摘要
目的:探讨冬凌草甲素(Ori)调节Hippo/Yes相关蛋白(YAP)信号通路对骨质疏松(OP)大鼠成骨细胞凋亡的影响.方法:随机选取10只大鼠为假手术组仅分离但不切除卵巢,其余大鼠通过切除双侧卵巢建立OP模型,将造模成功的大鼠随机分为OP组、不同剂量Ori(5mg/kg Ori、10mg/kg Ori、20mg/kg Ori)组、20mg/kg Ori+Hippo/YAP 激活剂-Verteporfin(10mg/kg)组.干预后,收集腹主动脉血液,ELISA检测血清中骨钙素(BGP)、碱性磷酸酶(ALP)水平;分离股骨组织,双能X射线骨密度仪检测股骨矿含量、骨密度;HE检测股骨组织形态学变化;TUNEL检测成骨细胞凋亡变化;Western blot检测YAP、转录共激活因子PDZ结合基序(TAZ)蛋白表达.结果:假手术组大鼠股骨结构变化不明显,与假手术组相比,OP组股骨组织形态发生明 显改变,BGP、ALP水平、股骨组织矿含量、骨 密度、YAP、TAZ蛋白表达显著降低(P＜0.05),成骨细胞凋亡率显著增加(P＜0.05);与OP组相比,5mg/kg Ori组、10mg/kg Ori组、20mg/kg Ori组股骨组织形态得到改善,BGP、ALP水平、股骨组织矿含量、骨密度、YAP、TAZ蛋白表达显著增加(P＜0.05),成骨细胞凋亡率显著降低(P＜0.05),不同剂量Ori间具有统计差异(P＜0.05);与20mg/kg Ori组相比,20mg/kg Ori+V erteporfin组股骨组织形态变化加重,BGP、ALP水平、股骨组织矿含量、骨密度、YAP、TAZ蛋白表达显著降低(P＜0.05),成骨细胞凋亡率显著增加(P＜0.05).结论:Ori调节Hippo/YAP信号通路减少OP大鼠成骨细胞凋亡,减轻OP大鼠症状.
Abstract
Objective:To investigate the effect of oridonin(Ori)on osteoblast apoptosis in osteoporosis(OP)rats by regulating the Hippo/Yes-associated protein(YAP)signaling pathway.Methods:Ten rats were randomly selected as the sham surgery group,with only ovaries separated but not removed.The remai-ning rats were used to establish an OP model by removing both ovaries.The successfully modeled rats were randomly separated into the OP group,different doses of Ori(5 mg/kg Ori,10 mg/kg Ori,20 mg/kg Ori)groups,and 20 mg/kg Ori+Hippo/YAP activator-Vesteporfin(10 mg/kg)group.After the intervention,abdominal aortic blood was collected,and ELISA was applied to detect serum levels of osteocalcin(BGP)and alkaline phosphatase(ALP).The femoral tissue was separated,and a dual-energy X-ray bone density in-strument was applied to detect femoral mineral content and bone density.HE was applied to detect morpholog-ical changes in femoral tissue.TUNEL was applied to detect changes in osteoblast apoptosis.Western blot was applied to detect the expression of YAP and transcription co-activator PDZ binding motif(TAZ)proteins.Results:There was no great change in the femoral structure of the rats in the sham surgery group.Compared with the sham surgery group,there was a great change in the morphology of the femoral tissue in the OP group,the levels of BGP and ALP,femoral tissue mineral content,bone density,YAP,and TAZ protein ex-pression were greatly reduced(P＜0.05),the apoptosis rate of osteoblasts was greatly increased(P＜0.05).Compared with the OP group,the femoral tissue morphology was improved in the 5 mg/kg Ori group,10 mg/kg Ori group,and 20 mg/kg Ori group,the levels of BGP and ALP,femoral tissue mineral content,bone density,YAP,and TAZ protein expression were greatly increased(P＜0.05),the apoptosis rate of osteoblasts was greatly decreased(P＜0.05),there were statistical differences between different doses of Ori(P＜0.05).Compared with the 20 mg/kg Ori group,the morphological changes of the femoral tissue in the 20 mg/kg Ori+Verteporfin group were aggravated,the levels of BGP and ALP,femoral tissue mineral content,bone densi-ty,YAP,and TAZ protein expression were greatly reduced(P＜0.05),the apoptosis rate of osteoblasts was greatly increased(P＜0.05).Conclusion:Ori reduces osteoblast apoptosis and alleviates symptoms in OP rats by regulating the Hippo/YAP signaling pathway.
维普
万方数据