基于OPG/RANK/RANKL信号通路研究补肾强督方对大鼠去卵巢骨质疏松的作用机制  

Study on Bushen Qiangdu formula based on OPG/RANK/RANKL signaling pathway mechanism of action on ovariectomized osteoporosis rats

作　　者：王宇秀 杨永生 石嘉颖 吴昊[3] WANG Yuxiu;YANG Yongsheng;SHI Jiaying;WU Hao(Inner Mongolia Fourth Hospital,Hohhot 010051,China)

机构地区：[1]内蒙古自治区第四医院,内蒙古呼和浩特010051 [2]呼和浩特市蒙医中医医院,内蒙古呼和浩特010050 [3]内蒙古医科大学,内蒙古呼和浩特010107

出　　处：《陕西中医》2024年第10期1330-1335,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：内蒙古自治区科技计划项目(2021GG0186);第五批全国中医临床优秀人才研修项目[国中医药人教函(2022)1号]。

摘　　要：目的:观察补肾强督方干预去卵巢所致骨质疏松(OP)大鼠对OPG/RANK/RANKL信号通路的影响,以探索补肾强督方防治绝经后骨质疏松(PMOP)的机制。方法:选取40只SPF级雌性SD大鼠适应性喂养1周后称重并按照体重大小从低到高依次编号,然后利用随机数字表法分为假手术组(6只)和手术组(34只),手术组中有4只因为手术原因死亡,剩余30只,手术组切除双侧卵巢构建OP大鼠模型后分为模型对照组、阿仑膦酸钠组、补肾强督方高剂量组、补肾强督方中剂量组、补肾强督方低剂量组各6只。假手术组仅切除等重量卵巢旁脂肪组织。药物干预10周后取大鼠左右股骨及腹主动脉采血制备血清,采用酶联免疫吸附法(ELISA)检测血清中核因子κB受体活化因子(RANK)、核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)、肿瘤坏死因子受体相关因子6(TRAF6)、骨钙素(OCN)水平情况,分别采用蛋白质免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应法(RT-PCR)检测骨组织OPG、RANK、RANKL蛋白及mRNA的表达,并进行统计学处理。结果:补肾强督方可以促进血清中OPG含量以及股骨中OPG蛋白及mRNA的表达,抑制血清中RANK、RANKL、TRAF6、OCN含量和股骨中RANK、RANKL蛋白及mRNA的表达。结论:补肾强督方可能通过调控OPG/RANKL/RANK信号通路以及下游TRAF6因子的表达抑制破骨细胞增殖、刺激成骨细胞的表达,动态调节骨代谢的平衡。Objective:To observe the effect of Bushen Qiangdu decoction on OPG/RANKL/RANK signaling pathway in ovariectomized osteoporosis(OP)rats,and to explore the mechanism of Bushen Qiangdu decoction in the prevention and treatment of postmenopausal osteoporosis(PMOP).Methods:A total of 40 SPF female SD rats were selected after adaptive feeding for 1 week and weighed and numbered according to the weight from low to high.Then they were divided into sham operation group(6 rats)and operation group(34 rats)by random number method.After the bilateral ovaries of the operation group were removed to construct the OP rat model,they were divided into model control group,alendronate sodium group,high dose group of Bushen Qiangdu decoction,middle dose group of Bushen Qiangdu decoction and low dose group of Bushen Qiangdu decoction.The sham operation group only removed the same weight of paraovarian adipose tissue.After 10 weeks of drug intervention,blood samples were collected from the left and right femurs and abdominal aortas of rats to prepare serum.The levels of receptor activator of nuclear factor-κB(RANK),receptor activator of nuclear factor-κB ligand(RANKL),osteoprotegerin(OPG),tumor necrosis factor receptor-associated factor 6(TRAF6)and osteocalcin(OCN)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The expression of OPG,RANK and RANKL protein and mRNA in bone tissue were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction(RT-PCR),and statistical analysis was performed.Results:Bushen Qiangdu decoction can promote the expression of OPG in serum and OPG protein and mRNA in femur,inhibit the expression of RANK,RANKL,TRAF6,OCN in serum and RANK,RANKL protein and mRNA in femur.Conclusion:Bushen Qiangdu decoction may inhibit the proliferation of osteoclasts and stimulate the expression of osteoblasts by regulating the RANK/RANKL/OPG signaling pathway and the expression of downstream TRAF6 factors,and dynamically regulate the balance of bone metabolism.

关 键 词：骨质疏松 补肾强督方 骨保护素 核因子ΚB受体活化因子配体 核因子ΚB受体活化因子 肿瘤坏死因子受体相关因子6 

分 类 号：R681[医药卫生—骨科学]