基于Wnt信号通路探讨原花青素改善骨质疏松症的作用与分子机制  

Effect of Proanthocyanidins on Improving Osteoporosis and Its Molecular Mechanism via Wnt Signaling Pathway

作　　者：冯阳阳[1] 赵程锦[1] 周煜虎[1] 曹博[1] 段明明[1] 张亮亮 FENG Yangyang;ZHAO Chengjin;ZHOU Yuhu;CAO Bo;DUAN Mingming;ZHANG Liangliang(Department of Orthopedics and Trauma,Yanan University Affiliated Hospital,Yan’an,Shaanxi,716000,China)

机构地区：[1]延安大学附属医院创伤骨科,陕西省延安市716000

出　　处：《医学分子生物学杂志》2023年第2期103-109,共7页Journal of Medical Molecular Biology

基　　金：国家自然科学基金(No.82160431)。

摘　　要：目的基于Wnt信号通路探讨原花青素改善骨质疏松症的作用与分子机制。方法取50只SD雌性大鼠通过卵巢切除术构建骨质疏松症大鼠模型,将建模成功的大鼠随机分为模型组、阿仑膦酸钠组和原花青素低、中、高剂量组。另取10只SD大鼠记为对照组,对照组仅翻动双侧卵巢不做切除。阿仑膦酸钠组按1 mg/kg灌胃给予阿仑膦酸钠;原花青素低、中、高剂量组分别按15、25、35 mg/kg灌胃给予原花青素;对照组、模型组分别灌胃给予等量生理盐水。酶联免疫测试法(ELISA)测定各组大鼠血清氧化应激相关指标[丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)]水平和骨代谢指标[抗酒石酸酸性磷酸酶(TRAP)、Ⅰ型胶原羧基末端肽(PICP)、血清骨钙素(BGP)]含量;骨密度扫描仪扫描检测各组大鼠股骨的骨密度(BMD)值;苏木素-伊红(HE)染色观察各组大鼠骨组织形态学变化;实时荧光定量聚合酶链式反应(RT-PCR)和蛋白免疫印迹法(WB)检测骨组织Wnt、β-连环蛋白(β-catenin)、成骨标志蛋白[Runt相关转录因子2(Runx2)、Osterix]mRNA和蛋白表达。结果与对照组比较,模型组、阿仑膦酸钠组和原花青素低、中、高剂量组大鼠血清MDA、TRAP升高,SOD、GSH-Px、PICP、BGP降低(P<0.05);与模型组比较,阿仑膦酸钠组和原花青素低、中、高剂量组大鼠股骨血清MDA、TRAP降低,SOD、GSH-Px-PICP、BGP升高(P<0.05)。对照组大鼠骨小梁呈网状排列、连接紧密。模型组骨小梁数量降低、排列稀疏紊乱、连续性差。与模型组比较,阿仑膦酸钠组和原花青素低、中、高剂量组骨小梁数量及结构上均存在不同程度的改善。与对照组比较,模型组、阿仑膦酸钠组和原花青素低、中、高剂量组大鼠骨组织Wnt、β-catenin、Runx2、Osterix mRNA及蛋白表达均降低(P<0.05),与模型组比较,阿仑膦酸钠组和原花青素低、中、高剂量组大鼠骨组织Wnt、βObjective To explore the effect and molecular mechanism of proanthocyanidins on the improvement of osteoporosis based on Wnt signaling pathway.Methods The osteoporosis rat model was constructed in 50 SD female rats by ovariectomy,then they were randomly divided into model group,alendronate sodium group,proanthocyanidins low-,medium-,and high-dose groups.Another 10 SD rats were recorded as the control group,and the bilateral ovaries of rats in the control group were turned over only without resection.Rats in the alendronate sodium group were given 1 mg/kg alendronate sodium by gavage.Rats in the procyanidins low-,medium-,and highdose groups were given procyanidins by gavage at the dose of 15,25 and 35 mg/kg respectively.Rats in the control group and the model group were given the same amount of normal saline injection by gavage,once a day for 12 weeks.The levels of serum oxidative stress indexes[malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)]and bone metabolism indexes[tartrate resistant acid phosphatase(TRAP),type I procollagen carboxyl-terminal peptide(PICP),bone Gla-protein(BGP)]in rats of each group were measured by enzymelinked immunosorbent assay(ELISA).Bone density scanner was used to determine the value of femoral bone mineral density(BMD).Hematoxylin-eosin(HE)staining was used to observe the morphological changes of bone tissues in rats.Real time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting(WB)were used to detect the mRNA and protein expression levels of Wnt,β-catenin,and osteogenic marker proteins[runt related transcription factor 2(Runx2),Osterix].Results Compared with the control group,the serum MDA and TRAP levels in the model group,the alendronate sodium group and the procyanidins low-,medium-,and high-dose groups were increased,the levels of SOD,GSH-Px,PICP and BGP were decreased(P<0.05).Compared with the model group,the serum MDA and TRAP levels in the alendronate sodium group and the procyanidins low-,medium-,and high-dose groups

关 键 词：WNT信号通路 原花青素 骨质疏松症 

分 类 号：R592[医药卫生—老年医学] R681[医药卫生—内科学]