摘要
研究精氨酸血管加压素（AVP）对小鼠成骨细胞增殖和分化的影响及作用机制。（1）原代分离的小鼠成骨细胞中，加入100nmol／L的AVP培养72h后检测其对成骨细胞增殖作用的影响。（2）培养2、4、6、8、10d收集细胞培养液，并收集培养第10天的细胞裂解液，检测AVP对碱性磷酸酶（ALP）分泌的动态影响及细胞内ALP含量变化。（3）利用茜素红染色法检测AVP对培养8d和20d的成骨细胞形成钙结节的影响。（4）AVP作用20min，裂解细胞，利用放免法检测AVP对成骨细胞内cAMP的影响。结果显示，与对照组相比较，100nmol／L能够促进小鼠原代成骨细胞增殖（P〈0．01）。AVP能够使ALP的分泌量随着时间推移有显著增加（P〈0．01）；并增加成骨细胞形成钙结节数量和面积（P〈0．01）。同时AVP能够促进胞内cAMP水平的增加（P〈0．01）。结果提示AVP可能通过cAMP信号通路促进小鼠成骨细胞的增殖与分化。
The role of arginine vasopressin （AVP） played in proliferation and differentiation of mouse primary osteoblast and its mechanism was investigated. 100 nmol/ L AVP was added into the medium containing primary mouse osteoblast： （ 1 ） After being cultured for 72 h, the proliferation of the cells was counted with a cell counter. （ 2 ） The media of cultured cells on 2,4,6,8, 10 days were harvested and tested for the secreted ALP concentration by osteoblasts, and the cells were lysed in order to test the ALP concentration in cytosol. （ 3 ） The alizarin red staining was employed to detect the effect of AVP on calcium nodules formation on 8 th and 20 th days. （ 4 ） The osteoblast cells were incubated with AVP for 20 min, and then were lysed. Radioimmune assay was applied to test the change of cAMP in cytosol. These results showed that, compared to negative group, 100 nmol/L AVP significantly promoted the proliferation of primary mouse osteoblast （ P〈0.01 ）. ALP secretion was increased remarkably （ P〈 0.01 ）, and the number and area of calcium nodules were increased considerably （ P〈0.01 ）. The intracellular cAMP was increased after incubating cells with AVP for 20min （ P〈0.01 ）. These results suggest that AVP may promote proliferation and differentiation of mouse primary osteoblasts by cAMP signal pathway.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2015年第7期618-620,共3页
Chinese Journal of Endocrinology and Metabolism
基金
国家自然基金资助项目（31400651）
黑龙江省自然科学基金资助项目（D201067）
黑龙江省留学归国自然基金资助项目（LC201416）
关键词
精氨酸血管加压素
成骨细胞
增殖
分化
Arginine vasopressin
Osteoblast
Proliferation
Differentiation
作者简介
通信作者：侯霞。Email：hou_xia@hotmail．com
