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中华口腔医学研究杂志(电子版) ›› 2016, Vol. 10 ›› Issue (02) : 112 -119. doi: 10.3877/cma.j.issn.1674-1366.2016.02.006

所属专题: 文献

基础研究

前扣带回皮质层中牙移动疼痛相关蛋白质组学研究
赵转浓1, 刘楚峰2, 孙晓卫1, 张灵超2, 曹阳1,()   
  1. 1. 510055 广州,中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室
    2. 510280 广州,广东省口腔医院·南方医科大学附属口腔医院正畸科
  • 收稿日期:2015-11-12 出版日期:2016-04-01
  • 通信作者: 曹阳
  • 基金资助:
    国家自然科学基金(81170990、81200813)

Proteomic analysis related with pain of tooth movement in the anterior cingulate cortex

Zhuannong Zhao1, Chufeng Liu2, Xiaowei Sun1, Lingchao Zhang2, Yang Cao1,()   

  1. 1. Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
    2. Department of Orthodontics, Guangdong Provincial Stomatological Hospital, Affiliated Stomatological Hospital of Southern Medical University, Guangzhou 510280, China
  • Received:2015-11-12 Published:2016-04-01
  • Corresponding author: Yang Cao
  • About author:
    Corresponding author: Cao Yang, Email:
引用本文:

赵转浓, 刘楚峰, 孙晓卫, 张灵超, 曹阳. 前扣带回皮质层中牙移动疼痛相关蛋白质组学研究[J]. 中华口腔医学研究杂志(电子版), 2016, 10(02): 112-119.

Zhuannong Zhao, Chufeng Liu, Xiaowei Sun, Lingchao Zhang, Yang Cao. Proteomic analysis related with pain of tooth movement in the anterior cingulate cortex[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2016, 10(02): 112-119.

目的

研究正畸牙移动中前扣带回皮质层(ACC)的蛋白质组学表达,筛选牙移动疼痛相关的目标蛋白,探讨ACC在疼痛传导和形成中的作用。

方法

30只雄性SD大鼠建立实验性牙移动模型后,根据行为学观测结果分组。实验组进行加力,对照组仅进行乙醚麻醉,24 h后进行行为学观测并取ACC组织,根据行为学观测结果选取4标样本进行蛋白质组学研究。ACC组织在提取蛋白并胰酶消化后,经串联质谱标记(TMT)试剂标记,进行液相色谱-串联质谱(LC-MS/MS)检测,并对差异表达蛋白进行生物信息学分析,进行基因本体(GO)注释,GO功能聚类分析,基因和基因组学京都百科全书(KEGG)代谢通路注释及功能聚类分析。

结果

本实验共鉴定并定量3374种蛋白,其中表达上调蛋白12种、表达下调蛋白109种。GO注释和功能聚类分析中从生物过程、细胞组成、分子功能三个方面阐述了大鼠在实验性牙移动应力下发生的一系列应激反应及代谢调节。KEGG注释及功能聚类分析表明与炎症相关通路花生四烯酸通路及JAK-STAT信号通路表达下调,且差异具有统计学意义。花生四烯酸通路的前列腺素E合成酶(PGES)、谷胱甘肽过氧化物酶1(GPX1)、环氧化物水解酶2(EPHX2),JAK-STAT信号通路的信号转导和转录因子1(STAT1)、STAT2显著下调,可能与炎症性疼痛的发生、发展密切相关。

结论

TMT技术是组织蛋白质组学研究的有效方法。ACC在牙移动疼痛形成和调节中发挥重要作用,其中花生四烯酸通路、JAK-STAT信号通路可能是牙移动疼痛的关键信号通路。本实验为进一步探讨牙移动疼痛关键蛋白提供了重要的实验依据。

Objective

To illucidate the proteomic expression in the anterior cingulate cortex (ACC) after experimental tooth movement and screening the significant proteins related with orthodontic pain, further investigate the role ACC plays in orthodontic pain.

Methods

In this research, 30 male Sprague-Dawley rats (250~ 300 g) were applied with orthodontic appliance. They were divided into two groups and selected for further experiment according to behavior assessment. After tandem mass tag (TMT) labeling, the protein extracted from ACC was detected with liquid chromatography-tandem mass spectrometry (LC-MS/MS) . Then bioinformatic analysis of the differentially expressed proteins was carried out, including Gene Ontology (GO) annotation, GO functional enrichment clustering, and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation and functional enrichment clustering.

