产前补充牛磺酸调节Rho家族因子活性促进生长受限新生大鼠神经干细胞增殖的研究

李翔文, 李芳, 刘敬, 王燕, 付薇

中国当代儿科杂志 ›› 2016, Vol. 18 ›› Issue (11) : 1158-1165.

PDF(2121 KB)
HTML
PDF(2121 KB)
HTML
中国当代儿科杂志 ›› 2016, Vol. 18 ›› Issue (11) : 1158-1165. DOI: 10.7499/j.issn.1008-8830.2016.11.021
论著·实验研究

产前补充牛磺酸调节Rho家族因子活性促进生长受限新生大鼠神经干细胞增殖的研究

  • 李翔文1,2, 李芳1, 刘敬1,2, 王燕2, 付薇2
作者信息 +

Effect of antepartum taurine supplementation in regulating the activity of Rho family factors and promoting the proliferation of neural stem cells in neonatal rats with fetal growth restriction

  • LI Xiang-Wen1,2, LI Fang1, LIU Jing1,2, WANG Yan2, FU Wei2
Author information +
文章历史 +

摘要

目的 探讨产前补充牛磺酸通过调节Rho家族因子活性促进生长受限 (FGR)新生大鼠神经干细胞增殖的可能性,为产前补充牛磺酸促进生长受限患儿脑发育提供实验依据。方法 将24只Sprague-Dawley孕鼠随机分为对照组、FGR组、牛磺酸组 (n=8),通过全程饥饿法建立FGR模型。采用逆转录-聚合酶链反应法、免疫组化及免疫印迹法检测各组新生鼠脑组织神经干细胞特异性胞内标记物脂肪酸结合蛋白7 (FABP7)、Rho相关螺旋卷曲蛋白激酶2 (ROCK2)、ras同源基因家族成员A (RhoA)、Ras相关的C3肉毒素底物 (rac)表达水平。结果 FGR组FABP7阳性细胞OD值、FABP7mRNA及蛋白的表达低于对照组,牛磺酸组FABP7阳性细胞OD值、FABP7mRNA及蛋白的表达高于FGR组 (P < 0.05)。FGR组中RhoA、ROCK2mRNA的表达高于对照组,牛磺酸组中RhoA、ROCK2mRNA表达高于对照组但低于FGR组 (P < 0.05);FGR组中racmRNA的表达低于对照组,牛磺酸组中racmRNA表达高于FGR组及对照组 (P < 0.05);FGR组中RhoA、ROCK2蛋白表达水平高于对照组,牛磺酸组中RhoA、ROCK2蛋白表达水平低于FGR组 (P < 0.05)。结论 产前补充牛磺酸可促进FGR新生大鼠神经干细胞增殖,其机制可能与调控Rho家族因子的活性有关。

Abstract

Objective To study the possible effect of antepartum taurine supplementation in regulating the activity of Rho family factors and promoting the proliferation of neural stem cells in neonatal rats with fetal growth restriction (FGR), and to provide a basis for antepartum taurine supplementation to promote brain development in children with FGR. Methods A total of 24 pregnant Sprague-Dawley rats were randomly divided into three groups:control, FGR, and taurine (n=8 each). A rat model of FGR was established by food restriction throughout pregnancy. RT-PCR, immunohistochemistry, and Western blot were used to measure the expression of the specific intracellular markers for neural stem cells fatty acid binding protein 7 (FABP7), Rho-associated coiled-coil containing protein kinase 2 (ROCK2), ras homolog gene family, member A (RhoA), and Ras-related C3 botulinum toxin substrate (Rac). Results The FGR group had significantly lower OD value of FABP7-positive cells and mRNA and protein expression of FABP7 than the control group, and the taurine group had significantly higher OD value of FABP7-positive cells and mRNA and protein expression of FABP7 than the FGR group (P < 0.05). The FGR group had significantly higher mRNA expression of RhoA and ROCK2 than the control group. The taurine group had significantly higher mRNA expression of RhoA and ROCK2 than the control group and significantly lower expression than the FGR group (P < 0.05). The FGR group had significantly lower mRNA expression of Rac than the control group. The taurine group had significantly higher mRNA expression of Rac than the FGR and control groups (P < 0.05). The FGR group had significantly higher protein expression of RhoA and ROCK2 than the control group. The taurine group had significantly lower protein expression of RhoA and ROCK2 than the FGR group (P < 0.05). Conclusions Antepartum taurine supplementation can promote the proliferation of neural stem cells in rats with FGR, and its mechanism may be related to the regulation of the activity of Rho family factors.

