摘要
从牛骨制备低分子质量骨多肽,并观察其对大鼠破骨细胞的影响。采用冷冻离心、等电点沉淀方法去除骨胶原,用超滤法得到分子质量10000u以下的牛骨多肽,并用Sephadex G-10层析脱盐。Tricine-SDS-PAGE对低分子质量牛骨多肽分子质量进行检测,采用抗酒石酸磷酸酶(TRAP)染色法研究牛骨多肽对体外培养大鼠破骨细胞的分化影响,甲苯胺蓝硼酸盐染色法观察对破骨细胞的骨吸收影响以及HE、TRAP、DAPI染色法分别观察凋亡征象。结果:建立和优化了牛骨多肽的制备方法;牛骨多肽对破骨细胞分化无影响,但能抑制其骨吸收功能,促进其凋亡。
To prepare low molecular mass polypeptides from bovine bone and to study the effects on osteoclast of rat, degenerative collagen from bovine bone solution by bypotbermy centrifugalization and isoelectric point precipitation was removed, polypeptides from bovine bone which molecular mass was below 10kD by uhrafihration was obtained, and desalted by Sephadex （3-10 chromatography. The molecular quality of low molecular mass polypeptides was detected by Tricine-SDS-PAGE electrophoresis, the differentiation of osteoclast was studied and tested by TRAP staining and bone resorption of osteoclast by toluidine blue borate staining, and apoptosis of osteoclast by HE, TRAP, DAP1 staining at the low molecular mass polypeptides action. The preparation of low molecular mass polypeptides from bovine bone is established and optimized. The polypeptides have no effect on differentiation of osteoclast but they inhibit bone resorption of osteoclast and promote apoptosis of osteoclast.
出处
《药物生物技术》
CAS
CSCD
2007年第1期51-55,共5页
Pharmaceutical Biotechnology
基金
校企横向联合项目2005001
关键词
牛骨多肽
制备
破骨细胞
凋亡
Bovine bone, Low molecular mass polypeptides, Preparation, Osteoclast, Apoptosis
