色素上皮衍生因子对高浓度地塞米松刺激下成骨细胞迁移、分化的影响

发布时间：2018-10-26 11:05

【摘要】：[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。
[Abstract]:[objective] to study the effects of pigment epithelium-derived factor (pigment epithelium-derived factor,PEDF) on the migration and differentiation of osteoblasts cultured with high concentration of dexamethasone (Dexamethasone), and to explore the protective effect of PEDF on the function of osteoblasts. [methods] the MC3T3-E1 cells cultured in vitro were divided into six groups: control group, high concentration dexamethasone (dexamethasone,DEX 10-6mol/L) group and PEDF intervention dexamethasone group. The dexamethasone group treated with PEDF was divided into 3 groups according to the concentration of PEDF (2C1050 nmol/L) and osteogenic culture group. Cell migration was observed by Transwell chamber and scratch test 48 h later. The morphologic changes of the cells were observed by inverted phase contrast microscope at day 14. The expression of PPAR 纬 -2rRunx2 protein was detected by Western Blot. The cells were stained with alizarin red and oil red O. [results] under the action of high concentration of dexamethasone, cell migration ability was inhibited, cell differentiation ability was weakened, lipid drop, Western Blot appeared in the cytoplasm, the results showed that the expression of adipocyte marker PPAR 纬 -2 was increased. The expression of osteoblast marker protein Runx2 was decreased. PEDF treatment could reverse the inhibitory effect of high concentration dexamethasone on cell migration and differentiation. [conclusion] High dose of glucocorticoid may inhibit the migration and osteogenic differentiation of osteoblasts and promote the transformation of osteoblasts into adipocytes. PEDF can effectively reverse this process and protect the function of osteoblasts.
【作者单位】：徐州医科大学附属医院骨科;徐州医科大学研究生学院;徐州医科大学神经生物学实验室;
【基金】：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)
【分类号】：R580

【相似文献】