毕杨, 林彤远. 尼古丁调控血管平滑肌细胞内生物钟基因表达节律的研究[J]. 蚌埠医学院学报, 2020, 45(11): 1458-1461, 1466. DOI: 10.13898/j.cnki.issn.1000-2200.2020.11.002
    引用本文: 毕杨, 林彤远. 尼古丁调控血管平滑肌细胞内生物钟基因表达节律的研究[J]. 蚌埠医学院学报, 2020, 45(11): 1458-1461, 1466. DOI: 10.13898/j.cnki.issn.1000-2200.2020.11.002
    BI Yang, LIN Tong-yuan. Study on the nicotine regulating the circadian clock gene expression rhythm in vascular smooth muscle cells[J]. Journal of Bengbu Medical College, 2020, 45(11): 1458-1461, 1466. DOI: 10.13898/j.cnki.issn.1000-2200.2020.11.002
    Citation: BI Yang, LIN Tong-yuan. Study on the nicotine regulating the circadian clock gene expression rhythm in vascular smooth muscle cells[J]. Journal of Bengbu Medical College, 2020, 45(11): 1458-1461, 1466. DOI: 10.13898/j.cnki.issn.1000-2200.2020.11.002

    尼古丁调控血管平滑肌细胞内生物钟基因表达节律的研究

    Study on the nicotine regulating the circadian clock gene expression rhythm in vascular smooth muscle cells

    • 摘要:
      目的研究尼古丁对原代大鼠主动脉血管平滑肌细胞(VSMCs)内生物钟基因表达节律的影响。
      方法使用贴壁法提取原代VSMCs,免疫荧光法检测VSMCs标志物α-SMA的表达,MTT法检测细胞活力,RT-PCR法检测VSMCs内生物钟基因Bmal1、CLOCK、Per1、Cry1的表达节律。数据使用余弦法拟合,计算昼夜节律参数。
      结果尼古丁在100 nmol/L浓度刺激48 h,诱导VSMCs增殖效果最好。Bmal1和CLOCK基因的中值相对于对照组下调(P < 0.05和P < 0.01),而Per1和Cry1基因表达的中值上调,CLOCK基因表达振幅相对于对照组下调,而Per1和Cry1的振幅上调,差异均有统计学意义(P < 0.05~P < 0.01);尼古丁使VSMCs内生物钟基因表达相位后移4~7 h。
      结论尼古丁在48 h内使VSMCs内生物钟基因表达变化的同时,使VSMCs特异性增殖,产生表型分化。

       

      Abstract:
      ObjectiveTo study the effects of nicotine on the circadian clock gene expression rhythm in primary rat aortic vascular smooth muscle cells(VSMCs).
      MethodsThe primary VSMCs were extracted using adherent method.The expression level of marker α-SMA of VSMCs was detected using immunofluorescence method, the viability was detected using MTT assay, and the expression rhythm of Bmal1, CLOCK, Per1 and Cry1 genes in VSMCs cells was detected using RT-PCR.The data were fitted using the cosine method to calculate circadian parameters.
      ResultsThe 100 nmol/L of nicotine could induce the best proliferation of VSMCs for 48 hours.Compared with the control group, the median value of Bmal1 and CLOCK genes expression were down-regulated(P < 0.05 and P < 0.01), the median value of Per1 and Cry1 genes expression were up-regulated, the amplitude of Per1 and Cry1 genes expression were down-regulated, the amplitude of Per1 and Cry1 genes expression were up-regulated, and the differences of which were statistically significant(P < 0.05 to P < 0.01).
      ConclusionsNicotine can change the circadian clock gene expression in VSMCs within 48 hours, and induce specific proliferation of VSMCs, and produce phenotypic differentiation.

       

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