BMP-2联合雷奈酸锶对大鼠成骨细胞增殖和分化的影响

Effects of BMP-2 combined with Strontium ranelate on proliferation and differentiation of rat osteoblasts

目的探讨骨形成蛋白(BMP)-2联合雷奈酸锶(SR)对大鼠成骨细胞增殖及分化的影响。方法 2018年3月以胰酶-胶原酶消化法分离获取大鼠原代成骨细胞,随机分4组,骨形成蛋白组加入0. 1 mg/L的BMP-2;雷奈酸锶组加入5 mmol/L的SR;联合组:先加入0. 1 mg/L的BMP-2,培养6 d换液后再次加入5 mmol/L的SR;空白对照组不加入任何诱导液,培养12 d。茜素红染色、碱性磷酸酶(ALP)活性观察成骨细胞功能状态,成骨细胞增殖测定,对骨钙素(OCN)、Runx2蛋白表达测定。结果骨形成蛋白组、雷奈酸锶组、联合组较空白对照组ALP活性含量、大鼠成骨细胞数目增加、OCN、Runx2表达含量增高,其中以联合组最高,骨形成蛋白组、雷奈酸锶组两组ALP活性含量、大鼠成骨细胞数目增加、OCN、Runx2表达含量差异无显著性(P> 0. 05)。结论 BMP-2与SR联合应用较单独应用更能促进大鼠成骨细胞增殖及分化。

Objective To investigate the effect of BMP - 2 combined with SR on the proliferation and differentiation of rat osteoblasts. Methods In March 2018, rat primary osteoblasts obtained by trypsin - collagenase digestion were randomly divided into 4 groups： 0.1 mg/L BMP - 2 in the bone morphogenetic group ; 5 mmol/in the strontium ranelate group. SR of L; combination group ： first added 0.1 mg/L BMP - 2, cultured for 6 d, and then added 5 mmol/L SR again after changing the solution ; blank control group did not add any induction solution, cultured for 12 d. Alizarin red staining, ALP activity to observe osteoblast functional status, osteoblast proliferation assay, OCN and Runx2 protein expression assay. Results The content of ALP in the bone morphogenetic group, the strontium ranelate group and the combined group was higher than that in the blank control group, the number of osteoblasts increased, and the expression of OCN and Runx2 increased. Among them, the combined group was the highest. There was no significant difference in the ALP activity, the number of osteoblasts and the expression of OCN and Runx2 between the bone morphogenetic protein group and the ranitonic acid group （ P 〉 0.05）. Conclusion The combination of BMP - 2 and SR can promote the proliferation and differentiation of rat osteoblasts compared with the single application.