浏览全部资源
扫码关注微信
异补骨脂甲素调控miR-124对小鼠胚胎干细胞向神经细胞分化的影响
Isobavachin promotes neurogenesis of mouse embryonic stem cells
in vitro by regulating miR-124- 第三军医大学学报 2019年41卷第12期 页码：1110-1116
- 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：
[姚瑞芹, E-mail：1970912076@qq.com YAO Ruiqin, E-mail: 1970912076@qq.com
- 基金信息：
DOI：10.16016/j.1000-5404.201812009
中图分类号： R322.8;R329.21;R394.2收稿：2018-12-03，
修回：2019-03-05，
纸质出版：2019-06-30
- 稿件说明：
移动端阅览
王贝, 袁晨燕, 姚瑞芹. 异补骨脂甲素调控miR-124对小鼠胚胎干细胞向神经细胞分化的影响[J]. 第三军医大学学报, 2019,41(12):1110-1116.
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.王贝, 袁晨燕, 姚瑞芹. 异补骨脂甲素调控miR-124对小鼠胚胎干细胞向神经细胞分化的影响[J]. 第三军医大学学报, 2019,41(12):1110-1116. DOI： 10.16016/j.1000-5404.201812009.
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
2
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
2
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
2
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
2
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
2
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
2
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
2
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
2
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
YANG P H, CHEN X, KAUSHAL S, et al. High glucose suppresses embryonic stem cell differentiation into cardiomyocytes[J]. Stem Cell Res Ther, 2016, 7: 187. DOI: 10.1186/s13287-016-0446-5.
HU J, YANG Z Y, WANG J B, et al. Zinc chloride transiently maintains mouse embryonic stem cell pluripotency by activating stat3 signaling[J]. PLoS ONE, 2016, 11(2): e0148994. DOI: 10.1371/journal.pone.0148994.
ANTONOV S A, MANUILOVA E S, DOLOTOV O V, et al. Effect of nerve growth factor on neural differentiation of mouse embryonic stem cells[J]. Bull Exp Biol Med, 2017, 162(5): 679-683. DOI: 10.1007/s10517-017-3686-1.
GUIDOLIN D, TORTORELLA C, MARCOLI M, et al. Neuroglobin, a factor playing for nerve cell survival[J]. Int J Mol Sci, 2016, 17(11): E1817. DOI: 10.3390/ijms17111817.
LIU Z J, JIANG Y Y, LI X Y, et al. Embryonic stem cell-derived peripheral auditory neurons form neural connections with mouse central auditory neurons in vitro via the α2δ1 receptor[J ] . Stem Cell Rep, 2018, 11(1): 157-170. DOI: 10.1016/j.stemcr.2018.05.006.
HUANG M L, MICHALAK A L, FISHER C J, et al. Small molecule antagonist of cell surface glycosaminoglycans restricts mouse embryonic stem cells in a pluripotent state[J]. Stem Cells, 2018, 36(1): 45-54. DOI: 10.1002/stem.2714.
WANG D Y, HU Y Z, KONG S S, et al. Promoting effects of isobavachin on neurogenesis of mouse embryonic stem cells were associated with protein prenylation[J]. Acta Pharmacol Sin, 2011, 32(4): 425-432. DOI: 10.1038/aps.2011.5.
WANG X, ZHAO Y N, XIAO Z F, et al. Alternative translation of OCT4 by an internal ribosome entry site and its novel function in stress response[J]. Stem Cells, 2009, 27(6): 1265-1275. DOI: 10.1002/stem.58.
CHIN T K, HARDING S E, EAGLES P A. Characterization of two proteolytically derived soluble polypeptides from the neurofilament triplet components NFM and NFH[J]. Biochem J, 1989, 264(1): 53-60.
李建宁, 任维, 刘一辉.外周神经系统中神经微丝蛋白研究现状[J].山东医药, 2012, 52(27): 88-90. DOI: 10.3969/j.issn.1002-266X.2012.27.035.
LI J N, REN W, LIU Y H. Current status of research on neurofilament proteins in peripheral nervous system [J]. Shandong Med J, 2012, 52(27):88-90. DOI: 10.3969/j.issn.1002-266X.2012.27.035.
CARRO M S, LIM W K, ALVAREZ M J, et al. The transcriptional network for mesenchymal transformation of brain tumours[J]. Nature, 2010, 463(7279): 318-325. DOI: 10.1038/nature08712.
TATARD V M, XIANG C M, BIEGEL J A, et al. ZNF238 is expressed in postmitotic brain cells and inhibits brain tumor growth[J]. Cancer Res, 2010, 70(3): 1236-1246. DOI: 10.1158/0008-5472.CAN-09-2249.
XIANG C, BAUBET V, PAL S, et al. RP58/ZNF238 directly modulates proneurogenic gene levels and is required for neuronal differentiation and brain expansion[J]. Cell Death Differ, 2012, 19(4): 692-702. DOI: 10.1038/cdd.2011.144.
GULYAEVA L F, KUSHLINSKIY N E. Regulatory mechanisms of microRNA expression[J]. J Transl Med, 2016, 14: 143. DOI: 10.1186/s12967-016-0893-x.
