环王巴明抑制sonic hedgehog信号通路对氧糖剥夺/再复氧损伤大鼠皮质神经干细胞增殖的影响

成薇, 王莉, 余萍萍, 沈长波, 杨琴

中国药学杂志 ›› 2016, Vol. 51 ›› Issue (8) : 625-629.

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中国药学杂志
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中国药学杂志 ›› 2016, Vol. 51 ›› Issue (8) : 625-629. DOI: 10.11669/cpj.2016.08.007
论 著

环王巴明抑制sonic hedgehog信号通路对氧糖剥夺/再复氧损伤大鼠皮质神经干细胞增殖的影响

  • 成薇, 王莉, 余萍萍, 沈长波, 杨琴*
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Effects of Inhibiting Shh Signaling Pathway with Cyclopamine Pretreatment on Proliferation of Cortical Neural Stem Cells after Oxygen-Glucose Deprivation/Reoxygenation Injury in Rats

  • CHENG Wei, WANG Li, YU Ping-ping, SHEN Chang-bo, YANG Qin*
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摘要

目的 探讨环王巴明预处理抑制sonic hedgehog信号通路对体外氧糖剥夺/再复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤大鼠大脑皮质神经干细胞(neural stem cells,NSCs)增殖的影响。 方法 新生SD大鼠大脑皮质神经干细胞采用悬浮培养法分离纯化。 第3代神经干细胞贴壁培养后氧糖剥夺150 min,复氧培养24 h。实验分为正常组、模型组及环王巴明预处理组。细胞鉴定采用免疫荧光法,细胞活力采用CCK-8法检测,细胞增殖采用BrdU法及流式细胞周期检测,Ptc-1、Smo、Gli-1蛋白表达采用Western blot检测。 结果 悬浮及贴壁培养细胞均高表达巢蛋白。模型组及环王巴明组细胞活力较正常组显著降低。其中,环王巴明组降低更明显(P<0.05)。模型组细胞增殖明显,Ptc-1、Smo、Gli-1蛋白表达上调,而环王巴明组细胞增殖较模型组低,Ptc-1、Smo、Gli-1蛋白表达下调(P<0.05)。 结论 环王巴明预处理能抑制体外氧糖剥夺/再复氧损伤后神经干细胞的增殖,提示Shh信号可能参与损伤后神经干细胞增殖的调控。

Abstract

OBJECTIVE To investigate the effect of inhibitting sonic hedgehog(Shh) signaling pathway with cyclopamine pretreatment on proliferation of rat cortical neural stem cells (NSCs) after oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro. METHODS The suspended culture was used for the isolation and purification of NSCs in neonatal Sprague-Dawley (SD) rats. The third passage NSCs for adherent culture were deprived oxygen and glucose for 150 min and recovered oxygen and glucose for 24 h. There were three groups, including normal, model and cyclopamine pretreatment groups. NSCs were identified with immunofluorescence. CCK-8 assay was used to examine cell viability. The proliferation of NSCs was measured with BrdU assay and flow cytometry cell cycle. Western blot was used to detect the protein expressions of Ptc-1,Smo and Gli-1. RESULTS There were high expression of nestin protein in suspended and adherent cultured cells. The cell vitalities in model and cyclopamine groups were decreased significantly compared with the normal group. Especially, there was less cell vitality in cyclopamine group(P<0.05). There was significantly increased for NSCs proliferation and upregulated for Ptc-1,Smo and Gli-1 proteins in the model group. On the contrary, compared with the model group, NSCs proliferation and the expressions of Ptc-1,Smo and Gli-1 proteins in cyclopamine group were significantly decreased(P<0.05). CONCLUSION Cyclopamine pretreatment can inhibit NSCs proliferation after OGD/R injury. The result suggests that Shh signaling may participate in the regulation of NSCs proliferation after injury.

关键词

环王巴明 / 预处理 / 神经干细胞 / 氧糖剥夺/再复氧 / 增殖 / sonic hedgehog

Key words

cyclopamine / pretreatment / neural stem cells / oxygen-glucose deprivation/reoxygenation / proliferation / sonic hedgehog

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成薇, 王莉, 余萍萍, 沈长波, 杨琴. 环王巴明抑制sonic hedgehog信号通路对氧糖剥夺/再复氧损伤大鼠皮质神经干细胞增殖的影响[J]. 中国药学杂志, 2016, 51(8): 625-629 https://doi.org/10.11669/cpj.2016.08.007
CHENG Wei, WANG Li, YU Ping-ping, SHEN Chang-bo, YANG Qin. Effects of Inhibiting Shh Signaling Pathway with Cyclopamine Pretreatment on Proliferation of Cortical Neural Stem Cells after Oxygen-Glucose Deprivation/Reoxygenation Injury in Rats[J]. Chinese Pharmaceutical Journal, 2016, 51(8): 625-629 https://doi.org/10.11669/cpj.2016.08.007
中图分类号： R965   

