益骨胶囊含药血清对共育体系中破骨细胞活性和凋亡的影响

目的： 观察益骨胶囊含药血清在成骨-破骨细胞共育体系中对SD大鼠破骨细胞（OC）活性和凋亡的影响。 方法： (1)取1d龄SD大鼠颅骨分离培养成骨细胞（OB），取5d龄SD大鼠四肢股骨、胫骨分离培养OC，建立细胞上清相通但细胞间不相互混杂的平面式成骨-破骨细胞共育体系，实验分为含药血清组和对照组；（2）将10月龄SD雌性大鼠分为益骨胶囊灌胃组和生理盐水对照组，制备含药血清和对照血清；（3）重氮盐法检测抗酒石酸酸性磷酸酶（TRAP）和光镜观察骨陷窝数；（4）光镜和荧光显微镜下观察共育体系中OC凋亡情况。 结果： 含药血清组在48 h、72 h、96 h对成骨-破骨细胞共育体系中OC分泌TRACP的活性均明显降低于对照组，OC的存活数明显低于对照组，OC的凋亡率明显高于对照组且呈明显的时效关系；所形成骨吸收陷窝的数目明显低于对照组(P<0.01)。 结论： 益骨胶囊含药血清在共育体系中能够抑制OC活性，诱导破骨细胞的凋亡。

Effect of BBC-containing serum on the activity and apoptosis of osteoclasts in osteoblasts and osteoclasts co-culture system

AIM: To study the effect of Bone-benefiting capsule(BBC)-containing serum on activity and apoptosis of osteoclasts(OC) in osteoblasts(OB) and osteoclasts co-culture system.METHODS:(1) OB were separated from the skull of SD rats aged 1 d and OC from the thighbone and shinbone of SD rats aged 5 d,the plane OB and OC co-culture system was established,in which the cells in each culture system did not intermix,but the culture medium communicated,and BBC-containing serum group and control group were concluded in the experiment.(2) Female SD rats aged 10 months were divided into BBC group and control group to prepare BBC-containing serum and control serum.(3) The TRAP secreted by OC was measured by the diazol method and the absorption of lacuna formed by OC was observed by microscope in OB and OC co-culture system to determine the activity of OC with the effect of BBC-containing serum.(4) The effect of BBC-containing serum on OC apoptosis in OB and OC co-culture system was examined by optical microscopes and fluorescence microscopes.RESULTS: In OB and OC co-culture system,the activity of TRAP secreted by OC of BBC-containing serum group were obviously lower than that of control group in 48 h,72 h and 96 h,the OC surviving numbers were obviously lower than that of control group and the rate of OC apoptosis was markedly higher than that of control group which showed an obvious time-efficiency relationship;the amount of the absorption of lacuna formed by OC in BBC-containing serum group was evidently lower than that of control group(P<0.01).CONCLUSION: The BBC-containing serum could significantly control the activity of OC and induce the apoptosis of OC in OB and OC co-culture system.