Results

In total, 3374 proteins were identified and quantified, among which 109 proteins were significantly down-regulated, while 12 proteins were up-regulated. GO annotation and functional enrichment clustering showed the posttraumatic stress response and biochemical metabolism regulation happened in ACC following orthodontic tooth movement stress from three categories: biological process, cellular component and molecular function. KEGG annotation and functional enrichment clustering results showed that Arachidonic acid metabolism and JAK-STAT signaling pathway which were relevant to inflammatory pain were significantly down-regulated. 5 proteins were found closely related with the inflammary orthodontic pain, including prostaglandin E synthase (PGES) , glutathione peroxidase 1 (GPX1) , bifunctional epoxide hydrolase 2 (EPHX2) in the Arachidonic acid metabolism, as well as signal transducer and activator of transcription 1 (STAT1) , signal transducer and activator of transcription 2 (STAT2) in the JAK-STAT signaling pathway.

Conclusions

TMT technology was proved an effective method evaluating proteomic expression. ACC plays a significant role in the orthodontic pain. Arachidonic acid metabolism and JAK-STAT signaling pathway are possible critical signaling pathways involved in tooth movement.

图1 SD大鼠实验性牙移动模型
表1 与对照组A相比差异表达蛋白数目(种)
图2 基因本体注释结果
图3 基因本体聚类分析
图4 KEGG富集分析
图5 KEGG聚类分析
图6 关键蛋白质的串联质谱图(MS/MS)
[1]
Krishnan V. Orthodontic pain:from causes to management-a review[J]. Eur J Orthod,2007,29(2):170-179.
[2]
Erdinç AM, Dinçer B. Perception of pain during orthodontic treatment with fixed Appliances[J]. Eur J Orthod,2004,26(1):79-85.
[3]
Tecco S, D′attilio M, Tetè S,et al. Prevalence and type of pain during conventional and self-ligating orthodontic treatment[J]. Eur J Orthod,2009,31(4):380-384.
[4]
Scheurer PA, Firestone AR, Bürgin WB. Perception of pain as a result of orthodontic treatment with fixed appliances[J]. Eur J Orthod,1996,18(4):349-357.
[5]
Scott P, Sherriff M, Dibiase AT,et al. Perception of discomfort during initial orthodontic tooth alignment using a self-ligating or conventional bracket system:a randomized clinical trial[J]. Eur J Orthod,2008,30(3):227-232.
[6]
Abdelrahman RSh, Al-nimri KS, Al Maaitah EF. Pain experience during initial alignment with three types of nickel-titanium archwires:a prospective clinical trial[J]. Angle Orthod,2015,85(6):1021-1026.
[7]
Sandhu SS, Sandhu J. A randomized clinical trial investigating pain associated with superelastic nickel-titanium and multistranded stainless steel archwires during the initial leveling and aligning phase of orthodontic treatment[J]. J Orthod,2013,40(4):276-285.
[8]
Jian F, Lai F, Fumess S,et al. Initial arch wires for tooth alignment during orthodontic treatment with fixed appliances[J/OL]. Cochrane Database Syst Rev,2013[2016-03-11]. published online ahead of print April 30,2013].

