摘要
目的:探讨胞质羧肽酶1(CCP1)对成年小鼠骨稳态的调控作用。方法:取8周龄野生型小鼠和CCP1基因敲除小鼠,通过整体骨骼的阿利新蓝和茜素红染色、苏木精-伊红(HE)染色和Micro-CT观察对照组和敲除组小鼠大体形态和股骨骨量的变化;免疫荧光染色观察对照组和敲除组小鼠股骨内成骨活动的变化;免疫荧光染色观察对照组和敲除组小鼠股骨内破骨活动的变化,实时荧光定量PCR检测破骨细胞相关基因表达变化。结果:与对照组小鼠比较,CCP1缺失后小鼠骨发育迟缓,股骨骨量降低(P<0.01),股骨骨小梁厚度(Tb.Th)和骨皮质厚度(Cort.Th)降低(P<0.05),股骨内Runt相关转录因子2(RUNX2)阳性骨祖细胞的数量减少(P<0.05)、牙本质基质蛋白1(DMP1)基因表达降低(P<0.05),股骨内破骨细胞数量显著增加(P<0.01),抗酒石酸酸性磷酸酶(TRAP)和组织蛋白酶K(CTSK)表达增加(P<0.05),核因子κB受体活化子配体(RANKL)表达增加(P<0.01),骨保护素(OPG)基因表达降低(P<0.05),RANKL/OPG比值增加(P<0.01)。结论:CCP1缺失引起小鼠骨发育迟缓和骨量降低,成骨活动减弱而破骨活动增强,CCP1可能发挥促进骨形成的作用。
Objective:To investigate the regulatory role of cytosolic carboxypeptidase 1(CCP1)in bone homeostasis of adult mice.Methods:Eight-week-old wild-type(WT)and CCP1 knockout(KO)mice were used.Whole-mount skeletal staining with alcian blue and alizarin red was performed to assess gross morphology.Hematoxylin-eosin(HE)staining and Micro-CT were employed to evaluate bone microstructure and bone mass.Immunofluorescence staining was conducted to analyze osteogenic activity in the femurs.Immunofluorescence staining was performed to examine osteoclastic activity.Quantitative real-time PCR was used to measure the expression levels of osteoclast-related genes.Results:CCP1 deficiency causes delayed bone development in mice;Loss of CCP1 leads to reduced bone mass in mouse femurs(P<0.01);CCP1 KO mice exhibit lower trabecular thickness(Tb.Th),and cortical thickness(Cort.Th)in the femur(P<0.05);CCP1-deficient femurs exhibit decreased number of Runt-related transcription factor 2-positive(RUNX2+)osteoprogenitor cells and lower dentin matrix protein 1(DMP1)expression(P<0.05).Concurrently,CCP1 knockout significantly increases the number of osteoclasts(P<0.01),along with increased expression of cathepsin K(CTSK)and tartrate-resistant acid phosphatase(TRAP)(P<0.05).Furthermore,femurs from CCP1 KO mice exhibit increased receptor activator of nuclear factorκB ligand(RANKL)expression level(P<0.01),reduced osteoprotegerin(OPG)expression level(P<0.05),and a higher RANKL/OPG ratio(P<0.01).Conclusions:CCP1 deficiency leads to delayed bone development,reduced bone mass,and impaired osteogenic activity,while promoting osteoclastic resorption.These findings suggest that CCP1 plays a critical role in promoting bone formation.
作者
潘灿灿
孙瑶
PAN Cancan;SUN Yao(Shanghai Engineering Research Center of Tooth Restoration and Regeneration&Tongji Research Institute of Stomatology&Department of Oral Implantology,Shanghai Tongji Stomatological Hospital and Dental School,Tongji University,Shanghai 200072,China)
出处
《口腔生物医学》
2025年第4期206-211,217,共7页
Oral Biomedicine
基金
上海市教育委员会科研创新计划项目(2023ZKZD29)。
关键词
胞质羧肽酶1
骨稳态
成骨
破骨
cytosolic carboxypeptidase 1
bone homeostasis
osteogenesis
osteoclastogenesis
