饲补肾中药大鼠血清对成骨细胞-破骨细胞共育系中破骨细胞功能的影响

目的探讨喂饲补肾中药大鼠血清在成骨细胞-破骨细胞培养体系中对破骨细胞骨吸收功能的影响。方法取SD大鼠的胎鼠颅骨与四肢骨分别分离、培养成骨细胞和破骨细胞,利用共育系使二者生活在同一环境中,施加高、中、低剂量的饲补肾中药大鼠血清,并与对照组及单独培养的破骨细胞进行比较,酶动力学法测定培养上清中抗酒石酸酸性磷酸酶(TRACP)的活性,利用甲苯胺蓝对骨吸收陷窝染色并在Leica Quantimet 500图像分析仪下测定骨吸收陷窝的面积和数目。结果共育系中对TRACP活力的抑制及骨陷窝面积与数目的减少以中剂量组作用最为明显(与对照组比较P<0.01),单独培养3个剂量组与对照组比较均有显著性差异(P<0.05),以中剂量组作用最为明显(P<0.01)。结论饲补肾中药大鼠血清可以抑制共育系中破骨细胞的骨吸收功能。

The Effects of Serum from Kidney-tonifying Traditional Chinese Drug-fed Rats on Osteoclasts in the Osteoblast-osteoclast Co-culture System

Objective To investigate the effects of serum from kidney-tonifing traditional Chinese drug-fed rats on the osteoclasts in the osteoblast-osteoclast co-culture system.Methods The osteoblasts and osteoclasts were isolated respectively from the skull and limb of SD rats and co-cultured in the same system,the culture media containing the serum from rats fed with the drugs at different doses(high,mediate and low doses) was used to treat the osteoblast-osteoclast co-culture system and the system by osteoclasts only.The tartrate resistant acid phosphatase(TRACP) activities in the medium were measured by enzyme kinetics.The resorption pits were stained by toludine blue.The area and number of resorption pits were determined with the Leica Quantimet 500 system.Results Under the mediate dose of serum from kidney-tonifying traditional Chinese drug-fed rats in the co-culture system,TRACP activities,the area and number of resorption pits was markedly restrained(vs control P<0.01);in the system by osteoclasts only,high,mediate and low doses had effects markedly(P<0.05) but the mediate doses were the best of them(P<0.01).Conclusion The function of osteoclast is restricted in the co-culture system in which serum from kidney-tonifying traditional Chinese drug-fed rats is administered.