孙少松;郑欣;姚圣城;褚耿磊;庞勇;董红燕;赵凤朝;郭开今;

• 1:徐州医科大学附属医院骨科
• 2:徐州医科大学研究生学院
• 3:徐州医科大学神经生物学实验室

摘要(Abstract)：

[目的]研究色素上皮衍生因子(pigment epithelium-derived factor,PEDF)在高浓度地塞米松刺激下,体外培养的成骨细胞迁移及分化的影响,探讨PEDF对成骨细胞功能的保护作用。[方法]将体外培养的MC3T3-E1细胞分为六组,分别为对照组、高浓度地塞米松(dexamethasone,DEX 10-6mol/L)组,PEDF干预地塞米松组,PEDF干预地塞米松组又按照PEDF的浓度分为3组(2、10、50 nmol/L)及成骨诱导培养组。采用Transwell小室及划痕实验于48 h后观察细胞的迁移情况。于14 d时采用倒置相差显微镜观察细胞形态的变化,并进行细胞化学染色(茜素红染色、油红O染色),运用Western Blot检测细胞内PPARγ-2、Runx2蛋白的表达。[结果]在高浓度地塞米松作用下,细胞迁移能力受到抑制,细胞分化能力减弱,胞浆中出现脂滴,Western Blot结果显示脂肪细胞标志蛋白PPARγ-2表达增高,成骨细胞标志蛋白Runx2表达减少。而PEDF干预后可明显逆转高浓度地塞米松抑制细胞迁移及分化的作用。[结论]大剂量应用糖皮质激素可能抑制成骨细胞的迁移及成骨分化能力,促进成骨细胞转分化为脂肪细胞,而PEDF可以有效地逆转这一过程,起到保护细胞功能的作用。

关键词(KeyWords)： 地塞米松;色素上皮衍生因子;成骨细胞;迁移;分化

Abstract:

Keywords:

基金项目(Foundation): 江苏省卫生计生委面上科研课题(编号:H201528);; 江苏省级重点研发专项资金项目(编号:BE2015627);; 江苏省卫生厅科技项目(编号:Q201201)

作者(Author): 孙少松;郑欣;姚圣城;褚耿磊;庞勇;董红燕;赵凤朝;郭开今;

Email:

DOI:

参考文献(References)：

• [1]Migliaccio S,Brama M,Malavolta N.Management of glucocorticoids-induced osteoporosis:role of teriparatide[J].Ther Clin Risk Manag,2009,5(2):305-310.
• [2]Kenanidis E,Potoupnism E,Kakoulidi SP,et al.Management of glucocorticoid-induced osteoporosis:clinical data in relation to disease demographics,bone mineral density and fracture risk[J].Expert Opin Drug Saf,2015,14(7):1035-1053.
• [3]He X,Cheng R,Benyajati S,et al.PEDF and its roles in physiological and pathological conditions:implication in diabetic and hypoxiainduced angiogenic diseases[J].Clin Sci(Lond),2015,128(11):805-823.
• [4]Broadhead ML,Akiyama T,Choong PF,et al.The pathophysiological role of PEDF in bone diseases[J].Curr Mol Med,2010,10(3):296-301.
• [5]Lee YK,Song J,Lee SB,et al.Proliferation,differentiation,and calcification of preosteoblast-like MC3T3-E1 cells cultured onto noncrystalline calcium phosphate glass[J].J Biomed Mater Res A,2004,69(1):188-195.
• [6]Craword S E,Fitchev P,Veliceasa D,et al.The many facets of PEDF in drug discovery and disease:a diamond in the rough or split personality disorder[J].Expert Opin Drug Discov,2013,8(7):769-792.
• [7]Schilling T,Noth U,Klein-hitpass L,et al.Plasticity in adipogenesis and osteogenesis of human mesenchymal stem cells[J].Mol Cell Endocrinol,2007,271(1-2):1-17.
• [8]祝兆波,魏波.脂质促进成骨细胞凋亡及作用机制的研究进展[J].中国矫形外科杂志,2015,21(1):52-54.
• [9]Seibel MJ,Cooper MS,Zhou H.Glucocorticoid-induced osteoporosis:mechanisms,management,and future perspectives[J].Lancet Diabetes Endocrinol,2013,1(1):59-70.
• [10]Lecka-czernik B,Gubrij I,Moerman EJ,et al.Inhibition of Osf2/Cbfa1 expression and terminal osteoblast differentiation by PPARgamma2[J].J Cell Biochem,1999,74(3):357-371.
• [11]谭钢,罗磊,康鹏德,等.PPARγ-2和Runx-2与激素性股骨头坏死的发生[J].中国矫形外科杂志,2010,18(15):1274-1277.
• [12]Kim SW,Her SJ,Kim SY,et al.Ectopic overexpression of adipogenic transcription factors induces transdifferentiation of MC3T3-E1 osteoblasts[J].Biochem Biophys Res Commun,2005,2327(3):811-819.
• [13]Enomoto H,Furuichi T,Zanma A,et al.Runx2 deficiency in chondrocytes causes adipogenic changes in vitro[J].J Cell Sci,2004,117(3):417-425.