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Effects of pigment epithelium- derived factor on migration and differentiation of osteoblasts after stimulation by high concentrations of dexamethasone
孙少松;郑欣;姚圣城;褚耿磊;庞勇;董红燕;赵凤朝;郭开今;
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
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;
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