关键词

牛磺酸 / 胎儿生长受限 / 神经干细胞 / Rho家族因子 / 大鼠

Key words

Taurine / Fetal growth restriction / Neural stem cell / Rho family factor / Rats

引用本文

导出引用
李翔文, 李芳, 刘敬, 王燕, 付薇. 产前补充牛磺酸调节Rho家族因子活性促进生长受限新生大鼠神经干细胞增殖的研究[J]. 中国当代儿科杂志. 2016, 18(11): 1158-1165 https://doi.org/10.7499/j.issn.1008-8830.2016.11.021
LI Xiang-Wen, LI Fang, LIU Jing, WANG Yan, FU Wei. Effect of antepartum taurine supplementation in regulating the activity of Rho family factors and promoting the proliferation of neural stem cells in neonatal rats with fetal growth restriction[J]. Chinese Journal of Contemporary Pediatrics. 2016, 18(11): 1158-1165 https://doi.org/10.7499/j.issn.1008-8830.2016.11.021

参考文献

[1] 王华伟,刘敬.胎儿生长受限及其对脑发育的不良影响[J].中国实用儿科杂志,2015,30(10):796-799.
[2] 王晓凤,刘敬.宫内生长受限对患儿远期神经行为及心理发育的影响[J].中华全科医师杂志,2012,11(3):202-204.
[3] Baschat AA.Neurodevelopment after fetal growth restriction[J].Fetal Diagn Ther,2014,36(2):136-142.
[4] Fung C,Ke X,Brown AS,et al.Uteroplacental insufficiency alters rat hippocampal cellular phenotype in conjunction with ErbB receptor expression[J].Pediatr Res,2012,72(1):2-9.
[5] 王晓凤,滕慧云,刘敬,等.孕鼠补充牛磺酸减少胎儿生长受限胎鼠脑细胞凋亡[J].中华围产医学杂志,2013,16(3):510-513.
[6] 陈慧,李坚,刘敬,等.产前补充牛磺酸促进宫内生长受限胎鼠脑细胞增殖[J].中华围产医学杂志,2011,14(10):612-617.
[7] 王晓凤,刘颖,刘敬,等.孕鼠补充牛磺酸对宫内生长受限胎鼠脑组织Ras同源基因-Rho相关卷曲螺旋蛋白激酶通路关键信号分子表达的影响[J].中华实用儿科临床杂志,2013,28(8):312-315.
[8] Jansson N,Pettersson J,Haafiz A,et al.Down-regulation of placental transport of amino acids precedes the development of intrauterine growth restriction in rats fed a low protein diet[J].J Physiol,2006,576(Pt 3):935-946.
[9] Menendez-Castro C,Fahlbusch F,Cordasic N,et al.Early and late postnatal myocardial and vascular changes in a protein restriction rat model of intrauterine growth restriction[J].PLoS One,2011,6(5):e20369.
[10] van Batenburg-Eddes T,de Groot L,Steegers EA,et al.Fetal programming of infant neuromotor development:the generation R study[J].Pediatr Res,2010,67(2):132-137.
[11] Baschat AA.Fetal growth restriction-from observation to intervention[J].J Perinat Med,2010,38(3):239-246.
[12] Baschat AA.The fetal circulation and essential organs-a new twist to an old tale[J].Ultrasound Obstet Gynecol,2006,27(4):349-354.
[13] Baschat AA.Neurodevelopment following fetal growth restriction and its relationship with antepartum parameters of placental dysfunction[J].Ultrasound Obstet Gynecol,2011,37(5):501-514.
[14] Liu RZ,Mita R,Beaulieu M,et al.Fatty acid binding proteins in brain development and disease[J].Int J Dev Biol,2010,54(8-9):1229-1239.
[15] Pasantes-Morales H,Ramos-Mandujano G,Hernández-Benítez R.Taurine enhances proliferation and promotes neuronal specification of murine and human neural stem/progenitor cells[J].Adv Exp Med Biol,2015,803:457-472.
[16] Stankiewicz TR,Linseman DA.Rho family GTPases:key players in neuronal development,neuronal survival,and neurodegeneration[J].Front Cell Neurosci,2014,8:314.
[17] Numano F,Inoue A,Enomoto M,et al.Critical involvement of Rho GTPase activity in the efficient transplantation of neural stem cells into the injured spinal cord[J].Mol Brain,2009,2:37.
[18] Katayama K,Melendez J,Baumann JM,et al.Loss of RhoA in neural progenitor cells causes the disruption of adherens junctions and hyperproliferation[J].Proc Natl Acad Sci U S A,2011,108(18):7607-7612.
[19] 刘颖,刘敬.Rho-ROCK信号通路研究进展[J].中国儿童保健杂志,2012,20(9):822-825.
[20] Koyanagi M,Takahashi J,Arakawa Y,et al.Inhibition of the Rho/ROCK pathway reduces apoptosis during transplantation of embryonic stem cell-derived neural precursors[J].J Neurosci Res,2008,86(2):270-280.
[21] Zhang L,Valdez JM,Zhang B,et al.ROCK inhibitor Y-27632 suppresses dissociation-induced apoptosis of murine prostate stem/progenitor cells and increases their cloning efficiency[J].PLoS One,2011,6(3):e18271.
[22] Koch JC,Tönges L,Barski E,et al.ROCK2 is a major regulator of axonal degeneration,neuronal death and axonal regeneration in the CNS[J].Cell Death Dis,2014,5:e1225.
[23] Stankiewicz TR,Loucks FA,Schroeder EK,et al.Signal transducer and activator of transcription-5 mediates neuronal apoptosis induced by inhibition of Rac GTPase activity[J].J Biol Chem,2012,287(20):16835-16848.

基金

国家自然科学基金资助项目(81471087)。


PDF(2121 KB)
HTML

Accesses

Citation

Detail

段落导航
相关文章

/