摘要
目的 探讨人脂质运载蛋白2（Lipocalin-2,LCN2）基因在骨质疏松症中的作用及机制.方法 反转录-聚合酶链反应（RT-PCR）法检测并比较骨质疏松患者（n=18）及非骨质疏松患者（n=18）血液标本Lipocalin-2 mRNA表达量;下调小鼠骨髓间充质干细胞（BMSCs） Lipocalin-2基因,碱性磷酸酶（ALP）染色茜素红染色检测细胞成骨分化能力,Westem blot检测细胞成骨分化相关基因表达;建立骨质疏松小鼠模型,下调骨质疏松小鼠Lipocalin-2基因,micro-CT检测小鼠胫骨上段骨小梁体积,检测股骨抗弯的力学性能.结果 骨质疏松患者组（1.693±0.359） Lipocalin-2的mRNA相对表达量高于非骨质疏松患者组（1.108±0.234）,P=0.019.而且在骨质疏松患者中,有骨质疏松性骨折病史患者的Lipocalin-2的mRNA相对表达量（n=10,2.172±0.624）高于无骨折病史的患者组（n=8,1.473±0.336）.Lipocalin-2下调组小鼠BMSCs成骨分化基因Runt相关基因2（Runχ2）、骨钙蛋白（OCN）的蛋白水平及mRNA相对表达量分别高于对照组.Lipocalin-2下调组[（812.78 ±25.36） N/mm]小鼠股骨抗弯强度高于对照组[（744.36±31.54） N/mm],LCN2-小干扰RNA（siRNA）组[（19.69±3.69） μm2]小鼠胫骨上段骨小梁面积大于对照组[（14.77±4.25）μm2].结论 骨质疏松患者血液Lipocalin-2表达上升,下调Lipocalin-2基因后干细胞成骨分化能力增强,下调Lipocalin-2基因能改善小鼠骨质疏松程度,其机制与Runχ2及OCN相关.
Objective To investigate the role and mechanism of Lipocalin-2 gene in osteoporosis.Methods Reverse transcriptase-polymerase chain reaction （RT-PCR） assay and compared patients with osteoporosis （n =18） and non osteoporosis patients （n =18） expression of Lipocalin-2 blood samples by mRNA;mouse bone marrow mesenchymal stem cells with Lipocalin-2 gene,alkaline phosphatase （ALP） staining and alizarin red staining to detect cell osteogenic differentiation of Western cells,blotting detection of bone differentiation related gene expression of bone;a mouse model of osteoporosis,osteoporosis by mouse Lipocalin-2 gene,micro-CT detection of mouse tibia trabecular bone volume,the mechanical properties of the femur bending test.Results The relative expression of mRNA in patients with osteoporosis （1.693 ± 0.359） Lipocalin-2 was higher than that of non osteoporotic group （1.108 ± 0.234）,P =0.019.Moreover,the relative expression of Lipocalin-2 mRNA in osteoporotic patients with osteoporotic fracture history （n =10,2.172 ± 0.624） is higher than that in patients without fracture history （n =8,1.473 ± 0.336）.The protein level of Runχ2 and osteocalcin （OCN） and the relative expression of mRNA in the bone marrow mesenchymal stem cells （BMSCs） osteogenic differentiation gene of Lipocalin-2 mice were higher than those in the control group.In the Lipocalin-2 group [（812.78 ±25.36） N/mm],the femoral flexural strength was higher than that in the control group [（744.36 ±31.54） N/mm],while in the LCN2-small interfering RNA （siRNA） group [（19.69 ± 3.69） μm2],the trabecular area in the proximal tibia was larger than that in the control group [（14.77 ± 4.25） μm2].Conclusion The expression of Lipocalin-2 is increased in patients with osteoporosis,and the ability of osteogenic differentiation of stem cells is enhanced after downregulation of Lipocalin-2 gene.Down regulation of Lipocalin-2 gene can improve the degree of osteoporosis in mice,and its mechanism is related to Ru
作者
谢远龙
孙文超
谢江涛
Xie Yuanlong;Sun Wenchao;Xie Jiangtao(Department of Orthopaedics, Zhongnan Hospital of Wuhan University, Wuhan 430071, China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第5期881-884,共4页
Chinese Journal of Experimental Surgery
关键词
骨质疏松症
骨髓间充质干细胞
人脂质运载蛋白2
成骨分化
Osteoporosis
Bone marrow mesenchymal stem cells
Human lipid carrier protein 2
Osteogenic differentiation
