强骨宝方载药血清灭活与否对成骨细胞增殖功能的影响

目的：探讨强骨宝方糖尿病鼠血清灭活与否对成骨细胞增殖的影响。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，倒置显微镜下观察其形态，碱性磷酸酶（ALPase）染色法、钙化结节染色、Van—Gieson染色鉴定细胞后用灭活和不灭活的不同时效（灌胃3、5d，末次灌胃l、3h）、不同浓度（5％、10％、20％）的强骨宝方糖尿病模型鼠血清加入成骨细胞培养体系，作用一定的时间，应用MTT比色法检测载药血清灭活对成骨细胞增殖能力的影响。结果：强骨宝方灌胃3、5d，末次灌胃l、3h的5％、10％、20％糖尿病鼠不灭活血清与灭活组相比差异有统计学意义，以不灭活载药血清对成骨细胞增殖功能的促进作用较强（P〈0．01或P〈0．05）。结论：强骨宝方糖尿病模型鼠血清灭活与否影响成骨细胞的增殖功能，以不灭活载药血清的作用最佳。

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

Objective ：To study the effect of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction （强骨宝方 ）on the proliferation of osteoblast cells （OB）cultured in vitro. Methods： OB was isolated from the skull of newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion and identified by image analysis under inverted microscope, V-G collagen staining, ALP staining, calcification nod staining etc. After the OB was identified, in activated and un-inactivated pharmaco-serum of diabetic rats fed with Qianggubao decoction （ 强骨宝方 ） of different phase （rats were fed with medicine 3 days or 5 days after last fed with medicine 1 hour or 3 hours） and concentration （5%, 10% ,20% ） were added to the OB and incubated. After determined times,the effects of the proliferation of osteoblasts were detected by MTY analysis. Results：There was significant difference between un-inactivated pharmaco-serum and inactivated pharmaco-serum on the proliferation of osteoblasts, and un-inactivated serum had stronger effects to improve the proliferation of osteoblasts （P〈0.01 or P〈0.05 ）. Conclusion： Un-inactivated and inactivation pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction （ 强骨宝方 ）can influence the proliferation of, and the un-inactivated pharmaco-serum has stronger effects.