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  

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

作　　者：阿不拉江·托合提 程秀琴[1] 古丽波斯坦·买买提艾力[1] Abulajiang Tuoheti;CHENG Xiu-qin;Gulibositan Maimaitiaili(Department of Otorhinolaryngology,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xinjiang 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院耳鼻咽喉科,新疆乌鲁木齐830001

出　　处：《局解手术学杂志》2023年第7期567-572,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：新疆维吾尔自治区自然科学基金(2022D01C135);新疆维吾尔自治区人民医院内项目(20200116)。

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(P<0.05),血清中PAD4、CitH3含量升高(P<0.05),且两者呈正相关(R=0.3636)。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(P<0.05),PAD4蛋白、CitH3蛋白、p-P65/P65及IL-6、TNF-αmRNA表达水平升高(P<0.05),血清中PAD4、CitH3、IL-6、TNF-α含量升高(P<0.05);与模型组比较,给药组听性脑干反应阈值降低(P<0.05),PAD4蛋白、CitH3蛋白、p-P65/P65及IL-6、TNF-αmRNA表达水平降低(P<0.05),血清中PAD4、CitH3、IL-6、TNF-α降低(P<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。Objective To investigate the mechanism of action of citrullinated histone H3(CitH3)mediated by arginine deimi-mase 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-αin 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(P<0.05),and the contents of PAD4 and CitH3 in serum increased(P<0.05),and they were positively correlated(R=0.3636).Animal experiments showed that compared with the control group,the threshold of auditory brainstem response in the model group was increased(P<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-αwere increased(P<0.05),the contents of PAD4,CitH3,IL-6 and TNF-αin serum were increased(P<0.05).Compared with the model group,the threshold of auditory brainstem response in drug administration group was decreased(P<0.05),the protien expression levels of PAD4 and CitH3,the level of p-P65/P65 and the

关 键 词：中耳炎 精氨酸脱亚胺酶4 瓜氨酸化组蛋白 脂多糖 

分 类 号：R764.21[医药卫生—耳鼻咽喉科]