Mechanism of PAD4-mediated citrullinated histone H3 in the pathogenesis of otitis media

Abulajiang Tuoheti; CHENG Xiu-qin; Gulibositan Maimaitiaili

doi:10.11659/jjssx.07E022105

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Mechanism of PAD4-mediated citrullinated histone H3 in the pathogenesis of otitis media

Basic Research | 更新时间：2023-07-17

- Mechanism of PAD4-mediated citrullinated histone H3 in the pathogenesis of otitis media
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 32, Issue 7, Pages: 567-572(2023)
- 作者机构：
  
  新疆维吾尔自治区人民医院耳鼻咽喉科，新疆乌鲁木齐 830001
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.07E022105
  CLC： R764.21
- Published：25 July 2023，
  
  Received：27 July 2022，
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阿不拉江·托合提 ,程秀琴,古丽波斯坦·买买提艾力 .PAD4介导的瓜氨酸化组蛋白在中耳炎发病进程中的机制研究[J].局解手术学杂志,2023,32(07):567-572.

Tuoheti Abulajiang,CHENG Xiu-qin,Maimaitiaili Gulibositan.Mechanism of PAD4-mediated citrullinated histone H3 in the pathogenesis of otitis media[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(07):567-572.
阿不拉江·托合提 ,程秀琴,古丽波斯坦·买买提艾力 .PAD4介导的瓜氨酸化组蛋白在中耳炎发病进程中的机制研究[J].局解手术学杂志,2023,32(07):567-572. DOI： 10.11659/jjssx.07E022105.

Tuoheti Abulajiang,CHENG Xiu-qin,Maimaitiaili Gulibositan.Mechanism of PAD4-mediated citrullinated histone H3 in the pathogenesis of otitis media[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(07):567-572. DOI： 10.11659/jjssx.07E022105.

摘要

目的

探究精氨酸脱亚胺酶4（PAD4）介导的瓜氨酸化组蛋白3（CitH3）在中耳炎发病进程中的作用机制。

方法

收集于我院接受手术治疗的慢性中耳炎患者和非中耳炎患者各30例。采用Western blot检测患者耳组织中PAD4、CitH3蛋白表达水平；ELISA检测患者血清中PAD4、CitH3含量；并分析PAD4与CitH3的相关性。将30只BALB/c小鼠分为对照组、模型组和给药组，每组10只。模型组和给药组中耳注射脂多糖（LPS）建模后，给药组每日中耳注射5 mg/kg cl-amidine，模型组和对照组注射等量生理盐水；实验结束后进行听性脑干反应测试；Western blot检测PAD4、CitH3、p-P65、P65蛋白表达水平；qRT-PCR检测IL-6、TNF-α表达水平；ELISA检测血清中PAD4、CitH3、IL-6、TNF-α含量。

结果

与非中耳炎患者比较，慢性中耳炎患者耳组织中PAD4、CitH3蛋白表达水平升高（

0.05），血清中PAD4、CitH3含量升高（

0.05），且两者呈正相关（

0.363 6）。动物实验结果显示，与对照组比较，模型组听性脑干反应阈值升高（

0.05），PAD4蛋白、CitH3蛋白、p-P65/P65及IL-6、TNF-α mRNA表达水平升高（

0.05），血清中PAD4、CitH3、IL-6、TNF-α含量升高（

0.05）；与模型组比较，给药组听性脑干反应阈值降低（

0.05），PAD4蛋白、CitH3蛋白、p-P65/P65及IL-6、TNF-α mRNA表达水平降低（

0.05），血清中PAD4、CitH3、IL-6、TNF-α降低（

0.05）。

结论

PAD4通过介导CitH3激活NF-κB信号通路，促进耳组织炎症反应；抑制PAD4活性可缓解中耳炎症状。

Abstract

Objective

To investigate the mechanism of action of citrullinated histone H3 （CitH3） mediated by arginine deimimase 4 （PAD4） in the pathogenesis of otitis media.

