目的 利用体外培养大鼠成骨细胞,研究δ阿片受体激活对血清饥饿诱导成骨细胞凋亡的影响并探讨其机制.方法 培养大鼠成骨细胞,分组给药作用48 h后,MTT法测定细胞存活率,流式细胞仪分析细胞周期,Annexin V-F1TC/PI双标记法测定细胞凋亡率,Western blot测定PKC和Caspase-3表达.结果 给予δ阿片受体激动剂DADLE l μmol·L-1处理可显著抑制由血清饥饿导致的成骨细胞凋亡,表现为细胞存活率增加,凋亡率降低,S+G2+M期百分比增加,Caspase-3表达下降,PKC表达增加；但这种现象可被Naltrindole所拮抗；并且给予10 μmol·L-1 GF109203X亦可拮抗DADLE的这种抑制细胞凋亡的作用,表现为细胞存活率降低,凋亡率增加；S+G2-M期百分比降低；caspase-3表达增加；PKC表达下降.结论 DADLE激活δ阿片受体对血清饥饿诱导的成骨细胞凋亡起保护作用,且这种作用是通过PKC途径实现的.%Objective To culture rat osteoblasts in vitro and study the effects of activated S opium receptor on apoptosis of rat osteoblasts induced by starvative blood serum and its mechanism. Methods The rat osteoblasts were cultured and administered activated S opium receptor . After 24 horns,the survival rate of the cells were detected by MTT Method;the cell cycle was analyzed by flow cytomeuy (FCM); the rate of cellular apoptosis was determined by the paired labeled technique of Annexin V-FTTC/PI; the expression of PKC and Caspase-3 were determined by Western blot. Results After the administration of 1 μmol·L-1 DADLE,an agonist of δ opium receptor,the apoptosis of osteoblasts induced by starvative blood serum was inhibited significantly. So,the survival rate of the cells increased and the rate of cellular apoptosis decreased;the percentage of S+G2+M raised;the expression of Caspase-3 decreased and the expression of PKC increased. However, the inhibition can be antagonised by Naltrindole and 10 μmol-L1 GF109203X. It is manifested as follows: the survival rate of the cell reduced and the apoptosis rat of the cell increased;the percentage of S+G2+M reduced;the expression of Caspase-3 increased and the expression of PKC decreased. Conclusion The apoptosis of osteoblasts induced by starvative blood serum can be protected by 8 opium receptor activated by DADLE,and this protection is achieved by PKC.
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