基于Wnt信号通路探讨补肾益气活血中药复方对去卵巢大鼠骨质疏松症的作用机制

Study on the mechanism of the effect of tonifying kidney, invigorating qi, and activating blood circulation Chinese herbal compound on osteoporosis in ovariectomized rats based on Wnt signaling pathway

目的观察去卵巢大鼠骨质疏松症模型骨、肾组织中Wnt3a信号蛋白的变化,探究补肾益气活血中药复方对骨质疏松症作用的分子生物学机制。方法将雌性大鼠75只随机分为5组:正常组、模型组、福善美组、补肾益气活血方低剂量组、补肾益气活血方高剂量组,后4组摘除大鼠双侧卵巢法复制骨质疏松症模型,连续给药12周后以双能X线检测骨密度,ELISA法检测血清碱性磷酸酶(alkaline phosphatase,ALP)、抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)以及骨、肾组织中Wnt3a蛋白表达。结果与正常组比较,模型组骨密度明显降低,血清ALP明显减少,血清TRAP明显升高,骨及肾组织中Wnt3a蛋白表达明显减少;与模型组相比较,各治疗组骨密度、血清ALP及骨、肾组织Wnt3a蛋白均有不同程度的升高。结论骨质疏松症的发生机制与Wnt3a蛋白变化有关;补肾益气活血中药复方的疗效作用机制是通过调节和改善Wnt3a蛋白的表达,激活Wnt3a/β-catenin信号传导通路,促成骨分化,抑制破骨细胞,起到防治骨质疏松症的作用。

Objective To observe the changes of Wnt3 a signal protein in the bone and kidney of ovariectomized rats, and to explore the molecular biological mechanism of the effect of compound prescription of tonifying kidney, invigorating qi, and activating blood circulation on osteoporosis. Methods Seventy-five female rats were randomly divided into five groups: normal group, model group, Fosamax group, low-and high-dose of tonifying kidney, invigorating qi and activating blood circulation compound group. The rats in the latter four groups were ovariectomized to establish osteoporosis model. After 12 weeks of continuous administration of the drugs, bone mineral density was measured with dual energy X-ray and serum levels of ALP and TRAP were detected with ELISA method. Wnt3 a protein expression in bone and kidney was examined. Results Compared with the normal group, the bone mineral density, serum ALP and TRAP, and Wnt3 a protein expression in the bone and kidney significantly decreased in the model group. Compared with the model group, the bone mineral density, serum ALP, and Wnt3 a protein in the bone and kidney significantly increased in each treatment group. Conclusion The mechanism of the development of osteoporosis is related to the change of Wnt3 a protein. The therapeutic mechanism of tonifying kidney, invigorating qi, and activating blood circulation herbal compound is to regulate and improve the expression of Wnt3 a protein, activate Wnt3 a/?-catenin signal transduction pathway, promote osteogenic differentiation, and inhibit osteoclasts, Results ing in prevention and treatment of osteoporosis.