更年春方对破骨细胞的分化与凋亡及骨吸收活性的影响

Effects of "Gengnianchun Decoction" on Differentiation and Apoptosis and Activity of Bone Resorption of Osteoclasts

目的探讨更年春方对破骨细胞的分化、凋亡及骨吸收活性的影响。方法分离培养大鼠破骨细胞,加入含不同浓度的"更年春"药物血清培养液培养,生理盐水作为阴性对照组,尼尔雌醇作为阳性对照组,通过抗酒石酸酸性磷酸酶(TRAP)染色对TRAP阳性多核破骨细胞计数;用显微摄影结合计算机图像分析测定骨吸收造成的陷窝面积;采用Annexin V-FITC凋亡荧光染色结合TRAP染色法计数破骨细胞的凋亡率。结果与生理盐水组相比,尼尔雌醇组和更年春高剂量组TRAP染色阳性的破骨细胞数量显著减少(P<0.01);破骨细胞的凋亡率明显增加(P<0.01);尼尔雌醇组,更年春高、中剂量组骨吸收陷窝的面积明显减少(P<0.01)。与尼尔雌醇组相比,更年春高剂量组差别无显著性。结论更年春可抑制破骨细胞的形成和分化及骨吸收活性,促进破骨细胞的凋亡,从而发挥其抗骨质疏松的作用。

Objective To explore the effects of ＂Gengnianchun Decoction＇（GNC） on differentiation, activity of bone resorption and apoptosis of osteoclasts. Methods The osteoclasts were isolated from newborn rats, and treated with different culture media. Different concentrations of ＂Gengnianchun Decoction＂： high, moderate and low level groups（H group, M group and L group）, nilestriol （E group）, saline （N group） and sham （S group）. The numbers of tartrate resistant-acid phosphatase- positive [TRAP （＋）] multinucleated cells were counted by microscope. The surface area of bone resorption pits were measured by photomicrography and image analysis to evaluate the bone-resorbing activity of rat mature osteoclasts. The ratio of osteoclast in apoptosis was investigated through Annexin V-FITC apoptosis detection combined with TRAP staining. Results The number of TRAP （＋） osteoclasts was lower and the ratio of apoptosis osteoclasts was higher in H and E groups than those of N group （P〈0.01）, H, E and M groups significantly decreased the area of bone resorption pits, as compared with N group （P〈0.01）. The effect between H group and E group did not show significant differences. Conclusion The mechanism of GNC in preventing osteoporosis may be related with its inhibiting osteoclasts formation, differentiation, and activity of bone resorption, stimulating apoptosis of osteoclasts.