骨细胞程序性坏死对绝经后骨质疏松症患者脆性骨折的影响  

Effects of osteocytes necroptosis on fragility fracture in patients with postmenopausal osteoporosis

作　　者：崔红旺 苏天 温鹏 黎祥涛 陈丽英 付昆[2] CUI Hong-wang;SU Tian;WEN Peng;LI Xiang-tao;CHEN Li-ying;FU Kun(Department of Emergency and Trauma Surgery,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,China;Department of Joint Surgery,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院急诊和创伤外科,海口570102 [2]海南医学院第一附属医院关节外科,海口570102

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2023年第5期458-467,共10页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：海南省重点研发计划(ZDYF2021SHFZ084)。

摘　　要：目的 明确绝经后骨质疏松(postmenopause osteoporosis, PMOP)脆性骨折患者骨细胞是否发生程序性坏死,探讨骨细胞程序性坏死对PMOP患者脆性骨折的影响,进一步完善PMOP的发病机制。方法 收集2020年1月至2021年1月于海南医学院第一附属医院PMOP股骨颈脆性骨折患者的临床病例资料和骨组织标本。采用TUNEL+受体相关蛋白激酶(receptor interaction protein kinase, RIP)3免疫荧光检测骨组织中发生程序性坏死的骨细胞数量,免疫荧光和免疫组化检测程序性坏死标志性蛋白RIP1/3、混合谱系激酶配体(mixed lineage kinase domain-like, MLKL)及凋亡标志性蛋白cleaved caspase-3的表达,并比较两组间的差异,透射电镜观察骨细胞的形态学改变,micro-CT重建股骨颈Ward’s三角骨组织微结构;采用Pearson相关性检验分析骨细胞程序性坏死率与骨组织微结构的相关性。结果 免疫荧光和免疫组化显示:PMOP股骨颈脆性骨折患者RIP1、RIP3、MLKL、cleaved caspase-3蛋白于骨细胞胞质均有表达,且TUNEL+/RIP3+(程序性坏死细胞)(19.97%±1.83%vs. 5.12%±0.29%)和TUNEL+/cleaved caspase-3+(凋亡细胞)(17.18%±1.32%vs. 3.26±0.41%)百分率较正常组明显增高(P<0.01);免疫组化结果显示:RIP1(49.38%±2.87%vs. 23.36%±1.25%)、RIP3(26.12%±4.13%vs. 6.74%±0.96%)、MLKL(29.63%±1.23%vs. 7.96%±0.42%)阳性细胞百分率较正常组明显增高(P<0.01)。透射电子显微镜发现,PMOP股骨颈脆性骨折患者骨细胞可见典型的坏死形态学特征;PMOP股骨颈脆性骨折患者Ward’s三角出现明显的骨量丢失,即骨密度[(725.63±10.17)g/cm3vs.(775.19±11.26)g/cm3]、骨小梁厚度(trabecular thickness, TB.Th)[(0.13±0.01)mm vs.(0.16±0.02)mm]、骨小梁数量(trabecular number, TB.N)[(1.35±0.04)mm-1vs.(1.52±0.07)mm-1]小于对照组(P<0.01);而骨小梁分离度(trabecular separation, TB.Sp)(0.73±0.15)mm vs.(0.61±0.21)mm大于对照组(P<0.01)。Pearson相关分析显示,骨细胞程序性坏死与骨密�Objective To explore the impact of osteocytes necroptosis in fragility fracture of patients with postmenopausal osteoporosis(PMOP),and further improve the pathogenesis of PMOP.Methods The clinical data and bone tissue samples of patients with femoral neck fragility fracture due to PMOP in the First Affiliated Hospital of Hainan Medical University from January 2020 to January 2021 were collected.TUNEL+receptor interaction protein kinase(RIP)3 immunofluorescence was used to detect the number of necroptotic osteocytes in bone tissue.The expressions of necroptotic marker proteins RIP1,RIP3,mixed lineage kinase domain-like(MLKL),and apoptotic marker protein cleaved caspase-3 were detected by immunofluorescence and immunohistochemistry,and compare the differences between the two groups.The morphological changes of osteocyte were observed by transmission electron microscope.Reconstruction of microstructure of ward s triangle bone of femur by Micro-CT.Pearson correlation test was used to analyze the correlation between the rate of osteocyte necroptosis and microstructure of bone tissue.Results The results of immunofluorescence and immunohistochemistry showed:RIP1,RIP3,MLKL,and cleaved caspase-3 proteins were expressed in the cytoplasm of osteocytes in PMOP patients with fragility fracture of femoral neck,and the percentages of TUNEL+/RIP3+positive cells(necroptotic cells)(19.97%±1.83%vs.5.12%±0.29%)and TUNEL+/cleaved caspase-3+positive cells(apoptotic cells)(17.18%±1.32%vs.3.26%±0.41%)were significantly higher than those in control group(P<0.01).The percentage of RIP1,RIP3,MLKL(49.38%±2.87%,26.12%±4.13%,29.63%±1.23%)were detected by immunohistochemistry were significantly higher than those in the normal bone mass group(23.36%±1.25%,6.74%±0.96%,7.96%±0.42%)(P<0.01).Under transmission electron microscopy,we found that osteocytes in patients with PMOP femoral neck fragility fracture showed typical morphological characteristics of necrosis.The bone loss of ward s triangle in PMOP patients with fragility fracture of

关 键 词：绝经后骨质疏松 脆性骨折 程序性坏死 骨细胞 受体相关蛋白激酶3 

分 类 号：R681[医药卫生—骨科学]