染料木黄酮对人成骨细胞凋亡的影响

目的: 观察染料木黄酮(Gen)对人成骨细胞凋亡的影响。方法: 将体外分离到的人骨膜成骨细胞在高糖DMEM(含10 %小牛血清)中进行纯化扩增至第三代,然后接种于无酚红高糖DMEM培养液中(含10 %经活性碳-葡聚糖苷处理的胎牛血清,CDT-FBS),4 d后用0.25 %的胰蛋白酶消化、收集细胞,用PBS洗涤2次后,用于各项实验。指标包括:流式细胞仪分析细胞凋亡比例,琼脂糖凝胶电泳检测DNA断裂条带,免疫组化观察Gen对Bax、Bcl-2及caspase-3表达的影响。结果: 无雌激素(E2)血清培养可诱导成骨细胞发生凋亡(45.7 %),加入10-8 mol/L E2、10-6 mol/L 和10-5 mol/L Gen对细胞处理48 h,成骨细胞凋亡率分别降低到15.3 %、36.3 %及28.4 %(P<0.05)免疫组化结果表明,同E2类似,Gen可抑制Bax和capsase-3的表达,而促进Bcl-2蛋白的表达。结论: 染料木黄酮可能是通过影响雌激素受体途径而发挥其骨保护作用的。

EFFECTS OF GENISTEIN ON APOPTOSIS IN HUMAN OSTEOBLAST CELLS

Objective:To investigate the effects of genistein (Gen) on apoptosis in human osteoblast. Methods:Human periosteum was maintained in DMEM medium containing 10% newborn calf serum through four passages to isolate and proliferate osteoblast. Then, cells were maintained in phenol red-free DMEM supplemented with 5% charcoal-stripped FCS for 4 d before addition of treatments. Flow cytometer and DNA fragment were used to explore apoptosis in osteoblast, and immunohistochemistry was used to study protein expression of Bax, Bcl-2 and caspase-3. Results:Estrogen depletion could induce apoptosis in human osteoblast. With the similarity of 10-8 mol/L E2, at the conentrations of 10-6 mol/L and 10-5 mol/L, Gen could inhibit apoptosis induced by estrogen depletion. Compared with vehicle control, Gen significantly decreased protein expression of Bax and caspase-3 and increased protein expression of Bcl-2. Conclusion:Genistein could inhibit apoptosis induced by estrogen depletion in human osteoblast.