Wnt/β-catenin信号通路在郑氏补肝益肾丸治疗骨质疏松症大鼠中的作用  

Role of Wnt/β-catenin signaling pathway in treatment of osteoporosis rats by Zhengshi-Bugan Yishen Pills

作　　者：谭友莉[1] 巨少华[1] 周凌[1] 王强[1] 刘志杨 康亚兰 王明建[1] TAN You-li;JU Shao-hua;ZHOU Ling;WANG Qiang;LIU Zhi-yang;KANG Ya-lan;WANG Ming-jian(Department of Pharmacy,Sport Hospital Affiliatde To CDSU of Sichuan Province,Chengdu 610041,China)

机构地区：[1]成都体育学院附属体育医院药学部,成都610041

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2023年第6期550-558,共9页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家体育总局体育社会科学、软科学研究项目(202220135)。

摘　　要：目的探讨郑氏补肝益肾丸对骨质疏松的影响及其作用机制。方法60只SD大鼠随机分为假手术组、模型组、阳性组(阿仑膦酸钠)和郑氏补肝益肾丸低、中、高剂量组。假手术组切除卵巢附近部分脂肪组织,其余各组均采用切除双侧卵巢法建立骨质疏松症大鼠模型。术后7 d,郑氏补肝益肾丸低、中、高剂量组分别灌胃给药0.315、0.63、1.26 g/kg,阳性组灌胃给药0.5 mg/kg阿仑膦酸钠,假手术组和模型组灌胃等量生理盐水,1次/d,连续灌胃10周。micro-CT扫描股骨远端重建三维图像,扫描电镜观察股骨干骺端骨结构,HE染色观察骨组织病理损伤情况,抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)染色统计破骨细胞数目,ELISA检测血清骨钙素(osteocalcin,OC)、降钙素(calcitonin,CT)、碱性磷酸酶(alkaline phosphatase,ALP)水平,Western blot检测股骨组织Wnt3a和β-catenin蛋白表达。结果与假手术组比较,模型组骨密度(bone mineral density,BMD)、骨体积比(bone volume/tissue volume,BV/TV)、骨小梁数目(trabecular number,Tb.N)、骨小梁厚度(trabecular thickness,Tb.Th)和连接密度(connection density,Conn.D)显著降低(P<0.001),骨小梁分离度(trabecular separation,Tb.Sp)和结构模型指数(structure model index,SMI)显著升高(P<0.001);大鼠骨小梁结构不完整、变薄,骨质变得疏松,骨髓腔明显增大,腔内细胞显著变少;破骨细胞数量(P<0.01)、OC水平(P<0.001)明显增加,CT、ALP(P<0.001)水平、Wnt3a和β-catenin(P<0.01)蛋白表达显著降低。采用郑氏补肝益肾丸干预治疗后,BMD、BV/TV、Tb.N、Tb.Th和Conn.D显著增加(P<0.01或P<0.001),Tb.Sp和SMI显著降低(P<0.001);骨小梁结构有所完整,排列更有序,骨髓腔明显减少,骨质流失有所减少,破骨细胞数量明显减少(P<0.001),OC水平显著降低(P<0.05或P<0.001),CT、ALP水平(P<0.01或P<0.001)、Wnt3a(P<0.01)和β-catenin(P<0.05)蛋白表达显著增加。结论郑氏补肝益肾丸对骨�Objective To investigate the effect of Zhengshi-Bugan Yishen Pills on osteoporosis and its mechanism.Methods Sixty 3-month-old SD female rats were randomly divided into sham group,model group,positive group(alendronate),and low,medium,high-dose Zhengshi-Bugan Yishen Pills groups.In sham group,part of the adipose tissue near the ovary was removed,and in other groups,the osteoporotic rat model was established by ovariectomy.On postoperative day 7,Zhengshi-Bugan Yishen Pills were administered by gavage at the doses of 0.315,0.63,and 1.26 g/kg in low,medium and high-dose groups,respectively,while sham group and model group were given normal saline once a day for 10 weeks.The positive group was administered 0.5 mg/kg alendronate sodium by gavage.The micro-CT scan was used to observe the three-dimensional reconstruction of distal femur.Scanning electron microscope was used to observe bone structure of femoral shaft sticking end.HE staining was used to observe the pathological injury of bone tissue.Tartrate-resistant acid phosphatase(TRAP)staining was used to count the number of osteoclasts.ELISA was used to detect the levels of serum osteocalcin(OC),calcitonin(CT),and alkaline phosphatase(ALP).The protein expressions of Wnt3a andβ-catenin were detected by Western blot.Results Compared with sham group,bone mineral density(BMD),bone volume/tissue volume(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),and connection density(Conn.D),were significantly decreased(P<0.001),and trabecular separation(Tb.Sp)and structure model index(SMI)were significantly increased(P<0.001).The trabecular bone structure was incomplete,and the bone became thin.The bone became loose,and the bone marrow cavity was significantly enlarged.The cells in the cavity were significantly reduced,and the number of osteoclasts(P<0.01),OC level significantly increased(P<0.001).CT and ALP levels(P<0.001),and protein expression of Wnt3a andβ-catenin(P<0.01)were significantly decreased in model group.After Zhengshi-Bugan Yishen Pills were given as in

关 键 词：郑氏补肝益肾丸 骨质疏松症 WNT/Β-CATENIN 骨吸收 骨形成 

分 类 号：R681[医药卫生—骨科学]