白藜芦醇对NIH3T3细胞Shh信号通路的影响

郭霜 廖鸿雁 刘杰 唐凡人 杨琴*

doi:10.16098/j.issn.0529-1356.2018.02.007

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解剖学报 ›› 2018, Vol. 49 ›› Issue (2) : 179-184. DOI: 10.16098/j.issn.0529-1356.2018.02.007

细胞和分子生物学

白藜芦醇对NIH3T3细胞Shh信号通路的影响

郭霜廖鸿雁刘杰唐凡人杨琴*

作者信息 +

Effects of resveratrol on Shh signaling pathway of NIH3T3 cells

GUO Shuang LIAO Hong-yan LIU Jie TANG Fan-ren YANG Qin*

Author information +

文章历史 +

摘要

目的探讨白藜芦醇对NIH3T3细胞Shh信号通路的影响。方法实验分为对照组和白藜芦醇组。白藜芦醇处理NIH3T3细胞24 h。CCK-8法测定细胞活力，免疫荧光法检测初级纤毛蛋白（Ac-tu）、Smo和Gli-1的表达，Western blotting法检测Shh、Ptc、Smo、Gli-1蛋白的表达。结果 1. 0.5和1 μmol/L白藜芦醇组（0.679±0.047, 0.774±0.054）细胞活力分别较对照组（0.585±0.039）明显增强（P＜0.05），其中以1 μmol/L白藜芦醇组最强。之后随白藜芦醇浓度（10、20、40、80 μmol/L）的增高，细胞活力逐渐降低（0.428±0.043, 0.395±0.031, 0.373±0.017, 0.361±0.016），且均较对照组明显减弱（P＜0.05），而0.1 μmol/L（0.602±0.065）和5 μmol/L组（0.556±0.041）细胞活力与对照组比较差异无显著性（P>0.05）。2. NIH3T3细胞表达Ac-tu、Shh、Ptc-1、Smo、Gli-1从蛋白，有1根初级纤毛，Smo和Gli-1蛋白位于细胞质。白藜芦醇处理24h时，Gli-1从细胞质进入细胞核，Smo从细胞质进入初级纤毛，且Shh（0.756±0.659比0.441±0.769，P＜0.05）、Ptc-1（0.655±0.347比0.351±0.026，P＜0.05）、Smo（0.779±0.064比0.451±0.035，P＜0.05）和Gli-1（0.856±0.044比0.560±0.058，P＜0.05）蛋白表达较对照组高。结论白藜芦醇可能通过激活Shh信号通路增强NIH3T3细胞的活力。

Abstract

Objective To investigate the effect of resveratrol on Shh signaling pathway of NIH3T3 cells in vitro. Methods NIH3T3 cells were divided into the control and resveratrol groups. The cells were cultured with resveratrol for 24 hours. Cell viability was detected with CCK-8 assay．Immunofluorescence measured the expressions of Ac-tu, Smo, and Gli-1. Western blotting assayed the expressions of Shh, Ptc-1, Smo and Gli-1 proteins. Results Compared with the control group（0.585±0.039）, 0.5（0.679±0.047, P＜0.05 and 1.0 μmol/L（0.774±0.054, P＜0.05 resveratrol significantly enhanced the viability of NIH3T3 cells, and the peak was 1.0 μmol/L. On the contrary, 10, 20, 40, 80 μmol/L resveratrol significantly decreased the viability of NIH3T3 cells（0.428±0.043, 0.395±0.031, 0.373±0.017, 0.361±0.016, respectively). However, 0.1（0.602±0.065）and 5 μmol/L（0.556±0.041）resveratrol did not affect the viability（P>0.05）. NIH3T3 cells had one primary cilia and expressed the proteins of Ac-Tu, Shh, Ptc-1, Smo and Gli-1.Smo and Gli-1 proteins were located in the cytoplasm. At 24 hours for resveratrol treat, Gli-1 protein translocated into the nucleus from cytoplasm and Smo protein entered the primary cilia from the cytoplasm. Expressions of Shh（0.756±0.659 vs 0.441±0.769，P＜0.05,Ptc-1（0.655±0.347 vs 0.351±0.026，P＜0.05,Smo（0.779±0.064 vs 0.451±0.035，P＜0.05 and Gli-1（0.856±0.044 vs 0.560±0.058，P＜0.05 proteins significantly compared with the control group. Conclusion Resveratrol can enhance viability of NIH3T3 cells via activating Shh signaling pathway.

导出引用

郭霜廖鸿雁刘杰唐凡人杨琴. 白藜芦醇对NIH3T3细胞Shh信号通路的影响[J]. 解剖学报. 2018, 49(2): 179-184 https://doi.org/10.16098/j.issn.0529-1356.2018.02.007

GUO Shuang LIAO Hong-yan LIU Jie TANG Fan-ren YANG Qin. Effects of resveratrol on Shh signaling pathway of NIH3T3 cells[J]. Acta Anatomica Sinica. 2018, 49(2): 179-184 https://doi.org/10.16098/j.issn.0529-1356.2018.02.007

参考文献

［1］ Baur JA, Sinclair DA. Therapeutic potential of resveratrol: the in vivo evidence［J］. Nat Rev Drug Discov, 2006, 5(6):493-506.

［2］Diaz-Gerevini GT, Repossi G, Dain A, et al. Beneficial action of resveratrol: How and why［J］? Nutrition 2016, 32(2):174-178.

［3］Bastianetto S, Ménard C, Quirion R. Neuroprotective action of resveratrol［J］. Biochim Biophys Acta, 2015, 1852(6):1195-1201.

