欢迎访问《解剖学报》官方网站！今天是
English
趋化因子-8对高糖环境下脂肪间充质干细胞迁移能力的影响
Effect of chemokine-8 factor on the migration ability of human adipose derived mesenchymal stem cell in high glucose environment
目的 探讨高糖环境下，趋化因子-8（CXCL-8）对人脂肪间充质干细胞（hADMSCs）迁移能力的影响和机制。方法 建立高糖环境模型；在高糖条件下，建立CXCL-8实验组、Akt抑制剂组和高糖对照组，在正常条件培养的hADMSCs为正常对照组；分别用细胞划痕实验、Transwell细胞小室实验检测CXCL-8对hADMSCs的迁移能力影响，并用Western blotting、ELISA实验检测Akt、信号转导和转录激活因子（STAT3）和血管内皮生长因子（VEGF）等蛋白表达。
结果 相对高糖对照组，CXCL-8实验组hADMSCs细胞划痕面积闭合率和Transwell细胞小室迁移率均增高（P＜0.01）；而Akt抑制剂组hADMSCs细胞划痕面积闭合率或Transwell细胞小室迁移率比CXCL-8实验组皆降低（P＜0.01）；CXCL-8实验组磷酸化Akt、mTOP、STAT3蛋白表达增高，CXCL-8实验组上清液的VEGF、表皮细胞生长因子 (EGF)含量明显增高（P＜0.01）；而Akt抑制剂组hADMSCs分泌能力下降（P＜0.01）。 结论 高糖环境下，CXCL-8通过Akt-STAT3通路促进间充质干细胞（MSCs)旁分泌VEGF等因子，促进MSCs迁移，对招募宿主细胞归巢，促进组织损伤修复具有重要意义。
Objective To investigate the effect of chemokines (CXCL)-8 factor on the chemotaxis ability of human adipose-derived mesenchymal stem cells (hADMSCs).
Methods The animals under the high glucose environment were divided into CXCL-8 experimental group, Akt inhibitor group and control group of high glucose. The normal control group was the cultural hADMSCs under the normal condition. Cell scratch, Transwells cell chamber experiment were used to check the effect of chemotaxis ability of CXCL-8 on the ADMSCs Western blotting, and ELISA experiment to check the protein expression of Akt, signal transducer and activator of transcription 3 (STAT3) and vascular endothelial growth factor (VEGF). Results Compared with the control group, the closing rate of hADMSCs cell scratch area in CXCL-8 experimental group and migration rate of Transwell cell chamber were increased (P＜0.01). The closing rate of cell scratch area in Akt inhibitor group and migration rate were decreased (P＜0.01). The protein expressions of phosphorylated Akt, mTOP and STAT3 were increased. The VEGF, epidermal growth factor (EGF) of liquid supernatant in CXCL-8 experimental group were increased significantly (P＜0.01); but the secretion ability of Akt inhibitor group was decreased (P＜0.01). Conclusion Under the high glucose environment, the CXCL-8 promoted MSCs paracrine VEGF factors through Akt-STAT3 pathway and promoted migration of MSCs, which may have the important meaning for recruit host cell homing and promote damage repair.
趋化因子-8 / 脂肪间充质干细胞 / 高血糖环境 / 旁分泌 / 归巢 / 人
Chemokines-8 / Adipose-derived mesenchymal stem cell / High glucose environment / Paracrine / Homing / Human
［1］Guariguata L, Whiting DR, Hambleton I, et al. Global estimates of diabetes prevalence for 2013 and projections for 2035 ［J］. Diabetes Res Clin Pract, 2014, 103(2): 137-149.［2］Gibbons CH, Freeman R. Treatment-induced neuropathy of diabetes: an acute, iatrogenic complication of diabetes ［J］. Brain, 2015, 138(1): 43-52.
［3］Yang W, Lu J, Weng J, et al. Prevalence of diabetes among men and women in China ［J］. N Engl J Med, 2010, 362(12): 1090-1101.
［4］Hopkins RB, Burke N, Harlock J, et al. Economic burden of illness associated with diabetic foot ulcers in Canada ［J］. BMC Health Serv Res, 2015, 15(1): 13.
［5］Tae SK, Lee SH, Park JS, et al. Mesenchymal stem cells for tissue engineering and regenerative medicine ［J］. Biomed Mater, 2006, 1(2): 63-71.
［6］Wittenburg G, Flade V, Garbe AI, et al. Scaffold preferences of mesenchymal stromal cells and adipose-derived stem cells from green fluorescent protein transgenic mice influence the tissue engineering of bone［J］. Br J Oral Maxillofac Surg, 2014, 52(5): 409-414.
［7］Sargent J. Diabetes: Functional impairment of bone marrow progenitor cells in diabetes mellitus ［J］. Nat Rev Endocrinol, 2014, 10(7): 379.
［8］Bischoff DS, Zhu JH, Makhijani NS, et al. Acidic pH stimulates the production of the angiogenic CXC chemokine, CXCL8 (interleukin-8), in human adult mesenchymal stem cells via the extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and NF-kappaB pathways ［J］. J Cell Biochem, 2008, 104(4): 1378-1392.
