异补骨脂素对体外培养大鼠成骨细胞增殖分化成熟的影响  

Effect of Isopsoralen on the Proliferation and Differentiate of Osteoblasts in vitro

作　　者：明磊国[1] 陈克明[1] 葛宝丰[1] 马慧萍[1] 翟远坤[1] 

机构地区：[1]兰州军区总医院,甘肃兰州730050

出　　处：《中药材》2011年第3期404-408,共5页Journal of Chinese Medicinal Materials

摘　　要：目的:研究异补骨脂素(Isopsoralen,IP)对体外培养大鼠颅骨成骨细胞(rat calvarial osteoblasts,ROB)增殖与分化成熟的影响。方法:取新生SD大鼠颅骨,多次酶消化法获得成骨细胞,培养于含10%FBS的MEM培养液中,3 d后首次换液,铺满90%皿底后传代培养;增殖分析采用96孔板,加入在培养液中终浓度分别为1×10-4、1×10-5、1×10-6、1×10-7mol/L的异补骨脂素,MTT法分析;分化分析采用24孔板,于成骨性诱导培养第3、6、9、12、15天分别测碱性磷酸酶(Alkaline Phosphatase,ALP)活性、钙含量、骨钙素,第12天进行ALP组织化学染色,第14天进行茜素红染色和钙化结节计数。结果:终浓度为1×10-4mol/L的异补骨脂素对细胞增殖有抑制作用,而1×10-5mol/L浓度虽对ROB增殖无明显影响,但能显著提高细胞ALP活性、增加钙含量、促进骨钙素分泌,并增加钙化结节数量。结论1×10-5mol/L异补骨脂素能促进ROB分化成熟,应为中药补骨脂抗骨质疏松的有效成分。Objective：To investigate the effect of isopsoralen on proliferation,osteogenic differentiation and calcification capacity of rat calvarial osteoblasts（ROB）.Methods： Segregated neonatal SD rat skull,and digestion with enzyme to obtain bone cells and cultured in MEM containing 10% FBS.Exchange the medium after three days,proceeded serial subcultivation when cells covered with 90% culture dish.Proliferation analysis was performed in 96-well plates use MTT method,isopsoralen′s final concentration were 1×10-4,1×10-5,1×10-6,1×10-7 mmol/L.Differentiation analysis was performed in 24-well plates,the Alkaline phosphatase activity and calcium salt sediment yield and osteocalcin measured at the 4th,8th,12th,16th day.At 12th day,proceeded ALP stain,and at 14th day for alizarin red staining and calcified nodule count.Results： When the Isopsoralen′s final concentration was 1×10-5mmol/L,there was no significant effect on the ROB′s proliferation,but it could promote osteogenesis.It also could raise the ALP activity and calcium salt sediment yield and osteocalcin,increase calcified tubercle amount.Conclusion： When the isopsoralen final concentration is 1×10-5mmol/L,it promoted ROB differentiation and maturation.Isopsoralen may be the active ingredients of preventing anti-osteoporosis in Psoralea corylifolia.

关 键 词：成骨细胞 细胞培养 异补骨脂素 骨质疏松 增殖 分化 

分 类 号：R285.5[医药卫生—中药学]