URL    
[9]
Čelar A, Schedlberger M, Dörfler P,et al. Systematic review on self-ligating vs. conventional brackets:initial pain,number of visits,treatment time[J]. J Orofac Orthop,2013,74(1):40-51.
[10]
Yamashiro T, Satoh K, Nakagawa K,et al. Expression of Fos in the rat forebrain following experimental tooth movement[J]. J Dent Res,1998,77(11):1920-1925.
[11]
Yamashiro T, Nakagawa K, Satoh K,et al. C-fos expression in the trigeminal sensory complex and pontine parabrachial areas following experimental tooth movement[J]. Neuroreport,1997,8(9-10):2351-2353.
[12]
Magdalena CM, Navarro VP, Park DM,et al. C-fos expression in rat brain nuclei following incisor tooth movement[J]. J Dent Res,2004,83(1):50-54.
[13]
Yamashiro T, Kabuto H, Fukunaga T,et al. Medullary monoamine levels during experimental tooth movement[J]. Brain Res,2000,878(1-2):199-203.
[14]
Johansen JP, Fields HL, Manning BH. The affective component of pain in rodents:direct evidence for a contribution of the anterior cingulate cortex[J]. Proc Natl Acad Sci U S A,2001,98(14):8077-8082.
[15]
Lee DE, Kim SJ, Zhuo M. Comparison of behavioral responses to noxious cold and heat in mice[J]. Brain Res,1999,845(1):117-121.
[16]
Hutchison WD, Davis KD, Lozano AM,et al. Pain-related neurons in the human cingulate cortex[J]. Nat Neurosci,1999,2(5):403-405.
[17]
Xu H, Wu LJ, Wang H,et al. Presynaptic and postsynaptic amplifications of neuropathic pain in the anterior cingulate cortex[J]. J Neurosci,2008,28(29):7445-7453.
[18]
Rainville P, Duncan GH, Price DD,et al. Pain affect encoded in human anterior cingulate but not somatosensory cortex[J]. Science,1997,277(5328):968-971.
[19]
Petrovic P, Kalso E, Petersson KM,et al. Placebo and opioid analgesia—imaging a shared neuronal network[J]. Science,2002,295(5560):1737-1740.
[20]
Evans C, Noirel J, Ow SY,et al. An insight into iTRAQ:where do we stand now?[J]. Anal Bioanal Chem,2012,404(4):1011-1027.
[21]
Zieske LR. A perspective on the use of iTRAQTM reagent technology for protein complex and profiling studies[J]. J Exp Bot,2006,57(7):1501-1508.
[22]
Chahrour O, Cobice D, Malone J. Stable isotope labelling methods in mass spectrometry-based quantitative proteomics[J]. J Pharm Biomed Anal,2015(113):2-20
[23]
Yang Z, Wang Y, Luo W,et al. Trigeminal expression of N-methyl-D-aspartate receptor subunit 1 and behavior responses to experimental tooth movement in rats[J]. Angle Orthod,2009,79(5):951-957.
[24]
Yang Z, Cao Y, Wang Y,et al. Behavioural responses and expression of P2X3 receptor in trigeminal ganglion after experimental tooth movement in rats[J]. Arch Oral Biol,2009,54(1):63-70.
[25]
Yang Z, Luo W, Hou JQ,et al. Development of a behavior model of pain induced by experimental tooth movement in rats[J]. Eur J Oral Sci,2009,117(4):380-384.
[26]
Xin YZ, Liu XY, Cao Y. Up-regulation of PKMζ expression in the anterior cingulate cortex following experimental tooth movement in rats[J]. Arch Oral Biol,2014,59(7):749-755.
[27]
陈宇,辛隐子,刘楚峰,等.不同正畸力值作用下大鼠脑前扣带回皮质层蛋白激酶Mζ的表达[J].中华口腔医学杂志,2014,49(12):748-752.
[28]
Imig JD, Hammock BD. Soluble epoxide hydrolase as a therapeutic target for cardiovascular diseases[J]. Nat Rev Drug Discov,2009,8(10):794-805.
[29]
Busch-Dienstfertig M, González-rodriguez S. IL-4,JAK-STAT signaling,and pain[J]. JAKSTAT,2013,2(4):e27638.
[30]
Zhuo M. Molecular mechanisms of pain in the anterior cingulate cortex[J]. J Neurosci Res,2006,84(5):927-933.
[31]
Vogt BA. Pain and emotion interactions in subregions of the cingulate gyrus[J]. Nat Rev Neurosci,2005,6(7):533-544.
[32]
Likhtik E, Pelletier JG, Paz R,et al. Prefrontal control of the amygdala[J]. J Neurosci,2005,25(32):7429-7437.
[33]
Kong J, Tu P, Zyloney C,et al. Intrinsic functional connectivity of the periaqueductal gray,a resting fMRI study[J]. Behav Brain Res,2010,211(2):215-219.
[34]
Qiao H, Gao Y, Zhang C,et al. Increased expression of TRPV1 in the trigeminal ganglion is involved in orofacial pain during experimental tooth movement in rats[J]. Eur J Oral Sci,2015,123(1):17-23.
[35]
Hasegawa M, Kondo M, Suzuki I,et al. ERK is involved in tooth-pressure-induced Fos expression in Vc neurons[J]. J Dent Res,2012,91(12):1141-1146.
[36]
Gao Y, Duan Y. Increased COX2 in the trigeminal nucleus caudalis is involved in orofacial pain induced by experimental tooth movement[J]. Anat Rec(Hoboken),2010,293(3):485-491.
[37]
Fujiyoshi Y, Yamashiro T, Deguchi T,et al. The difference in temporal distribution of c-Fos immunoreactive neurons between the medullary dorsal horn and the trigeminal subnucleus oralis in the rat following experimental tooth movement[J]. Neurosci Lett,2000,283(3):205-208.
[38]
Hiroshima K, Maeda T, Hanada K,et al. Temporal and spatial distribution of Fos protein in the parabrachial nucleus neurons during experimental tooth movement of the rat molar[J]. Brain Res,2001,908(2):161-173.
[39]
Yamashiro T, Fukunaga T, Kabuto H,et al. Activation of the bulbospinal serotonergic system during experimental tooth movement in the rat[J]. J Dent Res,2001,80(9):1854-1857.
[40]
Balam TA, Yamashiro T, Zheng L,et al. Experimental tooth movement upregulates preproenkephalin mRNA in the rat trigeminal nucleus caudalis and oralis[J]. Brain Res,2005,1036(1):196-201.
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