金雀异黄酮酊对去卵巢骨质疏松大鼠OPG/RANKL/RANK通路的影响

Effects of Genistein Tincture on OPG/RANKL/RANK pathway in ovariectomized osteoporotic rats

目的观察金雀异黄酮酊对去卵巢骨质疏松大鼠骨密度、血清骨代谢生物标志物以及骨保护素(osteoprotegerin,OPG)、核因子κB受体活化因子(receptor activator of NF-κB,RANK)和破骨细胞分化因子(receptor activator of NF-κB,RANKL)表达的影响,探讨其抗骨质疏松的作用机制。方法将30只3月龄SD雌性大鼠随机分为假手术组、模型组和金雀异黄酮酊组,通过去卵巢法构建骨质疏松大鼠模型,给予金雀异黄酮酊外用喷涂,治疗8周后分别测定大鼠股骨骨密度,采用Elisa法检测血清骨碱性磷酸酶(ALP)、骨钙素(OC)及血清Ⅰ型胶原交联C-末端肽(S-CTX)等骨代谢生物标志物,采用Western Blot法检测OPG和RANKL蛋白表达水平。结果与假手术组比较,去卵巢组大鼠子宫质量、血清雌激素水平、股骨骨密度、血钙、血磷和OPG蛋白表达水平均降低,而体质量、ALP、OC、S-CTX和RANKL蛋白表达水平均升高,差异有统计学意义(P<0.05);采用金雀异黄酮酊治疗8周后,与模型组比较,金雀异黄酮酊组体质量、ALP、OC、S-CTX和RANKL蛋白表达水平降低,而子宫质量、血清雌激素水平、股骨骨密度、血钙、血磷和OPG蛋白表达水平均升高,差异有统计学意义(P<0.05)。结论金雀异黄酮酊可调节骨代谢,显著提高骨质疏松大鼠的骨密度,可能与调控OPG/RANKL/RANK通路蛋白表达水平有关。

Objective To investigate the effect of Genistein Tincture on bone mineral density (BMD),serum biomarkers of bone metabolism,osteoprotegerin (OPG),the receptor activator of NF-κ B (RANK) and the receptor activator of NF-κ B (RANKL) in ovariectomized rat models with osteoporosis,and to explore the mechanism. Methods Thirty 3-month-old SD rats were randomly divided into sham group,model group and Genistein Tincture group.Osteoporosis rat model was established by ovariectomy.The Genistein Tincture was given by topical spraying.After 8 weeks of treatment,the femur bone mineral density was measured respectively.Serum bone alkaline phosphatase (ALP),osteocalcin (OC),serum C-terminal telopeptide of typeⅠcollagen (S-CTX) and other biomarkers of bone metabolism were detected by Elisa.The expression of OPG and RANKL protein were detected by Western Blot. Results Compared with the sham operation group,the uterus weight,serum estrogen level,femur bone mineral density,serum calcium,serum phosphorus and OPG protein express levels in ovariectomized rats were decreased,while the body weight,ALP,OC,S-CTX,RANKL protein express levels were increased,and the differences were statistically significant ( P <0.05).After topical spraying Genistein Tincture for 8 weeks,compared with the model group,the body weight,ALP,OC,S- CTX and RANKL protein expression levels were decreased in Genistein Tincture group,while the uterus weight,serum estrogen level,femur bone mineral density,serum calcium,serum phosphorus and OPG protein express levels were increased,and the differences were statistically significant ( P <0.05). Conclusion Genistein Tincture can regulate the bone metabolism and significantly increase the bone mineral density in osteoporotic rats,which may be related to the regulation of OPG/RANKL/RANK pathway protein expression.