目的探讨丹皮酚拮抗过氧亚硝基阴离子(ONOO-)对体外培养大鼠成骨细胞凋亡的作用。方法用改良的组织块法分离培养新生大鼠颅骨成骨细胞,采用淬灭流动反应方法体外制备ONOO-,以1 mmol/L终浓度加入成骨细胞培养体系,作用30 min后弃去,继续常规培养12 h,用Hoechst33258染色法及TUNEL法检测成骨细胞的凋亡,并以0.1 mmol/L丹皮酚消除ONOO-(1 mmol/L)对大鼠成骨细胞凋亡的影响。结果1 mmol/L的ONOO-可导致大鼠成骨细胞凋亡;0.1 mmol/L丹皮酚能拮抗ONOO-(1 mmol/L)所引起的大鼠成骨细胞凋亡。结论丹皮酚0.1 mmol/L在体外具有拮抗ONOO-(1 mmol/L)致大鼠成骨细胞凋亡的作用。

Objective To investigate the effect of paeonol antagonizing peroxynitrite(ONOO-)0n osteoblast apoptosis of rat in vitro.Methods Osteoblasts of newly born rat’S skull were separated and cultured by tissue mass method.ONOO-was produced by chemical synthesis with a quenched flow reactor.ONOO-at 1 mmol/L concentration was added to cultured osteoblast system in vitro.After incubated for 30 min,osteoblasts were continued to be cultured for 12 h with routine method.Hoechst33258 stain and TUNEL method were used to detect the osteoblast apoptosis and 0.1 mmol/L concentration of paeonol was used to eliminate the effect of ONOO-(1mmol/L)on osteoblast apoptosis of rat.Results ONOO-(1 mmol/L)could lead to rat osteohlast apoptosis.Paeonol of 0.1 mmol/L concentration could antagonize rat osteoblast apoptosis induced by ON00-(1 mmol/L).Conclusion Paeonol(0.1 mmol/L)has the significant antagonizing effect on ONOO-of rat osteoblast apoptosis in vitro.

河北省卫生厅医学科学重点资助项目(04062);河北省教育厅博士基金资助项目(B2004122);石家庄市科技攻关项目(04146173A);河北医科大学博士基金资助项目(040028)