CAJ | 학술논문

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100 nmol/ L 的 AVP 培养72 h 后检测其对成骨细胞增殖作用的影响。(2)培养2、4、6、8、10 d 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8 d 和20 d 的成骨细胞形成钙结节的影响。(4)AVP 作用20 min,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,100 nmol/ 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。
연구정안산혈관가압소(AVP)대소서성골세포증식화분화적영향급작용궤제。(1)원대분리적소서성골세포중,가입100 nmol/ L 적 AVP 배양72 h 후검측기대성골세포증식작용적영향。(2)배양2、4、6、8、10 d 수집세포배양액,병수집배양제10천적세포렬해액,검측 AVP 대감성린산매(ALP)분비적동태영향급세포내 ALP 함량변화。(3)이용천소홍염색법검측 AVP 대배양8 d 화20 d 적성골세포형성개결절적영향。(4)AVP 작용20 min,렬해세포,이용방면법검측 AVP 대성골세포내 cAMP 적영향。결과현시,여대조조상비교,100 nmol/ L 능구촉진소서원대성골세포증식(P<0.01)。 AVP 능구사 ALP 적분비량수착시간추이유현저증가(P<0.01)；병증가성골세포형성개결절수량화면적(P<0.01)。동시 AVP 능구촉진포내 cAMP 수평적증가(P<0.01)。결과제시 AVP 가능통과 cAMP 신호통로촉진소서성골세포적증식여분화。
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.