参考文献

[1] MATISE M P, WANG H. Sonic hedgehog signaling in the developing CNS:Where it has been and where it is going[J]. Curr Top Dev Biol, 2011, 97:75-117.
[2] FAN C C, YANG Q, CHEN N, et al. Dynamic changes of the expression of sonic hedgehog signaling pathway in the subventricular zone after focal cerebral ischemia in rat [J]. Life Sci Res(生命科学研究),2011,15(6): 524-529.
[3] SIMS J R, LEE S W, TOPALKARA K, et al. Sonic hedgehog regulates ischemia/ hypoxia-induced neural progenitor proliferation[J]. Stroke, 2009, 40(11): 3618-3626.
[4] HERETSCH P, TZAGKAROULAKI L, GIANNIS A. Cyclopamine and hedgehog signaling:Chemistry, biology, medical perspectives [J]. Angew Chem Int Ed Engl, 2010, 49(20): 3418-3427.
[5] DRENKHAHN S K, JACKSON G A, SLUSARZ A, et al. Inhibition of hedgehog/Gli signaling by botanicals: A review of compounds with potential hedgehog pathway inhibitory activities[J]. Curr Cancer Drug Targets, 2013, 13(5): 580-595.
[6] YANG Q,ZENG Z L,XIE P,et al. Identification and cultivation of neural stem cells in neonatal rats in vitro[J]. J Chongqing Med Univ(重庆医科大学学报),2007,32(11): 1134-1137.
[7] BABU H, CLAASEN J H, KANNAN S, et al. A protocol for isolation and enriched monolayer cultivation of neural precursor cells from mouse dentate gyrus[J]. Front Neurosci, 2011, 5:89.
[8] CHEN T, LIU W, CHAO X, QU Y, et al. Neuroprotective effect of osthole against oxygen and glucose deprivation in rat cortical neurons:Involvement of mitogen-activated protein kinase pathway[J]. Neurosci, 2011, 183(2):203-211.
[9] SHEN C B, HUANG J G, ZHANG L L, et al. Effect of resveratrol on oxidative stress of rat primary cortical neurons during different time windows of oxygen-glucose deprivation/reperfusion injury[J]. Acad J Sec Mil Med Univ(第二军医大学学报),2013,34(7): 708-713.
[10] REN J W, FAN C C, CHEN N, et al. Resveratrol pretreatment attenuates cerebral ischemic injury by upregulating expression of transcription factor Nrf2 and HO-1 in rats[J]. Neurochem Res, 2011, 36: 2352-2362.
[11] ZHANG L, CHOPP M, MEIER D H, et al. Sonic hedgehog signaling pathway mediates cerebrolysin-improved neurological function after stroke[J]. Stroke, 2013, 44(7):1965-1972.
[12] ZHANG L L,HUANG J G,SHEN C B, et al. Effect of resveratrol on rat primary cortical neurons during oxygen and glucose deprivation/reperfusion injury[J]. Chin Tradit Pat Med(中成药),2014,36(5): 897-903.
[13] CHOY S W, CHENG S H. Hedgehog signaling[J]. Vitam Horm, 2012, 88:1-23.
[14] LAI K, KASPAR B K, GAGE F H, et al. Sonic hedgehog regulates adult neural progenitor proliferation in vitro and in vivo[J]. Nat Neurosc, 2003, 6(1):21-27.
[15] JI H, MIAO J, ZHANG X, et al. Inhibition of sonic hedgehog signaling aggravates brain damage associated with the down-regulation of Gli1, Ptch1 and SOD1 expression in acute ischemic stroke[J]. Neurosci Lett, 2012,506(1):1-6.
[16] DING X, LI Y, LIU Z, et al. The sonic hedgehog pathway mediates brain plasticity and subsequent functional recovery after bone marrow stromal cell treatment of stroke in mice[J]. J Cereb Blood Flow Metab, 2013,33(7):1015-1024.
[17] JIN Y, RAVIV N, BARNETT A, et al. The shh signaling pathway is upregulated in multiple cell types in cortical ischemia and influences the outcome of stroke in an animal model[J]. PLoS One, 2015,10(4):e0124657.
[18] CHECHNEVA O V, MAYRHOFER F, DAUGHERTY D J, et al. A smoothened receptor agonist is neuroprotective and promotes regeneration after ischemic brain injury[J]. Cell Death Dis,2014,23,5:e1481.
[19] HUANG S S, CHENG H, TANG C M, et al. Anti-oxidative, anti-apoptotic, and pro-angiogenic effects mediate functional improvement by sonic hedgehog against focal cerebral ischemia in rats[J]. Exp Neurol Sep,2013,247: 680-688.

基金

国家自然科学基金面上项目(81071119);国家神经病学临床重点专科建设资助项目(卫办医政函【2012】649号)

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