［4］Ren JW, Yang Q. Protective mechanism of resveratrol on cerebral ischemia / reperfusion injury via ameliorating oxidative stress ［J］. Chinese Pharmacological Bulletin,2011，27(4) (12):448-451. (in Chinese)

任俊伟, 杨琴. 白藜芦醇对脑缺血/再灌注氧化应激损伤保护机制的研究进展［J］. 中国药理学通报, 2011，27(4): 448-451.

［5］Ren J, Fan 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(12): 2352-2362.

［6］Yu P, Wang L, Tang F, et al. Resveratrol pretreatment decreases ischemic injury and improves neurological function via sonic hedgehog signaling after stroke in rats［J］. Mol Neurobiol, 2017，54(1): 212-226.

［7］Cheng W, Yu P, Wang L, et al. Sonic hedgehog signaling mediates resveratrol to increase proliferation of neural stem cells after oxygen-glucose deprivation/re oxygenation injury in vitro［J］. Cell Physiol Biochem, 2015，35(5): 2019-2032.

［8］Shen C, Cheng W, Yu P, et al. Resveratrol pretreatment attenuates injury and promotes proliferation of neural stem cells following oxygen-glucose deprivation/reoxygenation by upregulating the expressions of Nrf2, HO-1 and NQO1 in vitro［J］. Mol Med Rep, 2016, 14(4):3646-3654.

［9］Yu PP，Wang L，Tang FR，et al． Effect of resveratrol on activation of astrocyte after cerebral ischemia /reperfusion injury in rats［J］．Chinese Pharmacological Bulletin, 2015，31 (9): 1228-1233．(in Chinese)

余萍萍, 王莉, 唐凡人, 等. 白藜芦醇对大鼠脑缺血/再灌注损伤后星形胶质细胞活化的影响［J］. 中国药理学通报, 2015，31(9): 1228-1233.

［10］Yu PP，Wang L，Tang FR，et al．Effect of resveratrol on expression of synaptophysin after cerebral ischemia /reperfusion injury in rats［J］．Acta Anatomica Sinica, 2016，47(1): 12-17. (in Chinese)

余萍萍, 王莉, 唐凡人, 等. 白藜芦醇对大鼠脑缺血再灌注损伤后突触素表达的影响［J］. 解剖学报, 2016，47(1): 12-17.

［11］Zhang LL， Huang JG，Shen ChB，et al． Effect of resveratrol on rat primary cortical neurons during oxygen and glucose deprivation /reperfusion injury［ J］．Chinese Traditional Patent Medicine, 2014，36(5): 897-903．(in Chinese)

张黎黎, 黄家贵, 沈长波, 等. 白藜芦醇预处理对缺糖缺氧再灌注大鼠原代皮质神经元的保护作用［J］. 中成药, 2014，36(5): 897-903.

［12］Tang FR，Yu PP，Wang L，et al．Effects of resveratrol pretreatment on neurite growth of primary cortical neurons after oxygen-glucose deprivation /reperfusion injury in rats［J］．Acta Anatomica Sinica，2017，48(1): 1-6. (in Chinese)

唐凡人, 余萍萍, 王莉, 等. 白藜芦醇预处理对大鼠皮质神经元氧糖剥夺/再复氧损伤后神经元突起生长的影响［J］. 解剖学报, 2017，48(1): 1-6.

［13］Shen ChB， Huang JG， Zhang LL， et al． Effect of resveratrol on oxidative stress of rat primary cortical neurons during different time windows of oxygen-glucose deprivation /reperfusion injury［J］．Academic Journal of Second Military Medical University，2013， 34(7): 708-713． (in Chinese)

沈长波，黄家贵，张黎黎，等. 白藜芦醇在缺血缺氧再灌注损伤不同时间窗皮质神经元氧化应激的影响［J］. 第二军医大学学报，2013, 34（7）：708-713.

［14］Patel SS, Tomar S, Sharma D, et al. Targeting sonic hedgehog signaling in neurological disorders［J］. Neurosci Biobehav Rev, 2017，74(Pt A): 76-97.

［15］Zhang L, Chopp M, Meier DH, et al. Sonic hedgehog signaling pathway mediates cerebrolysin improved neurological function after stroke［J］. Stroke, 2013, 44(7):1965-1972.

［16］Huang SS, Cheng H, Tang CM, 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.

［17］Chechneva OV, Mayrhofer F, Daugherty DJ, et al. A Smoothened receptor agonist is neuroprotective and promotes regeneration after ischemic brain injury［J］. Cell Death Dis, 2014, 5(10):e1481.

［18］Huang JG, Shen CB, Wu WB, et al. Primary cilia mediate sonic hedgehog signaling to regulate neuronal-like differentiation of bone mesenchymal stem cells for resveratrol induction in vitro［J］. J Neurosci Res, 2014，92(5): 587-596.

［19］Park HR, Kong KH, Yu BP, et al. Resveratrol inhibits the proliferation of neural progenitor cells and hippocampal neurogenesis［J］. J Biol Chem, 2012, 287(51):42588-42600.

［20］Leong CW, Wong CH, Lao SC, et al. Effect of resveratrol on proliferation and differentiation of embryonic cardiomyoblasts［J］. Biochem Biophys Res Commun, 2007,360(1):173-180.

［21］Huang JG, Ren JW, Yang Q. Study on the correlation between primary cilia and Hedgehog signaling pathway［J］．Chinese Journal of Anatomy, 2011，34(4): 552-555. (in Chinese)

黄家贵，任俊伟，杨琴. 初级纤毛与Hedgehog信号通路相关性研究［J］.解剖学杂志，2011，34（4）：552-555.

［22］Walz G. Role of primary cilia in non-dividing and post-mitotic cells［J］.Cell Tissue Res, 2017,369(1):11-25.