糖尿病骨质疏松症模型小鼠脂肪干细胞的成骨能力  

Osteogenic ability of adipose stem cells in diabetic osteoporosis mice

作　　者：高俞锦 彭双麟 马治超 陆诗 曹花月 王浪 肖金刚 Gao Yujin;Peng Shuanglin;Ma Zhichao;Lu Shi;Cao Huayue;Wang Lang;Xiao Jingang(School of Stomatology of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Department of Oral and Maxillofacial Surgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Orofacial Reconstruction and Regeneration Laboratory of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学口腔医学院,四川省泸州市646000 [2]西南医科大学附属医院口腔颌面外科,四川省泸州市646000 [3]西南医科大学口颌面修复重建和再生实验室,四川省泸州市646000

出　　处：《中国组织工程研究》2022年第7期999-1004,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81870746),项目负责人:肖金刚;泸州市人民政府-西南医科大学科技战略合作项目(2020LZXNYDZ09),项目负责人:肖金刚。

摘　　要：背景:长链非编码RNA(long noncoding RNA,LncRNA)-DANCR可调节包括骨代谢的多个生物学过程。课题组基因芯片结果显示糖尿病骨质疏松小鼠脂肪组织中LncRNA-DANCR显著高表达。目的:探究LncRNA-DANCR通过Wnt/β-catenin信号通路对糖尿病骨质疏松小鼠脂肪干细胞成骨能力的影响。方法:分离培养糖尿病骨质疏松小鼠和正常对照小鼠脂肪干细胞,成骨诱导3 d后,采用qRT-PCR、Western blot检测LncRNA-DANCR、Wnt信号分子β-catenin以及成骨转录因子(RUNX2、OPN)的表达。设计LncRNA-DANCR特异性siRNA转染糖尿病骨质疏松小鼠脂肪干细胞,成骨诱导3 d后,采用qRT-PCR、Western blot和碱性磷酸酶染色检测Wnt信号分子β-catenin、成骨转录因子(RUNX2、OPN)的表达以及成骨能力改变。结果与结论:①成骨诱导3 d后,糖尿病骨质疏松小鼠脂肪干细胞的LncRNA-DANCR表达水平显著高于正常对照小鼠脂肪干细胞,Wnt信号分子β-catenin和成骨转录因子RUNX2、OPN的表达显著低于正常对照小鼠脂肪干细胞;②siRNA转染糖尿病骨质疏松小鼠脂肪干细胞成骨诱导3 d后,β-catenin以及RUNX2、OPN显著高表达,成骨能力增强;③结果表明,LncRNA-DANCR通过抑制Wnt/β-catenin信号通路降低糖尿病骨质疏松小鼠脂肪干细胞的成骨能力,沉默LncRNA-DANCR可恢复其成骨能力。BACKGROUND:Long noncoding RNA(LncRNA)-DANCR can regulate multiple biological processes,including bone metabolism.Our gene microarray results showed that LncRNA-DANCR was significantly overexpressed in diabetic osteoporosis mice.OBJECTIVE:To investigate the effect of LncRNA-DANCR on the osteogenic ability of adipose stem cells in diabetic osteoporosis through Wnt/β-catenin signaling pathway.METHODS:Diabetic osteoporosis adipose-derived stem cells and control adipose-derived stem cells were isolated and cultured.After 3 days of osteogenic induction,real-time fluorescence quantitative PCR and western blot assay were used to examine the expression of LncRNA-DANCR,Wnt signaling molecules β-catenin and osteogenic transcription factors(RUNX2 and OPN).Diabetic osteoporosis adipose-derived stem cells transfected with lncRNA-DANCR-specific siRNA were designed.After 3 days of osteogenic induction,real-time fluorescence quantitative PCR,western blot assay and alkaline phosphatase staining were used to examine the expression of Wnt signaling molecules β-catenin,osteogenic transcription factors RUNX2 and OPN and osteogenic ability changes.RESULTS AND CONCLUSION:(1)After 3 days of osteogenic induction,the expression level of LncRNA-DANCR in diabetic osteoporosis adipose-derived stem cells group was significantly higher than that in control group.The expression levels of Wnt signaling moleculeβ-catenin and osteogenic transcription factors RUNX2 and OPN were significantly lower than those in control adipose-derived stem cells.(2)After 3 days of osteogenic induction,β-catenin and RUNX2 and OPN were highly expressed in the siRNA silent group,and the ability of bone formation was enhanced.(3)The results confirmed that LncRNA-DANCR decreased the osteogenic capacity of diabetic osteoporosis adipose-derived stem cells by inhibiting the Wnt/β-catenin signaling pathway.Silencing of LncRNA-DANCR can restore the reduced osteogenic capacity.

关 键 词：长链非编码RNA WNT信号通路 糖尿病 骨质疏松 骨向分化 骨修复 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]