| 王 媛,王钧左,陈 励,严溢泉,赵星成,孙喜庆.时钟基因Bmal1促进血管平滑肌细胞增殖[J].现代生物医学进展英文版,2022,(16):3010-3013. |
| 时钟基因Bmal1促进血管平滑肌细胞增殖 |
| Clock Gene Bmal1 Promotes Proliferation of Vascular Smooth Muscle Cell |
| Received:December 11, 2021 Revised:December 30, 2021 |
| DOI：10.13241/j.cnki.pmb.2022.16.003 |
| 中文关键词: Bmal1 血管平滑肌细胞 增殖 昼夜节律 |
| 英文关键词: Bmal1 Vascular smooth muscle cell Proliferation Circadian rhythm |
| 基金项目:陕西省自然科学基础研究计划项目（2020JM-317）；国家自然科学基金项目（81803098） |
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| 中文摘要: |
| 摘要 目的：观察时钟基因Bmal1的过表达对血管平滑肌细胞增殖的影响，进一步探讨生物节律对于血管发育的具体影响。方法：采用包装GV341-Bmal1载体的慢病毒转染的方法构建大鼠胸主动脉平滑肌细胞（A7R5）稳定转染Bmal1的细胞系，实时定量PCR和细胞爬片Bmal1的免疫荧光染色的方法判断所构建细胞系是否稳定过表达Bmal1，细胞爬片Ki67的免疫荧光染色的方法观察时钟基因Bmal1的过表达对血管平滑肌细胞增殖的影响。结果：实时定量PCR结果显示稳定转染Bmal1组细胞Bmal1的表达是对照组的11.2倍（P<0.01）；细胞爬片的免疫荧光染色结果显示稳定转染Bmal1组细胞BMAL1的表达明显升高（P<0.05），且稳定转染Bmal1组Ki67阳性细胞比例明显升高（P<0.05）。结论：通过慢病毒转染的方法成功构建了血管平滑肌细胞稳定转染Bmal1的细胞系，细胞片Ki67的免疫荧光染色结果显示Bmal1的过表达促进了血管平滑肌细胞的增殖。 |
| 英文摘要: |
| ABSTRACT Objective: To observe the effect of overexpression of clock gene Bmal1 on the proliferation of vascular smooth muscle cells and investigate potential role mechanism of biological rhythms on vascular developmen through experiments. Methods: The method of lentiviral transfection packaging GV341-Bmal1 vector was used to construct rat thoracic aortic smooth muscle cells (A7R5) stably transfected with Bmal1 cell line. Real-time fluorescent quantitative PCR(RT-PCR) and immunofluorescence staining of Bmal1 on the cells grown on coverslips were used to determine whether the stable transfected cell line overexpressed Bmal1 stably. Immunofluorescence staining of Ki67 on the cells grown on coverslips was used to observe the effect of overexpression of clock gene Bmal1 on the proliferation of vascular smooth muscle cells. Results: The results of Real-time fluorescent quantitative PCR showed that the expression of Bmal1 in cells stably transfected with Bmal1 was 11.2 times higher compared to control group(P<0.01). The results of immunofluorescence staining of the cells grown on coverslips showed that the expression of BMAL1 was significantly increased in stable transfected cell lines(P<0.05). Furthermore, compared to the control group, the proportion of Ki67 positive cells was also increased significantly in vascular smooth muscle cells stably transfected with Bmal1(P<0.05). Conclusion: We have established a vascular smooth muscle cell line with Bmal1 overexpressed stably by Lentiviral transfection method. Through observing immunofluorescence staining of Ki67 on the cells grown on coverslips, we have found that overexpression of Bmal1 promoted the proliferation of vascular smooth muscle cells. |
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