摘要
目的观察p38MAPK信号转导通路在成骨细胞分化及核因子-κB受体激活配体(receptor activator of nuclear factor-κBligand,RANKL)和骨保护素(osteoprotegerin,OPG)表达中的作用。方法取第1继代BALB/c小鼠颅盖骨成骨细胞,药物刺激组分别加入10-8mol/L 17β-雌二醇和10-7mol/L雷洛昔芬;含阻断剂组预先添加5μmol/L SB202190阻断p38MAPK通路后,再加17β-雌二醇或雷洛昔芬。72 h后用PNPP法测定成骨细胞内碱性磷酸酶(alkaliphosphatase,ALP)活性;RT-PCR法检测成骨细胞ALP、OPG和RANKL的转录水平。结果17β-雌二醇和雷洛昔芬能够促进成骨细胞分化,促进OPG、RANKL的表达(P<0.05),OPG/RANKL无明显变化(P>0.05);阻断p38MAPK信号转导通路后,成骨细胞分化受抑,OPG、RANKL的表达和OPG/RANKL降低(P<0.05)。结论p38MAPK抑制可干扰体外培养成骨细胞分化,抑制RANKL和OPG的过度表达。
Objective To investigate the role of p38MAPK in the differentiation of murine osteoblasts,and to observe the expressions of receptor activator of nuclear factor-κB ligand（RANKL） and osteoprotegerin（OPG）. Methods The calvarial osteoblasts of newborn BALB/c mice were cultured in MEM medium containing 10% NCS.Raloxifene（10^-7 mol/L） and 17β-estradiol（10^-8 mol/L） were added respectively when cells reached 70%-80% confluence combined with or without 5 μmol/L SB202190,an inhibitor of p38MAPK.The osteoblasts alkaline phosphatase activity assays were performed 72 hours later using PNPP method,and mRNA levels of alkaliphosphatase（ALP）,OPG and RANKL were determined by RT-PCR. Results 17β-estradiol and raloxifene increased ALP activity and ALP mRNA level in osteoblasts in vitro which were blocked by p38MAPK inhibitor.The mRNA levels of RANKL and OPG were up-regulated by 17β-estradiol and raloxifene while the ratio of OPG/RANKL kept constant.SB202190（5 μmol/L） inhibited the highly expressed RANKL and OPG in osteoblasts,and obviously decreased the ratio of OPG/RANKL. Conclusions p38MAPK inhibition blocked the differentiation of osteoblasts and decreased the up-regulated OPG and RANKL expressions in osteoblasts significantly（P〈0.05）.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2010年第1期39-42,共4页
Fudan University Journal of Medical Sciences
基金
上海市医药重点学科建设基金资助项目(5Ⅲ016)
