仙灵骨葆胶囊对去卵巢骨折大鼠骨折端骨微结构的影响及OPG/RANKL信号调控机制  

Effect of Xianling Gubao Capsules on microstructure of fractured bone in ovariectomized rats and regulation mechanism of OPG/RANKL signaling

作　　者：任世元 张琦豪 闫晓哲 REN Shi-yuan;ZHANG Qi-hao;YAN Xiao-zhe(Zhengzhou Municipal Orthopedics Hospital,Zhengzhou 450052,China)

机构地区：[1]郑州市骨科医院,河南郑州450052

出　　处：《中成药》2021年第1期67-72,共6页Chinese Traditional Patent Medicine

基　　金：2015年度郑州市第三批科技计划资助项目(153PKJGG084)。

摘　　要：目的研究仙灵骨葆胶囊对去卵巢大鼠骨折端骨微结构的影响及骨保护素(OPG)/核因子κB受体活化因子配基(RANKL)信号调控机制。方法从48只Sprague-Dawley雌鼠中随机挑出40只切除双侧卵巢,恢复性饲养6周后建立股骨骨折内固定模型,建模大鼠随机分为模型组,阳性对照组及仙灵骨葆胶囊高、中、低剂量组,余下8只作为正常组。模型组和正常组大鼠灌胃给予2 mL生理盐水,每天1次;阳性对照组大鼠灌胃给予0.018 mg/mL戊酸雌二醇(10 mL/kg),每天1次,连续6 d后停药1 d;仙灵骨葆胶囊高、中、低剂量组分别将100、50、25 mg/kg药物溶于生理盐水中灌胃,每天1次。连续治疗10周后,酶联免疫吸附实验(ELISA)检测大鼠血清骨碱性磷酸酶(BALP)、骨钙素(OC)、Ⅰ型胶原交联c-末端肽(s-CTX)水平;双能X线扫描仪对大鼠骨折愈合时间情况进行观察,并测定大鼠骨折处骨密度;苏木精-伊红(HE)染色观察大鼠骨折端骨微结构愈合情况;实时定量聚合酶链式反应(qRT-PCR)检测大鼠骨折处股骨组织中OPG、RANKL mRNA表达;蛋白免疫印记法(WB)检测大鼠骨折处骨组织中OPG、RANKL蛋白表达。结果仙灵骨葆胶囊干预组骨折愈合时间低于模型组(P<0.05);仙灵骨葆胶囊干预组大鼠血清BALP、OC水平高于模型组(P<0.05),血清s-CTX水平低于模型组(P<0.05);仙灵骨葆胶囊干预组大鼠股骨骨折处骨密度高于模型组(P<0.05),骨小梁结构多于模型组(P<0.05)。仙灵骨葆胶囊干预组OPG mRNA、蛋白表达比模型组高(P<0.05),RANKL mRNA和蛋白表达量比模型组低(P<0.05)。结论仙灵骨葆胶囊能减少去卵巢骨折大鼠骨折愈合时间,增加骨折处骨密度,升高血清BALP和OC水平,降低s-CTX水平,减轻股骨骨折端微结构断裂缺损状况,促进去卵巢骨折大鼠断骨愈合,其作用机制可能与上调OPG mRNA和蛋白表达,下调RANKL mRNA和蛋白表达有关。AIM To study the effect of Xianling Gubao Capsules on microstructure of fractured bone in ovariectomized rats and regulation mechanism of osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)signaling.METHODS Forty rats were randomly selected from 48 female Sprague-Dawley rats for 6 weeks of recovery feeding,and establishment of femoral fracture models with internal fixation.The rats were randomly divided into model group,positive control group,and Xianling Gubao Capsule groups for use of high dose,medium dose,and low dose agent administration.The remaining 8 rats were recorded as negative control group.Both model group and negative control group were given 2 mL of normal saline by gavage once daily.The positive control group was dosed 0.018 mg/mL of estradiol valerate at 10 mL/kg once daily for 6 consecutive days and discontinued for 1 day.Rats in Xianling Gubao Capsule groups were intervened with the capsules dissolved in 2mL of normal saline by gavage at concentrations of 100 mg/kg,50 mg/kg and 25 mg/kg respectively once daily.After 10 weeks of continuous treatment,the rats were subjected to the detection of serum bone alkaline phosphatase(BALP),osteocalcin(OC)and s-C-terminal telopeptide(s-CTX)levels by ELISA;the observation of the fracture healing time and measurement of bone mineral density at fracture site by dual-energy X-ray scanner;observation of the microstructure healing of fracture tip by hematoxylin eosin staining(HE);detection of OPG,RANKL mRNA expressions at the fracture site in femur bone by quantitative real-time polymerase chain reaction(qRTPCR);determination of levels of OPG and RANKL protein expression by western blot(WB).RESULTS In contrast to the model group,Xianling Gubao Capsule groups shared shorter fracture healing time(P<0.05),higher levels of serum BALP and OC(P<0.05),and lower serum s-CTX level(P<0.05),higher bone density of the femoral fractures(P<0.05),more trabecular structure(P<0.05),higher levels of OPG mRNA and protein expression(P<0.05),and lower levels of RANKL mRNA

关 键 词：仙灵骨葆胶囊 骨微结构 骨保护素 核因子κB受体活化因子配基 

分 类 号：R285.5[医药卫生—中药学]