益肾蠲痹汤含药血清对大鼠成骨细胞和破骨细胞增殖能力的影响

目的 通过研究益肾蠲痹汤的含药血清对大鼠成骨细胞、破骨细胞增殖的影响，评价益肾蠲痹法对强直性脊柱炎骨代谢异常的治疗作用。 方法 SD正常大鼠(SPF级)经灌胃给药益肾蠲痹汤，收集大鼠含药血清。分离大鼠成骨细胞和破骨细胞，通过Van kossa染色和碱性磷酸酶染色鉴定成骨细胞，TRAP染色、甲苯胺蓝染色鉴定破骨细胞。将体外培养的成骨细胞和破骨细胞，分别分为6组:空白对照组，益肾蠲痹汤大、中、小剂量含药血清组，双氯芬酸对照组和阿仑膦酸盐对照组。使用CCK-8法测定成骨细胞、破骨细胞的生长曲线，收集并测定成骨细胞培养液上清碱性磷酸酶活性。 结果 含有益肾蠲痹汤药物的血清加入体外培养成骨细胞后，成骨细胞的生长曲线呈明显上升趋势，加药后细胞培养上清中碱性磷酸酶浓度增加，提示成骨细胞分泌碱性磷酸酶活性增强。生长曲线上升趋势及分泌碱性磷酸酶活性随含药血清浓度增加而作用增强。体外培养破骨细胞加入益肾蠲痹汤含药血清后，细胞增殖曲线呈下降趋势，曲线下降趋势亦随含药血清浓度升高而幅度增大。 结论 益肾蠲痹汤具有促进成骨细胞增殖及提高成骨细胞活性，同时，对破骨细胞增殖有抑制作用，作用强度随药物浓度增加而增强。因此，预期益肾蠲痹汤对强直性脊柱炎骨代谢异常具有治疗作用。 

Effect of serum containing Yishen Juanbi Decoction on proliferation of rat osteoblasts and osteoclasts

Objective The explore the preventive effects of Yishen Juanbi Decoction on bone metabolism disorder associated with ankylosing spondylitisby observing the effect of Yishen Juanbi Decoction on proliferation of rat osteoblasts and osteoclasts. Methods Yishen Juanbi decoction was prepared firstly,and the serum containing Yishen Juanbi decoction was prepared following intragastricgavage to rats.The osteoblasts and osteoclastswere isolated and differentiated by stainings of Van kossa,ALP,TRAP and toluidine blue staining.The cells were divided into placebo-treated group,low,medium and large doses of serum containing Yishen Juanbi decoction groups,Diclofenac Sodium control group and Alendronate Sodiumcontrol group.The Cell Counting Kit 8(CCK8) test was conducted to assessthe activities of osteoblast and osteoclast cells.The alkaline phosphatase activityin the cell culture supernatant was tested. Results The serum containing Yishen Juanbi decoction significantly promoted the proliferation and alkaline phosphatase secretion of rat osteoblast in vitro,and increased with the levels of serum.On the other hand,the decoction serum significantly inhibited the proliferation of rat osteoclast in vitro,positively with the contents of serum. Conclusions Yishen Juanbi Decoction can promote the proliferation of osteoblast and improve the activity of osteoblast,andsimultaneously inhibit the proliferation of osteoclast,the effects increased with the doses of decoction.The experiment suggests that Yishen Juanbi Decoction can improve the bone metabolism disorder associated with ankylosing spondylitis.