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[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。
[Objective] To investigate the effect of PEDF on osteoblast migration and differentiation induced by high concentration of dexamethasone and explore the protective effect of PEDF on osteoblast function. [Methods] The MC3T3-E1 cells cultured in vitro were divided into six groups, which were control group, DEX 10-6mol / L dexamethasone group, PEDF dexamethasone group, PEDF dexamethasone group Groups were divided into three groups (2, 10, 50 nmol / L) and osteogenic induction group according to the concentration of PEDF. Transwell chamber and scratch experiments were used to observe the migration of cells after 48 h. The changes of cell morphology were observed by inverted phase contrast microscopy at 14 days. The changes of cell morphology were observed by cytochemical staining (alizarin red staining and oil red O staining). The expression of PPARγ-2 and Runx2 protein was detected by Western Blot. [Result] Under high concentration of dexamethasone, cell migration was inhibited, cell differentiation ability was weakened and lipid droplets were found in the cytoplasm. Western Blot showed that adipocyte marker protein PPARγ-2 was up-regulated, osteoblast marker Runx2 Decreased expression. PEDF intervention significantly reversed the effect of dexamethasone at high concentration on cell migration and differentiation. [Conclusion] High-dose glucocorticoid may inhibit osteoblast migration and osteogenic differentiation, promote osteoblast transdifferentiation into adipocytes, and PEDF can effectively reverse this process and play a role in protecting cell function.