益骨汤对去势大鼠骨密度及经典BMP2信号通路的影响  

Effect of Yigu decoction on classical BMP2 signaling pathway in bone tissue of ovariectomized rats

作　　者：林晓芳 方芳[2] 李桂锦[3] 姚新苗[3] LIN Xiaofang;FANG Fang;LI Guijin;YAO Xinmiao(Department of Orthopedics,Taizhou Hospital of Zhejiang Province,Linhai 317000,Zhejiang,China;Laboratory of Acupuncture Neurobiology,the Third Clinical School of Medicine,Zhejiang Chinese Medicine University,Hangzhou 310053,Zhejiang,China;Department of Orthopaedics,the Third Affiliated Hospital of Zhejiang Chinese Medicine University,Hangzhou 310053,Zhejiang,China)

机构地区：[1]浙江省台州医院骨科,浙江临海317000 [2]浙江中医药大学第三临床医学院针灸神经生物学实验室,浙江杭州310053 [3]浙江中医药大学附属第三医院骨科,浙江杭州3100531

出　　处：《中国现代医生》2024年第6期73-78,83,共7页China Modern Doctor

基　　金：浙江省自然科学基金项目(LZ15H270001);浙江省中医药科技计划项目(2021ZQ048)。

摘　　要：目的观察益骨汤对去势大鼠骨组织中经典骨形态发生蛋白(bone morphogenetic protein,BMP)2信号通路的影响,旨在探讨益骨汤防治骨质疏松症(osteoporosis,OP)的作用机制。方法将44只12周龄雌性Wistar大鼠随机分为四组,每组11只。正常组不进行任何处理,模型组、戊酸雌二醇组和益骨汤组大鼠进行去势手术以模拟骨质疏松。造模成功后,分别以10ml/kg双蒸水灌胃(正常组、模型组)、0.36mg/kg戊酸雌二醇水溶液灌胃及10ml/kg益骨汤灌胃,1次/d。12周后,双能X射线骨密度仪测定股骨骨密度(bone mineral density,BMD),实时聚合酶链反应(polymerase chain reaction,PCR)检测BMP2经典信号通路BMP2、Smad1、Runx2、OSX基因的表达,免疫组织化学法、Western blot法检测BMP2经典信号通路BMP2、Smad1、Runx2、OSX蛋白的表达量。结果①双能X射线骨密度仪测定股骨骨密度,结果显示,与模型组比较,益骨汤组大鼠的BMD升高(P<0.01)。②实时荧光定量PCR检测,与模型组比较,益骨汤组大鼠的BMP2、Smad1、Runx2、Noggin基因表达量升高(P<0.05)。③免疫组织化学法检测:与模型组比较,益骨汤组大鼠的BMP2、Smad1、Runx2、OSX蛋白表达量升高(P<0.05)。④Western blot法检测:与模型组比较,益骨汤组大鼠的Smad1、Runx2、OSX蛋白含量升高(P<0.05)。结论益骨汤可能通过激活BMP2经典信号通路及抑制Noggin的高表达,促进骨形成和防治骨质疏松。Objective To observe the impact of Yigu decoction on the classic bone morphogenetic protein(BMP)2 signaling pathway in rat bone organization,aim to explore the mechanism of Yigu decoction in preventing and treating osteoporosis(OP).Methods A total of 4412-week-old female Wistar rats were randomly divided into four groups,with 11 rats in each group.The normal group did not receive any treatment,while the model group,estradiol valerate group and Yigu decoction group were castrated to simulate osteoporosis.After successful modeling,the rats were gavaged with 10ml/kg double steamed water(normal group,model group),0.36mg/kg estradiol valerate aqueous solution,and 10ml/kg Yigu decoction,once a day.After 12 weeks,bone mineral density(BMD)of the femur was measured by dual-energy X-ray absorptiometry,real-time quantitative PCR was used to detect the expression levels of BMP2,Smad1,Runx2,and OSX genes in the BMP2 classical signaling pathway.Immunohistochemistry and Western blot were used to detect the expression levels of BMP2,Smad1,Runx2,and OSX proteins in the BMP2 classical signaling pathway.Results①The bone mineral density(BMD)of the femur measured by dual-energy X-ray absorptiometry showed that compared with the model group,the BMD of the Yigu decoction group was significantly increased(P<0.01).②Using real-time quantitative PCR,it was found that compared with the model group,the expression levels of BMP2,Smad1,Runx2,and Noggin genes in the Yigu decoction group were significantly increased(P<0.05).③Immunohistochemistry showed that the protein expression levels of BMP2,Smad1,Runx2 and OSX in Yigu decoction group were increased than those in model group(P<0.05).④Western blot showed that the protein expression of Smad1,Runx2 and OSX in Yigu decoction group were higher than those in model group(P<0.05).Conclusion Yigu decoction may promote bone formation and prevent and treat osteoporosis by activating the classical BMP2 signaling pathway and inhibiting the high expression of Noggin.

关 键 词：益骨汤 去势大鼠 骨质疏松 BMP2经典信号通路 NOGGIN 

分 类 号：R33[医药卫生—人体生理学]