Methods

Thirty patients with chronic otitis media and 30 patients with non-otitis media who received surgical treatment in our hospital were collected.The expression levels of PAD4 and CitH3 in ear tissues of patients were detected by Western blot.The contents of PAD4 and CitH3 in serum were detected by ELISA，and the correlation between PAD4 and CitH3 was analyzed.Thirty BALB/c mice were divided into the control group，model group and drug administration group，with 10 mice in each group. The model group and drug administration group were injected with lipopolysaccharide （LPS） to establish model，then the drug administration group was injected with 5 mg/kg cl-amidine，and the model group and control group were injected with the same dose of normal saline.After the experiment，auditory brainstem response test was performed.Western blot was used to detect the expression levels of PAD4，CitH3，p-P65 and P65；qRT-PCR was used to detect the expression levels of IL-6 and TNF-α. ELISA was used to detect the contents of PAD4，CitH3，IL-6 and TNF-α in serum.

Results

Compared with non-otitis media patients，the protein expression levels of PAD4 and CitH3 in tissues of patients with chronic otitis media increased （

0.05），and the contents of PAD4 and CitH3 in serum increased （

0.05），and they were positively correlated （

=0.363 6）.Animal experiments showed that compared with the control group，the threshold of auditory brainstem response in the model group was increased （

0.05），the protein expression levels of PAD4 and CitH3，the level of p-P65/P65 and the mRNA expression levels of IL-6，TNF-α were increased （

0.05），the contents of PAD4，CitH3，IL-6 and TNF-α in serum were increased （

0.05）.Compared with the model group，the threshold of auditory brainstem response in drug administration group was decreased （

0.05），the protien expression levels of PAD4 and CitH3，the level of p-P65/P65 and the mRNA expression levels of IL-6 and TNF-α were decreased （

0.05），the contents of PAD4， CitH3，IL-6 and TNF-α in serum were decreased （

0.05）.

Conclusion

PAD4 promotes inflammatory responses in ear tissue by mediating CitH3 and activating the NF-κB signaling pathway.Inhibition of PAD4 activity may alleviate symptoms of otitis media disease.

关键词

中耳炎精氨酸脱亚胺酶4瓜氨酸化组蛋白脂多糖

Keywords

otitis mediaarginine deiminase 4citrullinated histone H3lipopolysaccharide

references

Morris PS，Leach AJ.Acute and chronic otitis media［J］.Pediatr Clin North Am，2009，56（6）：1383-1399. doi： 10.1016/j.pcl.2009. 09.007http://dx.doi.org/10.1016/j.pcl.2009.09.007.

Bhutta MF，Thornton RB，Kirkham LS，et al.Understanding the aetiology and resolution of chronic otitis media from animal and human studies［J］.Dis Model Mech，2017，10（11）：1289-1300. doi： 10.1242/dmm.029983http://dx.doi.org/10.1242/dmm.029983.

Tsourouktsoglou TD，Warnatsch A，Ioannou M，et al.Histones，DNA，and citrullination promote neutrophil extracellular trap inflammation by regulating the localization and activation of TLR4［J］.Cell Rep，2020，31（5）：107602. doi： 10.1016/j.celrep.2020.107602http://dx.doi.org/10.1016/j.celrep.2020.107602.

Makrygiannakis D，af Klint E，Lundberg IE，et al.Citrullination is an inflammation-dependent process［J］.Ann Rheum Dis，2006，65（9）：1219-1222. doi： 10.1136/ard.2005.049403http://dx.doi.org/10.1136/ard.2005.049403.

Morita S，Nakamaru Y，Fukuda A，et al.The quantification of extracellular trap cell death-derived products as diagnostic biomarkers for otitis media with antineutrophil cytoplasmic antibody-associated vasculitis and eosinophilic otitis media［J］.Otol Neurotol，2022，43（3）：e337-e343. doi： 10.1097/MAO.0000000000003431http://dx.doi.org/10.1097/MAO.0000000000003431.

Wang Y，Li M，Stadler S，et al.Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formation［J］.J Cell Biol，2009，184（2）：205-213. doi： 10.1083/jcb. 200806072http://dx.doi.org/10.1083/jcb.200806072.

Yang C，Dong ZZ，Zhang J，et al.Peptidylarginine deiminases 4 as a promising target in drug discovery［J］.Eur J Med Chem，2021，226：113840. doi： 10.1016/j.ejmech.2021http://dx.doi.org/10.1016/j.ejmech.2021.

童兴科，孙永东，向小红.基于水通道蛋白清窍胶囊（Qingqiao Capsule）治疗分泌性中耳炎小鼠模型机制的研究［J］.世界最新医学信息文摘，2018，18（72）：20-21. doi：10.19613/j.cnki.1671-3141.2018.72.007http://dx.doi.org/10.19613/j.cnki.1671-3141.2018.72.007.

