Traumatic compressive stress inhibits osteoblast differentiation through long chain non-coding RNA Dancr
Traumatic compressive stress inhibits osteoblast differentiation through long chain non-coding RNA Dancr
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创伤性压缩应力通过长链非编码RNA Dancr抑制成骨细胞分化
DOI：
10.1002/jper.19-0648
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Dingming Huang
中科院分区：
文献类型：
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作者：
Qian Lu;Weizhe Xu;Linyi Liu;Xuedong Zhou;Ling Ye;Dongzhe Song;Lan Zhang;Dingming Huang
研究背景咬合创伤是牙周病加重牙周袋和牙槽骨吸收的重要局部因素。我们以前的研究发现咬合创伤通过核因子（NF）-κB信号通路抑制成骨分化。方法以MC 3 T3-E1细胞为体外研究对象，分别用单纯应激负荷、Dancr-siRNA +应激负荷、Dancr-过表达质粒+应激负荷刺激细胞，观察长链非编码分化拮抗非蛋白编码RNA（Dancr）在创伤应激对成骨细胞分化抑制中的作用。采用定量真实的实时聚合酶链反应检测Dancr、碱性磷酸酶（Alp）和Runt相关转录因子2（Runx 2）的RNA表达水平。Western印迹法检测Alp和Runx 2蛋白表达，Alp染色定性显示Alp活性。结果创伤性压应力可抑制MC 3 T3-E1细胞Alp、Runx 2和Dancr的表达，并抑制NF-κB信号通路的变化。在沉默Dancr后，应力诱导的成骨细胞分化抑制得到促进。过表达Dancr可在一定程度上缓解创伤力对成骨细胞分化的抑制作用。结论创伤性压应力可通过下调Dancr间接激活NF-κ B信号，从而抑制成骨分化。
BackgroundOcclusal trauma is an important local contributing factor aggravating periodontal pocket and alveolar bone absorption in periodontal diseases. Our previous studies have found that occlusal trauma inhibited osteogenic differentiation through nuclear factor (NF)‐κB signaling. To further investigate the underlying mechanism, the aim of this study was to explore the role of long chain non‐coding differentiation antagonizing non‐protein coding RNA (Dancr) in the inhibitory effect of traumatic stress on osteoblast differentiation.MethodsWe took the MC3T3‐E1 cells as object in vitro research and stimulated cells with simple stress load, Dancr‐siRNA + stress load, Dancr overexpression‐plasmid + stress load. Quantitative real‐time polymerase chain reaction was used to detect the RNA expression levels of Dancr, alkaline phosphatase (Alp) and Runt‐related transcription factor 2 (Runx2). The protein expressions of Alp and Runx2 were tested by Western blot and the activity of Alp was qualitatively demonstrated by Alp staining. In addition, Western blot was performed to investigate the role of Dancr in affecting NF‐κB signaling pathway.ResultsTraumatic compressive stress inhibited the expressions of Alp, Runx2, andDancr in MC3T3‐E1 cells. Stress‐induced inhibition of osteoblast differentiation was promoted after silencing Dancr. Overexpression of Dancr could alleviate the inhibitory effect of traumatic force on osteoblast differentiation to some extent. Furthermore, NF‐κB signaling was activated after silencing Dancr, and the activated effect of traumatic force on NF‐κB signaling could be alleviated through overexpression of Dancr to some extent.ConclusionTraumatic compressive stress can indirectly activate the NF‐κB signaling through downregulation of Dancr, thereby inhibiting osteogenic differentiation.