基于RANKL/RANK/OPG信号通路探讨土贝母苷甲对糖尿病性骨质疏松大鼠骨质流失的影响  

Exploration on effects of Tubeimoside Ⅰ on bone loss in diabetic osteoporosis rats based on RANKL/RANK/OPG signaling pathway

作　　者：杨晓净 邓向群 肖婷 杨梅 YANG Xiaojing;DENG Xiangqun;XIAO Ting(Department of Endocrine,Optics Valley Campus,Wuhan Third Hospital,Hubei,Wuhan 430000,China)

机构地区：[1]湖北省武汉市第三医院光谷院区内分泌科,430000

出　　处：《河北医药》2023年第2期174-179,共6页Hebei Medical Journal

基　　金：湖北省自然科学基金资助项目(编号:ZRMS2017001529)。

摘　　要：目的 探讨土贝母苷甲对糖尿病性骨质疏松(OP)大鼠骨质流失及核因子κB受体活化因子配体(RANKL)/核因子κB受体活化因子(RANK)/骨保护蛋白(OPG)信号通路的影响。方法 采用高脂高糖饮食+链脲佐菌素腹腔注射法构建大鼠糖尿病性OP模型,将60只Wistar大鼠随机分为对照组、模型组、土贝母苷甲低剂量组(1μmol/L)、土贝母苷甲高剂量组(5μmol/L)、OPG组(3 mg/kg)、土贝母苷甲高剂量+OPG组(5μmol/L土贝母苷甲+3 mg/kg OPG)。酶联免疫吸附(ELISA)法测定大鼠血清核心结合因子α1(CBF-α1)、Ⅰ型前胶原氨基端原肽(PINP)、骨钙素(OC)水平;Micro-CT测定骨密度(BMD)及骨小梁微结构参数;TRAP染色测定股骨表面破骨细胞数量(N.Oc/BS);HE染色观测大鼠骨组织病理变化;qRT-PCR法与Western blot法分别测定骨组织RANKL、RANK、OPG mRNA与蛋白水平。结果 与对照组相比,模型组大鼠血清CBF-α1、PINP、OC水平、股骨最大负荷、断裂挠度、BMD、BV/TV、Tb.N、Tb.Th、OPG mRNA与蛋白水平降低(P<0.05),Tb.Sp、N.Oc/BS、RANKL mRNA与蛋白、RANK mRNA与蛋白表达升高(P<0.05);与模型组相比,土贝母苷甲高剂量组、OPG组血清CBF-α1、PINP、OC水平、股骨最大负荷、BMD、BV/TV、Tb.N、Tb.Th、OPG mRNA与蛋白水平升高(P<0.05),Tb.Sp、N.Oc/BS、RANKL mRNA与蛋白、RANK mRNA与蛋白表达降低(P<0.05);与土贝母苷甲高剂量组相比,土贝母苷甲高剂量+OPG组大鼠上述指标均进一步升高或降低(P<0.05)。结论 土贝母苷甲可能通过抑制RANKL/RANK/OPG通路,减少糖尿病性OP大鼠破骨细胞形成,抑制骨流失。Objective To explore the effects of Tubeimoside Ⅰ(TBMS1) on the bone loss and receptor activator of nuclear factor κB ligand(RANKL)/receptor activator of nuclear factor κB(RANK)/osteoprotegerin(OPG) signaling pathway in diabetic osteoporosis(OP) rats.Methods The diabetic OP model for rats was set up with the high fat and high sugar diet + intraperitoneal injection of streptozotocin.Sixty Wistar rats were randomly assigned into the control group,model group,and low-dose TBMS1 group(1μmol/L),high-dose TBMS1 group(5μmol/L),OPG group(3mg/kg),high-dose TBMS1 + OPG group(5μmol/L TBMS1 +3mg/kg OPG).Enzyme linked immunosorbent assay(ELISA) was applied to measure the level of serum core binding factor α1(CBFα1),serum procollagen type I N-terminal propeptide(PINP) in rats,and osteocalcin(OC);micro-computed tomography(Micro-CT) was applied to measure the bone mineral density(BMD) and parameters of trabecular bone microstructure;tartrate-resistant acid phosphatase(TRAP) staining was applied to determine the osteoclast number on the femur bone surface(N.Oc/BS);HE staining was applied to observe the pathological changes of rat bone tissues;the quantitative real-time polymerase chain reaction(qRT-PCR) and Western blot were used to determine the level of RANKL,RANK,OPG mRNA and proteins in bone tissues,respectively.Results Compared with the control group,serum CBFα1,PINP,OC level,maximum femoral load,fracture deflection,BMD,Bone volume/total volume(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),OPG mRNA and protein level of rats in the model group decreased(P<0.05),and the expression of trabecular separation(Tb.Sp),N.Oc/BS,RANKL mRNA & protein,RANK mRNA & protein increased(P<0.05).Compared with the model group,the serum CBFα1,PINP,OC level,maximum femoral load,BMD,BV/TV,Tb.N,Tb.Th,OPG mRNA & protein leve of rats in the high-doseTBMS1 group and OPG group increased(P<0.05),the expression of Tb.Sp,N.Oc/BS,RANKL mRNA & protein,RANK mRNA and protein decreased(P<0.05).The above indicators in the high-dose TBMS1+

关 键 词：糖尿病性骨质疏松 核因子κB受体活化因子配体/核因子κB受体活化因子/骨保护蛋白 土贝母苷甲 

分 类 号：R587.1[医药卫生—内分泌]