晚期氧化蛋白产物对大鼠牙槽骨细胞凋亡的影响

目的 探讨晚期氧化蛋白产物(AOPP)对大鼠牙槽骨细胞凋亡的诱导作用.方法 选取大鼠牙槽骨细胞作为靶细胞,用不同浓度(0、100、200 μg/mL)的AOPP作用不同时间(12、24 h)刺激牙槽骨细胞,通过流式细胞仪测定细胞凋亡情况,酶标仪测定刺激下牙槽骨细胞活性氧(ROS)的生成量,采用免疫印迹法检测影响细胞凋亡的相关蛋白Bcl-2和Bax的表达变化.结果 原代培养获得的牙槽骨细胞呈贴壁生长,能合成碱性磷酸酶,具有成骨样细胞表现.AOPP可以引起牙槽骨细胞产生大量ROS,引起细胞凋亡,随着作用剂量的增大和作用时间的延长,细胞凋亡率明显升高,其中当AOPP作用浓度为200 μg/mL、作用时间为24 h时,凋亡率最高.牙槽骨细胞可表现为Bcl-2合成减少,Bax合成增加,而且相关凋亡因子表达随着作用时间的延长和作用浓度的增加而显著升高(P<0.05).结论 氧化应激可引起牙槽骨细胞凋亡,这可能是导致部分口腔慢性炎症性疾病后期出现牙槽骨吸收的原因.

Effects of advanced oxidative protein products on the apoptosis of rat alveolar osteoblasts

Objective To investigate the mechanism of advanced oxidative protein product (AOPP)-induced apoptosis of alveolar osteoblasts in rats.Methods AOPP were used to stimulate rat alveolar bone cells.The cell apoptosis ratio was evaluated by flow cytometry.The reactive oxygen species (ROS) produced by the alveolar osteoblasts were measured by ELISA.Western blot was used to detect the expression of the apoptosis related proteins in the alveolar osteoblasts, including Bcl-2 and Bax.Results The primary alveolar bone cells were shown osteogenic-like potential.AOPP induced production of large amount of ROS in alveolar bone cells and resulted in cell apoptosis.The cell apoptosis ratio was significantly increased with the increase of AOPP (P<0.05).The expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated.On the other hand, the cell apoptosis ratio was significantly increased with the prolonging time of AOPP (P<0.05).When the concentration of AOPP reach at 200 μg/mL or the stimulating duration reached at 24 h, the apoptosis rate was the highest.Conclusion Oxidative stress can cause alveolar osteoblasts apoptosis, and causes alveolar bone absorption in the oral chronic inflammatory disease.