Isobavachin promotes neurogenesis of mouse embryonic stem cells <italic style="font-style: italic">in vitro</italic> by regulating miR-124

Bei WANG; Chenyan YUAN; Ruiqin YAO

doi:10.16016/j.1000-5404.201812009

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Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124

Basic Medicine | 更新时间：2026-01-09

- Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124
- Journal of Third Military Medical University Vol. 41, Issue 12, Pages: 1110-1116(2019)
- 作者机构：
  
  1.210009 南京，东南大学附属中大医院检验科
  2.221004 江苏徐州，徐州医科大学生物学教研室
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.201812009
  CLC： R322.8;R329.21;R394.2
- Received：03 December 2018，
  
  Revised：2019-03-05，
  
  Published：30 June 2019
- 稿件说明：
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Bei WANG, Chenyan YUAN, Ruiqin YAO. Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124[J]. Journal of Third Military Medical University, 2019, 41(12): 1110-1116.
DOI：

Bei WANG, Chenyan YUAN, Ruiqin YAO. Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124[J]. Journal of Third Military Medical University, 2019, 41(12): 1110-1116. DOI： 10.16016/j.1000-5404.201812009.

摘要

目的

探讨异补骨脂甲素(isobavachin，IBA)调控微小RNA-124(miR-124)对小鼠J1胚胎干细胞向神经细胞分化的影响。

方法

采用qRT-PCR和免疫荧光染色分别检测IBA干预或抑制miR-124表达的J1细胞中miR-124、八聚体转录因子4(octamer transcription factor-4，OCT4)和神经微丝H(neurofilament-H

NFH)的表达。使用Targetscan在线预测、双荧光素酶报告基因和Western blot实验验证miR-124和锌指蛋白18(zinc finger and BTB domain containing 18

ZBTB18)的靶向关系。在J1细胞中转染pcDNA-ZBTB18或miR-124 mimics、ZBTB18 shRNA，免疫荧光染色检测细胞中OCT4和NFH的表达。

结果

IBA可显著抑制J1细胞中OCT4蛋白表达，并上调miR-124和NFH表达；而抑制miR-124表达后所得结果相反。miR-124可靶向调控ZBTB18蛋白表达。在J1细胞中过表达ZBTB18，OCT4阳性表达率升高而NFH阳性表达率降低；敲减ZBTB18可在一定程度上减弱miR-124对IBA干预J1细胞OCT4和NFH表达的促进或抑制作用。

结论

IBA能够促进J1细胞中miR-124表达，而miR-124可靶向调控ZBTB18，从而调节OCT4和NFH蛋白表达，这可能是IBA促进小鼠胚胎干细胞分化为神经细胞的作用机制。

Abstract

Objective

To investigate the effect of isobavachin (IBA) on neurogenesis of mouse J1 embryonic stem cells and explore the mechanism in light of microRNA-124 (miR-124) regulation.

Methods

The expression of miR-124

octamer transcription factor-4 (OCT4) and neurofilament-H (NFH) were detected by qRT-PCR in J1 cells after treatment with IBA or transfection with miR-124 inhibitor. Targetscan online prediction

dual-luciferase assay and Western blotting were used for verifying the targeting of miR-124 to Zinc finger and BTB domain containing 18 (ZBTB18). The expression levels of OCT4 and NFH were detected with immunofluorescence assay in J1 cells transfected with pcDNA-ZBTB18 or with miR-124 mimics along with ZBTB18 shRNA.

Results

IBA significantly inhibited the expression of OCT4 and promoted the expression of miR-124 and NFH

and inhibition of miR-124 obviously enhanced OCT4 expression in J1 cells. The rsults of Targetscan online prediction

dual-luciferase assay and Western blotting indicated that miR-124 regulated the expression of ZBTB18 protein. Overexpression of ZBTB18 significantly increased the expression of OCT4 and decreased the expression of NFH in IBA-treated J1 cells; ZBTB18 knockdown in the cells attenuated the regulatory effects of miR-124 on the expression of OCT4 and NFH in IBA-treated J1 cells.

Conclusion

IBA promotes the expression of miR-124 in J1 cells

and the latter regulates the expression of OCT4 and NFH proteins by targeting ZBTB18

which can be a possible mechanism by which IBA promotes neurogenesis of mouse embryonic stem cells

in vitro

关键词

Keywords

references

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Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124

DOI：10.16016/j.1000-5404.201812009

摘要

Abstract

关键词

Keywords

references