磷酸化南极磷虾肽调控Wnt/β-连接蛋白通路改善大鼠骨质疏松症  

Phosphorylated Peptides from Antarctic Krill Improving Osteoporosis in Ovariectomized Female Rats through Wnt/β-catenin Signaling Pathway

作　　者：王一名[1] 毛相朝[1] 曹雪[1] 王姗姗[1] 王静凤[1] Wang Yiming ,Mao Xiangzhao, Cao Xue, Wang Shanshan ,Wang Jingfeng(College of Food Science and Engineering, Ocean University of China, Qingdao 266003, Shandon)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《中国食品学报》2018年第3期8-15,共8页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家现代农业产业技术体系(CARS-48);山东省重点研发计划(2016YYSP017);国家自然科学基金(31371876)

摘　　要：目的:研究磷酸化南极磷虾肽(PP-AK P)对双侧去卵巢骨质疏松症模型大鼠的改善作用及其机制。方法:采用双侧卵巢切除法建立骨质疏松症大鼠模型,将SD大鼠随机分为5组:假手术组(生理盐水)、模型对照组(生理盐水)、阳性对照组【阿伦磷酸钠1 mg/(kg·d)】和磷酸化南极磷虾肽低、高剂量组【400,800 mg/(kg·d)】,灌胃12周后,分别检测血清骨代谢相关指标、骨密度及骨组织形态学;测定Wnt/β-连接蛋白信号通路关键基因m RNA和蛋白的表达。结果:磷酸化南极磷虾肽能显著下调去卵巢骨质疏松症大鼠血清中骨源性碱性磷酸酶(BALP)、骨钙素(OCN)和骨形态发生蛋白(BMP-2)的分泌(P<0.01),降低血清抗酒石酸酸性磷酸酶(TRAP)和I型胶原羧基端交联肽(CTX-I)水平(P<0.01),降低去卵巢大鼠骨代谢的高转换率;显著提高模型大鼠股骨骨密度(P<0.01),并能显著增加骨小梁厚度和面积,降低骨小梁间距,改善骨组织微观结构;q RT-PCR检测结果表明,磷酸化南极磷虾肽能显著降低LRP-5b、β-连接蛋白、RUNX2、OSX m RNA的表达水平;免疫印迹检测结果表明,磷酸化南极磷虾肽能显著降低LRP-5b、β-连接蛋白总蛋白、β-连接蛋白核蛋白及RUNX2蛋白的表达。结论:南极磷虾肽能显著改善双侧去卵巢大鼠骨质疏松症,其作用机制与下调Wnt/β-连接蛋白通路,降低骨质疏松症大鼠高骨转换速率有关。Objective： In this study, the effects of phosphorylated peptides from Antarctic krill（PP-AKP） in bilateral ovariectomized osteoporotic rats were assessed to study their ameliorative effects of osteoporosis and explore its mechanism. Methods： The animal model of osteoporosis was established by removing bilateral ovaries. Female SD rats were divided at random into five groups： sham-operation group, OVX group, positive control group [Alendronate Sodium, 1 mg/（kg·d）] and phosphorylated peptides from Antarctic krill groups [400, 800 mg/（kg·d）]. The rats were intragastric administrated with saline or tested substances respectively for 12 weeks. The parameters of bone metabolism, bone mineral density and histomorphology were measured at the end of the experiment. The effects of PP-AKP on Wnt/β-catenin signaling pathway were further investigated through real-time quantitative polymerase chain reaction（q RT-PCR） or Western blotting. Results： All the main parameters of bone metabolism（Bone alkaline phosphatase, osteocalein, bone morphogenetic protein-2, tartrate resistant acid phosphatase and C-terminal cross-linked telepeptide of type I collagen） reduced in PP-AKP groups, which meant phosphorylated peptides from Antarctic krill could significantly decrease bone turnover of ovaritomized rats. Bone mineral density and bone tissue microstructure werer restored in PP-AKP groups. PP-AKP decreased the transcription of LRP-5 b, β-catenin, RUNX2 and OSX. The protein expression of LRP-5 b, β-catenin and RUNX2 were also down-regulated. Conclusion： Phosphorylated peptides from Antarctic krill could significantly improve osteoporosis in bilateral ovaritomized osteoporotic rats, which is related to down-regulating Wnt/β-catenin signaling pathway, and decreasing bone turnover rate.

关 键 词：磷酸化南极磷虾肽 双侧去卵巢大鼠 高骨转换率 WNT Β-连接蛋白 

分 类 号：R580[医药卫生—内分泌]