［9］Russo RC, Garcia CC, Teixeira MM, et al. The CXCL8/IL-8 chemokine family and its receptors in inflammatory diseases ［J］. Expert Rev Clin Immunol, 2014, 10(5): 593-619.
［10］Piperi C, Samaras V, Levidou G, et al. Prognostic significance of IL-8-STAT-3 pathway in astrocytomas: correlation with IL-6, VEGF and microvessel morphometry ［J］. Cytokine, 2011, 55(3): 387-395.
［11］Barcelos LS, Duplaa C, Krankel N, et al. Human CD133+ progenitor cells promote the healing of diabetic ischemic ulcers by paracrine stimulation of angiogenesis and activation of Wnt signaling ［J］. Circ Res, 2009, 104(9): 1095-1102.
［12］Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro ［J］. Nat Protoc, 2007, 2(2): 329-333.
［13］Marshall J. Transwell (R) invasion assays ［J］. Methods Mol Biol, 2011, 769: 97-110.
［14］Wang YL, He XM, Tang W, et al. Stromal cell-derived factor-1-α / CXCR4/ CXCR7 axis of the bone marrow mesenchymal stem cell migration of affect ［J］. Acta Anatomica Sinica, 2014, 45(5): 639-645. (in Chinese)
王玉兰, 何晓梅, 唐薇, 等. 基质细胞衍生因子-1-α/CXCR4/CXCR7轴对骨髓间充质干细胞迁移的影响 ［J］. 解剖学报, 2014, 45(5): 639-645.
［15］Li D, Wang N, Zhang L, et al. Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor ［J］. Stem Cell Res Ther, 2013, 4(5): 103.
［16］Kim H, Choi K, Kweon OK, et al. Enhanced wound healing effect of canine adipose-derived mesenchymal stem cells with low-level laser therapy in athymic mice ［J］. J Dermatol Sci, 2012, 68(3): 149-156.
［17］Shestopalov IA, Zon LI. Stem cells: The right neighbour ［J］. Nature, 2012, 481(7382): 453-455.
［18］Dona E, Barry JD, Valentin G, et al. Directional tissue migration through a self-generated chemokine gradient ［J］. Nature, 2013, 503(7475): 285-289.
［19］Liu LB, Xie F, Chang KK, et al. Hypoxia promotes the proliferation of cervical carcinoma cells through stimulating the secretion of IL8 ［J］. Int J Clin Exp Pathol, 2014, 7(2): 575-583.
［20］Sun BK, Siprashvili Z, Khavari PA. Advances in skin grafting and treatment of cutaneous wounds ［J］. Science, 2014, 346(6212): 941-945.
［21］Sawangmake C, Pavasant P, Chansiripornchai P, et al. High glucose condition suppresses neurosphere formation by human periodontal ligament-derived mesenchymal stem cells ［J］. J Cell Biochem, 2014, 115(5): 928-939.
［22］Hou Y, Ryu CH, Jun JA, et al. IL-8 enhances the angiogenic potential of human bone marrow mesenchymal stem cells by increasing vascular endothelial growth factor ［J］. Cell Biol Int, 2014, 38(9): 1050-1059.
［23］Bi LK, Zhou N, Liu C, et al. Kidney cancer cells secrete IL-8 to activate Akt and promote migration of mesenchymal stem cells ［J］. Urol Oncol, 2014, 32(5): 607-612.
［24］Tomanek RJ, Christensen LP, Simons M, et al. Embryonic coronary vasculogenesis and angiogenesis are regulated by interactions between multiple FGFs and VEGF and are influenced by mesenchymal stem cells ［J］. Dev Dyn, 2010, 239(12): 3182-3191.
［25］Kozikowski AP, Sun H, Brognard J, et al. Novel PI analogues selectively block activation of the pro-survival serine/threonine kinase Akt ［J］. J Am Chem Soc, 2003, 125(5): 1144-1145.
［26］Hou C, Shen L, Huang Q, et al. The effect of heme oxygenase-1 complexed with collagen on MSC performance in the treatment of diabetic ischemic ulcer ［J］. Biomaterials, 2013, 34(1): 112-120.
［27］Shen L, Zeng W, Wu YX, et al. Neurotrophin-3 accelerates wound healing in diabetic mice by promoting a paracrine response in mesenchymal stem cells ［J］. Cell Transplant, 2013, 22(6): 1011-1021.
［28］Tang J, Wang J, Zheng F, et al. Combination of chemokine and angiogenic factor genes and mesenchymal stem cells could enhance angiogenesis and improve cardiac function after acute myocardial infarction in rats ［J］. Mol Cell Biochem, 2010, 339(12): 107-118.
［29］Chao YH, Wu KH, Chiou SH, et al. Downregulated CXCL12 expression in mesenchymal stem cells associated with severe aplastic anemia in children ［J］. Ann Hematol, 2015, 94(1): 13-22.
黑龙江省教育厅科学技术研究项目面上课题
/
| 〈 |
|
〉 |
