et al. Tetramethylpyrazine (TMP), an Active Ingredient of Chinese Herb Medicine Chuanxiong, Attenuates the Degeneration of Trabecular Meshwork through SDF-1/CXCR4 Axis. PLoS One, 2015, 10(8): e0133055.
13. Ascione R, Rowlinson J, Avolio E, et al. Migration towards SDF-1 selects angiogenin-expressing bone marrow monocytes endowed with cardiac reparative activity in patients with previous myocardial infarction. Stem Cell Res Ther, 2015, 6: 53.
14. Jia CQ, Zhao JG, Zhang SF, et al. Stromal cell-derived factor-1 and vascular endothelial growth factor may play an important role in the process of neovascularization of herniated intervertebral discs. J Int Med Res, 2009, 37(1): 136-144.
15. Chang TT, Wu TC, Huang PH, et al. Direct Renin Inhibition with Aliskiren Improves Ischemia-Induced Neovasculogenesis in Diabetic Animals via the SDF-1 Related Mechanism. PLoS One, 2015, 10(8): e0136627.

Chinese Journal of Reparative and Reconstructive Surgery

Role and mechanism of stromal cell derived factor 1 on proliferation of vascular endothelial cells

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