强骨宝方含药血清对糖尿病模型鼠体外培养成骨细胞的影响

目的：探讨糖尿病模型鼠强骨宝方含药血清促进体外培养成骨细胞分化、矿化的最佳时相与量效。方法：采用胰蛋白酶-Ⅱ型胶原酶消化法从1-2日龄SD大鼠颅盖骨中分离出成骨细胞，鉴定细胞后，用不灭活的不同时效（灌胃3、5d，末次灌胃后1、3h）、不同浓度（5％、10％、20％）的强骨宝方糖尿病模型鼠含药血清加入成骨细胞培养体系，培养7、18d，分别观察各组ALP活性及矿化结节形成量。结果：各浓度的糖尿病模型鼠强骨宝方含药血清对成骨细胞分泌ALP及矿化结节形成的作用均优于模型对照组，并接近于正常对照组水平，其中以20％添加浓度的作用最明显。在时相方面，两指标均以灌药3d或5d、末次灌胃后1h取得血清作用最明显。结论：糖尿病模型大鼠强骨宝方含药血清对体外培养成骨细胞（OB）的分化及矿化功能有促进作用。考虑到实验的时间与成本，以灌胃3d、末次灌胃后1h、20％不灭活的强骨宝方糖尿病模型鼠含药血清对成骨细胞分化与矿化作用最适宜。

Experimental study on the effects of pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction (强骨宝方) on osteoblast culture in vitro

Objective ：To investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction （强骨宝方）on the differentiation and mineralization of osteoblast （OB）. Methods ：OB was isolated from the skull of newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified ,different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction （强骨宝方）of different phase （rats were fed with medicine three days or five days after last fed with medicine one hour or three hours） and concentration （5%, 10%, 20% ）were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected. Results：The secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum ,the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction （ 强骨宝方）three days or five days,after one hour of last fed with medicine. Conclusion： The pharmaco-serum of diabetic rats fed with Qianggubao decoction （ 强骨宝方）can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction （强骨宝方）three days,after one hour of last fed,with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.