云南白药用于骨质疏松治疗的动物疗效研究

The anti-osteoporotic effect of the Chinese traditional medicine Yunnan Baiyao on osteoprotegerin knockout mice

目的探讨云南白药对骨质疏松是否具有治疗效果。方法采用护骨素(osteoprotegerin,OPG)基因敲除小鼠为骨质疏松模型,将7月龄OPG基因剔除小鼠(OPG^(-/-))分为三组(每组5只),分别给予云南白药治疗、密盖息治疗和生理盐水治疗。作为对照,将同样背景的野生型小鼠(OPGWT)分为2组(每组3只),分别给予云南白药和生理盐水治疗。药物干预6个月后,分别检测小鼠血清ALP、ACP水平,行离体骨密度测定、骨生物力学测试,并采用皮下注射盐酸四环素和钙黄绿素进行骨表面的双荧光标记,进行骨形态计量学参数测定,将数据进行统计学分析。结果 OPG^(-/-)云南白药治疗组和OPG(-/-)密盖息治疗组在ALP浓度水平上较生理盐水对照组均有显著性的升高(P<0.05)。OPG(-/-)云南白药治疗组的骨矿化沉积率参数较生理盐水对照组有显著上升(P<0.05)。云南白药治疗组在骨强度、骨形态计量学静态参数上有改善,但和生理盐水组对比,尚无显著差异。结论研究发现云南白药可促使OPG基因敲除模型小鼠血清碱性磷酸酶活性显著升高,骨矿化沉积率显著上升,提示该药可刺激成骨细胞的功能,具有一定的改善骨质疏松的作用。

Objective To study the anti-osteoporotic effect of Chinese traditional medicine Yunnan Baiyao and provide basic data for further study on its mechanism and application in clinic. Methods Seven-month-old male osteoprotegerin（OPG） knockout mice（ OPG -/- ） were randomly divided into 3 groups （5 mice per group）, and treated with Yunnan Baiyao, Miacalcic and normal saline respectively. Background and age matched wild type mice（OPG＋/＋） were divided into 2 groups（3 mice per group）, which were treated with Yunnan Baiyao and normal saline respectively as control. Parameters including serum alkaline phosphatase （ALP）, acid phosphatase （ACP）, bone mineral density （BMD）, biomechanical testing and bone histomorphometry were measured after 6 months of treatment. Results were analyzed statistically. Results Compared with the normal saline treated OPG-/- group, the serum ALP level in Yunnan Baiyao OPG -/- treatment group and OPG -/- Miacalcic treatment group were significantly increased（ P 〈 0.05 ） and mineral apposition rate（MAR） in Yunnan Baiyao OPG -/- treatment group was also significantly increased （ P 〈 0.05 ） after 6 months treatment. The results of biomechanical testing and bone histomorphometry showed the parameters were getting better in OPG-/- Yunnan Baiyao treatment group but had no significant differences with control group. Conclusion Yunnan Baiyao can increase the serum ALP and MAR in OPG knockout mice which indicates that it can stimulate the function of osteoblast. This result provides basic data to fin/her study the antiosteoporotic mechnisim of Yunnan Baiyao and its potential clinical application in osteoporosis treatment.