补肾壮筋汤调控OPG/RANKL/RANK信号通路抗骨质疏松的作用  

Role of Bushen Zhuangjin Decoction Against Osteoporosis by Regulating OPG/RANKL/RANK Signaling Pathway

作　　者：周世博 俞兴[2] 仲文庆 李文浩 赵赫[2] 熊洋[2] ZHOU Shibo;YU Xing;ZHONG Wenqing;LI Wenhao;ZHAO He;XIONG Yang(Beijing University of Chinese Medicine,Beijing 100029,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东直门医院,北京100700

出　　处：《中国实验方剂学杂志》2024年第16期44-51,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(82104901,82305273);北京中医药大学新教师启动基金项目(2023-BUCMXJKY010)。

摘　　要：目的:观察补肾壮筋汤(BSZJ)对去卵巢大鼠骨代谢、骨密度(BMD)、骨小梁微结构及对骨保护素/核转录因子-κB受体活化因子配体/核转录因子-κB受体活化因子(OPG/RANKL/RANK)信号通路的影响。探索补肾壮筋汤抗骨质疏松作用机制。方法:32只雌性SD大鼠随机分为假手术组(SHAM)、卵巢去势模型组(OVX)、阿仑膦酸钠组(ALN,7.35 g·kg^(-1))和补肾壮筋汤组(BSZJ,3.2 g·kg^(-1))。卵巢切除12周后给予对应药物灌胃,共12周。12周后采集血清和骨标本。酶联免疫吸附测定法(ELISA)检测血清Ⅰ型前胶原氨基端原肽(PINP)和Ⅰ型胶原交联C-末端肽(CTX-Ⅰ),三点弯曲实验测试大鼠胫骨生物力学,微型计算机断层扫描(Micro-CT)分析腰椎BMD及骨小梁微结构。包括骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)、骨小梁分离度(Tb.Sp)及结构模型指数(SMI)。苏木素-伊红(HE)及马松(Masson)染色分析腰椎骨小梁及骨胶原变化。实时荧光定量聚合酶链式反应(Real-time PCR)测定腰椎OPG、RANKL、RANK mRNA表达。蛋白免疫印迹法(Western blot)测定腰椎OPG、RANKL、RANK蛋白的表达。结果:与SHAM组比较,OVX组血清PINP及CTX-Ⅰ明显增加(P<0.05);与OVX组比较,ALN组及BSZJ组血清CTX-Ⅰ明显降低(P<0.05)。与SHAM组比较,OVX组胫骨生物力学载荷(Fracture Load)明显降低(P<0.05);与OVX组比较,ALN组和BSZJ组生物力学载荷增加。与SHAM组比较,OVX组腰椎BMD、BV/TV、Tb.Th及Tb.N明显降低,Tb.Sp及SMI明显增加(P<0.05);与OVX组比较,BSZJ组腰椎BMD、BV/TV及Tb.N明显增加,Tb.Sp及SMI明显降低(P<0.05)。与SHAM组比较,OVX组骨小梁结构紊乱,骨小梁之间的距离增加;胶原纤维变细,连续性欠佳;与OVX组比较,BSZJ组腰椎骨小梁结构相对完整,骨小梁之间的距离变小,胶原纤维增粗。与SHAM组比较,OVX组腰椎OPG mRNA及蛋白表达明显降低,RANKL、RANK mRNA及蛋白表达明显增加(P<0.05);与OVX组比较,BSZJ组OPG mRNA�Objective:To observe the effects of Bushen Zhuangjin decoction(BSZJ)on bone metabolism,bone mineral density(BMD),bone trabecular microstructure,and osteoprotegerin/receptor activator of nuclear factor kappa B ligand/receptor activator of nuclear factor kappa B(OPG/RANKL/RANK)signaling pathway in ovariectomized rats and explore the action of mechanism of BSZJ against osteoporosis.Method:Thirty-two female SD rats were randomly divided into a sham-operated group(SHAM),an ovariectomized group(OVX),an alendronate group(ALN,7.35 g·kg^(-1)),and a BSZJ group(3.2 g·kg^(-1)).Corresponding drugs were given by gavage 12 weeks after ovarian removal,and the process lasted for 12 weeks.Serum and bone specimens were collected after 12 weeks.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum procollagen Ⅰ N-terminal propeptide(PINP)and C-telopeptide of type Ⅰ collagen(CTX-Ⅰ).Biomechanics of rat tibia was tested by a three-point bending test.BMD of lumbar vertebrae and bone trabecular microstructure were analyzed by micro-computed tomography(Micro-CT),including bone volume/tissue volume(BV/TV),trabecular thickness(Tb.Th),trabecular number(Tb.N),trabecular separation/spacing(Tb.Sp),and structure model index(SMI).Hematoxylin-eosin(HE)and Masson staining were used to analyze trabecular and collagen changes in lumbar vertebrae.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to determine the mRNA expression of OPG,RANKL,and RANK in lumbar vertebrae.Protein expression of OPG,RANKL,and RANK in lumbar vertebrae was determined by Western blot.Result:Serum PINP and CTX-Ⅰ were significantly increased in the OVX group compared with the SHAM group(P<0.05).Serum CTX-Ⅰ was significantly reduced in the ALN and BSZJ groups compared with the OVX group(P<0.05).Tibial fracture load was significantly lower in the OVX group compared with the SHAM group(P<0.05).Fracture Load was increased in the ALN and BSZJ groups compared with the OVX group.Compared with those in the SHAM group,BMD,BV/TV,T

关 键 词：骨质疏松 补肾壮筋汤 卵巢去势大鼠 骨保护素/核转录因子-κB受体活化因子配体/核转录因子-κB受体活化因子(OPG/RANKL/RANK)信号通路 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33