YANG H, ZHANG L L, AN J, et al. MicroRNA-mediated reprogramming of somatic cells into neural stem cells or neurons[J]. Mol Neurobiol, 2017, 54(2): 1587-1600. DOI: 10.1007/s12035-016-0115-9.
GONZÁLEZ-GIRALDO Y, CAMARGO A, LÓPEZ-LEÓN S, et al. A functional SNP in MIR124-1, a brain expressed miRNA gene, is associated with aggressiveness in a Colombian sample[J]. Eur Psychiatry, 2015, 30(4): 499-503. DOI: 10.1016/j.eurpsy.2015.03.002.
杨俊娜, 何丽, 徐陶, 等. miR-124在神经系统发育、损伤及修复中作用的研究进展[J].山东医药, 2018, 58(20): 100-103. DOI:10.3969/j.issn.1002-266X.2018.20.034.
YANG J N, HE L, XU T, et al. Update of miR-124 in the development, injury and repair of nervous system [J]. Shandong Med J, 2018, 58(20): 100-103. DOI: 10.3969/j.issn.1002-266X.2018.20.034.
WANG C M, WEI Z J, JIANG G H, et al. Neuroprotective mechanisms of miR-124 activating PI 3 K/Akt signaling pathway in ischemic stroke[J ] . Exp Ther Med, 2017, 13(6): 3315-3318. DOI: 10.3892/etm.2017.4424.
JIANG D, DU J T, ZHANG X M, et al. MiR-124 promotes the neuronal differentiation of mouse inner ear neural stem cells[J]. Int J Mol Med, 2016, 38(5): 1367-1376. DOI: 10.3892/ijmm.2016.2751.
WANG Y, WANG D S, GUO D W. MiR-124 promote neurogenic transdifferentiation of adipose derived mesenchymal stromal cells partly through RhoA/ROCK1, but not ROCK2 signaling pathway[J]. PLoS ONE, 2016, 11(1): e0146646. DOI: 10.1371/journal.pone.0146646.
RIZZINO A, WUEBBEN E L. Sox2/Oct4: A delicately balanced partnership in pluripotent stem cells and embryogenesis[J]. Biochim Biophys Acta, 2016, 1859(6): 780-791. DOI: 10.1016/j.bbagrm.2016.03.006.
LI D W, SHEN D C, TAI H F, et al. Neurofilaments in CSF as diagnostic biomarkers in motor neuron disease: a meta-analysis[J]. Front Aging Neurosci, 2016, 8: 290. DOI: 10.3389/fnagi.2016.00290.
王伟英, 李海明, 戴艺民, 等.植物锌指蛋白的功能研究进展[J].中国园艺文摘, 2016, 32(7): 3-5, 12. DOI: 10.3969/j.issn.1672-0873.2016.07.002.
WANG W Y, LI H M, DAI Y M, et al. Advances on the function of plant zinc finger protein[J]. Chin Hort Abstr, 2016, 32(7): 3-5, 12. DOI: 10.3969/j.issn.1672-0873.2016.07.002.
ZHU C, CHEN G, ZHAO Y, et al. Regulation of the development and function of B cells by ZBTB transcription factors[J]. Front Immunol, 2018, 9: 580. DOI:10.3389/fimmu.2018.00580.
MATHEW R, SEILER M P, SCANLON S T, et al. BTB-ZFfactors recruit the E3 ligase cullin 3 to regulate lymphoid effector programs[J]. Nature, 2012, 491(7425): 618-621. DOI: 10.1038/nature11548.
MAEDA T. Regulation of hematopoietic development by ZBTB transcription factors[J]. Int J Hematol, 2016, 104(3): 310-323. DOI: 10.1007/s12185-016-2035-x.
赵倩倩, 班博, 邵倩.锌指蛋白ZBTB38调控生长发育相关研究进展[J].中国儿童保健杂志, 2018, 26(1): 44-47.
ZHAO Q Q, BAN B, SHAO Q. Research advances on zinc finger protein ZBTB38 regulating growth and development[J]. Chin J Child Health Care, 2018, 26(1): 44-47.
刘淦, 陈李斌佶, 谢志芳, 等.锌指蛋白ZBTB20生物学功能的研究进展[J].第二军医大学学报, 2015, 36(5): 525-529. DOI: 10.3724/SP.J.1008.2015.00525.
LIU G, CHEN L, XIE Z F, et al. Biological functions of zinc finger protein ZBTB20: recent advance[J]. Acad J Second Mil Med Univ, 2015, 36(5): 525-529. DOI: 10.3724/SP.J.1008.2015.00525.
COHEN J S, SRIVASTAVA S, FARWELL HAGMAN K D, et al. Further evidence that de novo missense and truncating variants in ZBTB18 cause intellectual disability with variable features[J]. Clin Genet, 2017, 91(5): 697-707. DOI: 10.1111/cge.12861.
VAN DER SCHOOT V, DE MUNNIK S, VENSELAAR H, et al. Toward clinical and molecular understanding of pathogenic variants in the ZBTB18 gene[J]. Mol Genet Genomic Med, 2018, 6(3): 393-400. DOI: 10.1002/mgg3.387.
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