Hang A，Brietzke SE.Otitis media： epidemiology and management［J］.Infect Disord Drug Targets，2012，12（4）：261-266. doi： 10.2174/187152612801319339http://dx.doi.org/10.2174/187152612801319339.

Lasminingrum L，Mahdiani S，Dermawan A，et al.Impact of the COVID-19 pandemic on the volume of chronic suppurative otitis media surgeries： a cohort retrospective study ［J］.Ann Med Surg （Lond），2022，79：103921. doi： 10.1016/j.amsu.2022http://dx.doi.org/10.1016/j.amsu.2022.

Safari D，Wahyono DJ，Tafroji W，et al.Serotype distribution and antimicrobial resistance profile of haemophilus influenzae isolated from school children with acute otitis media［J］.Int J Microbiol，2022，2022：5391291. doi： 10.1155/2022/5391291http://dx.doi.org/10.1155/2022/5391291.

Thålin C，Aguilera K，Hall NW，et al.Quantification of citrullinated histones： development of an improved assay to reliably quantify nucleosomal H3Cit in human plasma［J］.J Thromb Haemost，2020，18（10）：2732-2743. doi： 10.1111/jth.15003http://dx.doi.org/10.1111/jth.15003.

Li M，Lin C，Deng H，et al.A novel peptidylarginine deiminase 4 （PAD4） inhibitor BMS-P5 blocks formation of neutrophil extracellular traps and delays progression of multiple myeloma［J］.Mol Cancer Ther，2020，19（7）：1530-1538. doi： 10.1158/1535-7163http://dx.doi.org/10.1158/1535-7163.

Matar GM，Sidani N，Fayad M，et al.Two-step PCR-based assay for identification of bacterial etiology of otitis media with effusion in infected Lebanese children［J］.J Clin Microbiol，1998，36（5）：1185-1188. doi： 10.1128/JCM.36.5.1185-1188http://dx.doi.org/10.1128/JCM.36.5.1185-1188.

Schousboe LP，Ovesen T，Eckhardt L，et al.How does endotoxin trigger inflammation in otitis media with effusion？［J］.Laryngoscope，2001，111（2）：297-300. doi： 10.1097/00005537-200102000-00020http://dx.doi.org/10.1097/00005537-200102000-00020.

Subramanian V，Knight JS，Parelkar S，et al.Design，synthesis，and biological evaluation of tetrazole analogs of Cl-amidine as protein arginine deiminase inhibitors［J］.J Med Chem，2015，58（3）：1337-1344. doi： 10.1021/jm501636xhttp://dx.doi.org/10.1021/jm501636x.

Kim BG，Choi DY，Kim MG，et al.Effect of angiogenesis and lymphangiogenesis in diesel exhaust particles inhalation in mouse model of LPS induced acute otitis media［J］.Front Cell Infect Microbiol，2022，12：824575. doi： 10.3389/fcimb.2022.824575http://dx.doi.org/10.3389/fcimb.2022.824575.

Serban R，Filip C，Radulescu LM，et al.IL-1α，IL-6 and IL-8 serum values in patients with chronic suppurative otitis media［J］.Exp Ther Med，2021，22（5）：1226. doi： 10.3892/etmhttp://dx.doi.org/10.3892/etm.

Fujita K， Tokuda H， Yamamoto N， et al. Incretins amplify TNF-α- stimulated IL-6 synthesis in osteoblasts： Suppression of the IκB/NF-κB pathway［J］. Int J Mol Med， 2017， 39（4）：1053-1060. doi： 10.3892/ijmm.2017.2892http://dx.doi.org/10.3892/ijmm.2017.2892.

Barzilaia A，Dekel B，Dagan R，et al.Middle ear effusion IL-6 concentration in bacterial and non-bacterial acute otitis media［J］.Acta Paediatr，2000，89（9）：1068-1071. doi： 10.1080/713794564http://dx.doi.org/10.1080/713794564.

Zhao X，Gu C，Wang Y.PAD4 selective inhibitor TDFA protects lipopolysaccharide-induced acute lung injury by modulating nuclear p65 localization in epithelial cells［J］.Int Immunopharmacol，2020，88：106923. doi： 10.1016/j.intimp.2020.106923http://dx.doi.org/10.1016/j.intimp.2020.106923.

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