中药Liu-OGN对体外成人成骨细胞增殖和分化的影响  

The Effects of Chinese Traditional Medicine Liu-OGN on Proliferation and Differentiation of Adult Human Osteoblasts in Vitro

作　　者：杨林[1] 陶天遵[1] 张淑云[2] 兰青 柳海峰 王新婷[1] 王晓亚 吴丽萍[1] 陶树清[1] 田军[1] 

机构地区：[1]哈尔滨医科大学附属第二医院骨科,哈尔滨150086 [2]哈尔滨医科大学附属第二医院科研实验中心 [3]哈尔滨市第五医院 [4]首都骨伤病研究所 [5]哈尔滨医科大学保健室

出　　处：《中国中医骨伤科杂志》2002年第1期1-3,共3页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：黑龙江省中医药管理局重点课题

摘　　要：目的 :研究不同浓度中药 Liu-OGN对体外成人成骨细胞增殖和分化的影响。方法 :采用胶原 -胰蛋白酶消化法获得成人松质骨中的成骨细胞 ,并进行纯化和培养 ,然后使用含不同浓度 Liu-OGN药物血清的培养液进行培养。通过倒置相差显微镜和电镜观察细胞生长情况 ,采用 MTT法检测细胞增殖情况 ,通过碱性磷酸酶和骨钙素等生化指标测定细胞分化情况。结果 :获得的成人成骨细胞生长情况良好 ,生化指标稳定可靠 ,纯化效果佳 ,可以满足相关研究的需要。进一步的研究结果表明 ,在成骨细胞密度为 1 0 0 0 0 /ml的条件下 ,与不加药对照组比较 ,1 0 %、2 0 % Liu-OGN中药组和地塞米松组的成骨细胞体积增大 ,上清液中碱性磷酸酶和骨钙素含量均明显增高 ;但各 Liu-OGN中药组成骨细胞的增殖情况无明显变化 ;结论 :中药 Liu-OGN对体外成人成骨细胞的分化有明显地促进作用 ,可以作为促进骨形成的药物使用。Objective:To observe and analyze the effects of Chinese traditional medicine Liu-OGN on proliferation and differentiation of adult human osteoblasts in vitro.Methods:Iliac trabecular bone specimens were obtained from the adult patients undergoing necessary surgery. After the bone pieces were digested with collagenase-trypsin, lots of osteoblasts were released and they were incubated at 37℃ in 95% relative humidity and 5% CO 2. Then the cells were purified and their passages were fed with DMEM-F12. Subsequently, the rat's serum containing Liu-OGN was added into the culture medium to incubate osteoblasts for three days and the control groups were incubated at the same time. All the cells were observed continually with light microscope and transmission electron microscope. Finally, cell proliferation was detected by the use of MTT method. Biochemical indexes, e.g., alkaline phosphatase (ALP) and osteocalcin (OCN), were determined to observe if there was any effect of Liu-OGN on differentiation of adult osteoblasts in vitro.Results:Of the adult osteoblasts obtained by collagenase-trypsin digestion, the survival was good, the biochemical indexes were stable, the degree of purification was excellent, and they were fit for the designed study. The following data demonstrated that under the condition of Liu-OGN serum at the concentration of 10% or 20% and osteoblasts 10000 /ml, there were significant promotion of ALP and OCN secretion without cell proliferation inducement.Conclusion:Chinese traditional medicine Liu-OGN at certain concentration has significant promoting effect on differentiation of adult human osteoblasts in vitro without obvious prolifer-ation inducement and can be used as medicine for bone formation.

关 键 词：中药 成骨细胞 增殖 分化 

分 类 号：R285.5[医药卫生—中药学]