二仙汤含药血清对过氧化氢诱导成骨细胞骨形成的影响  

Effect of Serum Containing Erxian Decoction(二仙汤) on Bone Formation of Osteoblasts Induced by Hydrogen Peroxide

作　　者：刘波[1] 吴琪[1] 刘志文[1] 徐彭[1] 杨硕[1] 谢慧慧[1] 王莹[1] LIU Bo;WU Qi;LIU Zhiwen;XU Peng;YANG Shuo;XIE Huihui;WANG Ying(College of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 33000)

机构地区：[1]江西中医药大学药学院

出　　处：《中医杂志》2018年第15期1318-1322,共5页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81560723);江西省自然科学基金计划(20151BAB205072)

摘　　要：目的探讨二仙汤治疗骨质疏松症的可能作用机制。方法成骨细胞以5×103/孔接种于96孔板,分为正常组、模型组、戊酸雌二醇组和二仙汤高、中、低剂量组,每组6孔。除正常组外其余各组用含600μmol/L过氧化氢(H2O2)的完全培养基培养3h诱导成骨细胞骨形成模型。正常组用空白对照血清培养,戊酸雌二醇组给予8%戊酸雌二醇含药血清培养,二仙汤高、中、低剂量组分别给予8%、4%、2%二仙汤含药血清培养48h,检测成骨细胞增殖、碱性磷酸酶(ALP)活力、钙结节生成以及骨形成蛋白2(BMP-2)和骨保护素(OPG)蛋白表达。另外以钙离子通道阻断剂Nifdipine(10μmol/L)进行干预,观察Nifdipine对H2O2刺激成骨细胞后细胞增殖、ALP活性及BMP-2、OPG蛋白表达以及Nifdipine干预下二仙汤含药血清对H2O2刺激成骨细胞后细胞增殖、ALP活性及BMP-2、OPG蛋白表达的影响。结果与正常组比较,模型组细胞增殖、ALP活性、钙结节及BMP-2表达降低(P<0.05或P<0.01)。二仙汤各剂量组能不同程度促进成骨细胞增殖,提高ALP活力和钙结节生成,促进BMP-2、OPG蛋白表达(P<0.05或P<0.01)。Nifdipine能提高H2O2刺激下成骨细胞BMP-2、OPG蛋白表达(P<0.05或P<0.01)。与Nifdipine组比较,二仙汤含药血清和Nifdipine合用则显著提高H2O2刺激下的成骨细胞增殖,降低BMP-2、OPG蛋白表达(P<0.01)。结论二仙汤含药血清能明显改善氧化刺激对成骨细胞骨形成的影响,其机制可能与调节钙离子通道有关。Objective To explore the possible action mechanism of Erxian Decoction（ 二仙汤） in the treatment of osteoporosis. Methods Osteoblasts were inoculated into 96-well plates at 5 × 10^3 holes and divided into normal group,model group,estradiol valerate group and Erxian Decoction high,medium and low dose groups,with 6 holes in each group. All groups except the normal group,were cultured in complete medium containing 600 μmol/L hydrogen peroxide（ H2O2） for 3 h to induce osteoblast formation model. The normal group was cultured with blank control serum. The estradiol valerate group was given serum containing 8% valerate estradiol,and the Erxian Decoction high,medium and low dose groups were given 8%,4%,2% Erxian Decoction respectively. The drug-containing serum was cultured for 48 h. The osteoblast proliferation,alkaline phosphatase（ ALP） activity,calcium nodule formation,and the expression of bone morphogenetic protein 2（ BMP-2） and osteoprotegerin（ OPG） protein were detected. In addition,the intervention of Nifdipine（ 10 μmol/L） was used to observe the effects of Nifdipine on cell proliferation,ALP activity,BMP-2 and OPG protein expression after H2 O2 induced osteoblasts and its effects under the intervention of Nifdipine. Results Compared with the normal group,cell proliferation,ALP activity,calcium nodules and BMP-2 expression were decreased in the model group（ P〈0. 05 or P〈0. 01）. Groups with different doses of Erxian Decoction could promote osteoblast proliferation,increase ALP activity and calcium nodule formation,and promote BMP-2 and OPG protein expression（ P〈0. 05 or P〈0. 01）. Nifdipine could increase BMP-2 and OPG protein expression of osteoblasts induced by H2 O2（ P〈0. 05 or P 〈0. 01）. The combination of serum containing Erxian Decoction and Nifdipine could significantly increase the proliferation of osteoblasts induced by H2 O2 and decrease the expression of BMP-2 and OPG protein as compared with simple use of Nifdipine（ P〈0. 01）. Conclusion The se

关 键 词：骨质疏松症 二仙汤 成骨细胞 过氧化氢 钙离子通道 

分 类 号：R285[医药卫生—中药学]