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目的:探讨持续性压力促进成骨细胞分泌血管内皮生长因子(VEGF)的机制。方法:取1～2日龄SD大鼠颅盖骨进行成骨细胞原代培养,检测并鉴定成骨细胞。细胞培养至第3代,分为加压组和不加压组,每组再分成PD98059不预处理组和预处理组。加压组在密闭容器内采用压缩空气施以100 kPa的静压力,分别加压0.5、2.0、6.0 h,ELISA法检测培养液中VEGF浓度,RT-PCR检测VEGF mRNA的变化;Western blot检测各组成骨细胞内磷酸化ERK1/2(pERK1/2)的水平。结果:持续性压力促进成骨细胞VEGF mRNA的表达和蛋白的分泌,同时也明显增加ERK1/2的磷酸化水平;而ERK1/2总蛋白的量却无明显变化。PD98059在显著抑制持续性加压所诱导的成骨细内ERK1/2磷酸化水平的同时,抑制了VEGF的表达及分泌。结论:持续性压力通过ERK1/2的激活调节成骨细胞VEGF的分泌。
Objective: To investigate the mechanism of sustained pressure on osteoblasts to secrete vascular endothelial growth factor (VEGF). Methods: Primary osteoblasts were obtained from calvaria of 1 to 2 days old SD rats, and osteoblasts were detected and identified. The cells were cultured to the third generation and divided into two groups: pressurized group and non-pressurized group. Each group was divided into PD98059 non-pretreatment group and pre-treatment group. Pressure group in a sealed container with compressed air at 100 kPa static pressure, respectively, pressure 0.5,2.0,6.0 h, VEGF concentration in culture medium was detected by ELISA, VEGF mRNA was detected by RT-PCR; Western blot detection Constitutes the level of phosphorylated ERK1 / 2 (pERK1 / 2) in osteocytes. Results: Sustained pressure promoted the expression of VEGF mRNA and protein in osteoblasts, but also significantly increased the phosphorylation of ERK1 / 2. However, the amount of total ERK1 / 2 protein did not change significantly. PD98059 inhibited the expression and secretion of VEGF while significantly inhibiting the phosphorylation of ERK1 / 2 induced by sustained pressurization. CONCLUSION: Sustained stress regulates the secretion of VEGF from osteoblasts through ERK1 / 2 activation.