慢性鼻窦炎伴鼻息肉患者血清OPN、BAFF、25-（OH）D3的变化及对息肉组织分型的鉴别价值探讨

摘要 目的：探讨慢性鼻窦炎伴鼻息肉（CRSwNP）患者血清骨桥蛋白（OPN）、B细胞活化因子（BAFF）、25羟基维生素D3[25-（OH）D3]水平及其对息肉组织分型的鉴别价值。方法：选取2019年6月-2021年6月于延安大学附属医院治疗的87例CRSwNP患者为CRSwNP组,根据息肉组织嗜酸粒细胞浸润程度将其分为嗜酸性CRSwNP患者（ECRSwNP组,n=41）和非嗜酸性CRSwNP患者（nECRSwNP组,n=46）。选取同期于延安大学附属医院治疗的慢性鼻窦炎不伴鼻息肉患者（CRSsNP组,n=60）和健康体检者（对照组,n=60）。比较CRSwNP组、CRSsNP组和对照组患者血清OPN、BAFF、25-（OH）D3水平。采用多因素Logistic回归分析ECRSwNP发生的影响因素,采用受试者工作特征（ROC）曲线评估各指标对CRSwNP息肉组织分型的鉴别价值。结果：CRSwNP组、CRSsNP组血清OPN、BAFF水平高于对照组,25-（OH）D3水平低于对照组,且CRSwNP组血清OPN、BAFF水平高于CRSsNP组,25-（OH）D3水平低于CRSsNP组（P<0.05）。ECRSwNP组变应性鼻炎史、哮喘病史患者占比、外周血嗜酸粒细胞数量及比例、血清总IgE浓度、血清OPN及BAFF水平高于nECRSwNP组（P<0.05）,而血清25-（OH）D3水平低于nECRSwNP组（P<0.05）。多因素Logistic回归分析显示：外周血嗜酸粒细胞数量及比例、血清总IgE浓度、血清OPN及BAFF水平升高是ECRSwNP发生的危险因素（P<0.05）,25-（OH）D3水平升高是ECRSwNP的保护因素（P<0.05）。血清OPN、BAFF、25-（OH）D3对CRSwNP息肉组织分型单独检测的ROC曲线下面积（AUC）分别为：0.789、0.800、0.817,联合检测鉴别CRSwNP患者息肉组织分型的AUC为0.900,灵敏度、特异度分别为0.860、0.827,联合检测鉴别价值更高（P＜0.05）。结论：血清OPN、BAFF和25-（OH）D3水平在不同息肉组织分型中具有差异性,CRSwNP患者血清OPN和BAFF水平升高是ECRSwNP发生的危险因素,25-（OH）D3水平升高是保护因素,联合检测辅助临床鉴别CRSwNP息肉组织类型的价值更高。     

Calpeptin通过抑制GATA3减轻变应性鼻炎大鼠炎症

摘要 目的：本研究旨在评估钙蛋白酶抑制剂calpeptin减轻变应性鼻炎大鼠炎症的作用并探讨其机制。方法：将20只雄性SD大鼠采用数字表法随机分为4组：正常组（Normal）、变应性鼻炎组（AR）、地塞米松干预AR组（DXMS+AR）、calpeptin干预AR组（Calpeptin+AR）。造模成功后,对大鼠AR症状进行行为学评分,对大鼠鼻黏膜组织切片以HE和PAS染色法观察鼻黏膜病理改变;对大鼠外周血以ELISA法检测总IgE、IL-4、IL-13水平;对大鼠鼻黏膜组织以免疫蛋白印迹法检测GATA3蛋白表达水平。单因素方差分析进行多组间比较,LSD- t检验进行组间两两比较。结果：与Normal组相比,AR组大鼠的鼻部过敏症状、鼻黏膜嗜酸粒细胞计数及外周血总IgE水平均升高,Calpeptin与地塞米松均能减轻气道炎症,减少嗜酸性粒细胞浸润,降低血清中OVA诱导的IgE的生成。探讨机制发现,酶联免疫吸附试验检测Th2细胞因子,与Normal组比较,AR组血清IL-4、IL-13水平均升高（P＜0.05）,而Calpeptin与地塞米松均能降低血清IL-4、IL-13水平（P＜0.05）。免疫蛋白印迹法检测大鼠鼻黏膜GATA3蛋白表达水平显示,与Normal组比较,AR组鼻黏膜 GATA3表达升高（P＜0.05）,而Calpeptin与地塞米松组鼻黏膜 GATA3表达均下降（P＜0.05）。结论：腹腔注射calpeptin能够减轻变应性鼻炎大鼠局部和全身过敏反应,其机制可能下调GATA3表达,影响Th2细胞的分化及细胞因子的分泌有关。     

稳定期慢性阻塞性肺病患者血嗜酸性粒细胞水平与炎症表型相关性研究

摘要 目的：探究稳定期慢性阻塞性肺病患者血嗜酸性粒细胞水平与炎症表型相关性研究。方法：选择淮安市第一人民医院（我院）2017年5月至2020年3月诊治的稳定期慢性阻塞性肺病患者280例,按照外周血嗜酸性粒细胞计数分为3组,嗜酸性粒细胞＜100为A组86例,嗜酸性粒细胞100~300为B组102例,嗜酸性粒细胞＞300为C组92例。同时纳入同期进行体检的健康者100例为对照组。结果：A组、B组和C组的FeNO显著高于对照组,且C组>B组>A组（P<0.05）;FEV₁占预测值和FEV₁/FVC 显著低于对照组（P<0.05）;A组、B组和C组的FEV₁占预测值和FEV1/FVC对比无差异（P>0.05）。A组、B组和C组的CRP、IL-6、IL-8和TNF-α显著高于对照组（P<0.05）,且C组>B组>A组（P>0.05）。经相关性分析发现,A组和B组患者的嗜酸性粒细胞计数与FeNO、CRP、IL-6、IL-8和TNF-α、FEV₁占预测值和FEV₁/FVC无明显相关性（P>0.05）。C组的嗜酸性粒细胞计数与FeNO、CRP、IL-6、IL-8和TNF-α呈正相关（P<0.05）,与FEV₁/FVC%呈负相关（P<0.05）;与FEV₁占预测值无显著相关性（P>0.05）。结论：稳定期慢性阻塞性肺病患者外周血嗜酸性粒细胞计数与FeNO水平呈正相关,二者能够反映稳定期慢性阻塞性肺病患者气道的炎症反应,对于稳定期慢性阻塞性肺病患者炎症的评估、病情的程度及后期的治疗具有一定的临床指导意义。     

血清S1P、SFRP1、TIM4、SFRP5与成人支气管哮喘急性发作期患者肺功能、气道炎症和治疗后再次急性复发的关系

摘要 目的：观察血清分泌型卷曲相关蛋白1（SFRP1）、分泌型卷曲相关蛋白5（SFRP5）、1-磷酸鞘氨醇（S1P）、T细胞免疫球蛋白域及黏蛋白域蛋白4（TIM4）与成人支气管哮喘急性发作期患者肺功能、气道炎症和治疗后再次急性复发的关系。方法：选择火箭军特色医学中心2016年7月~2020年8月期间收治的120例成人支气管哮喘患者,其中47例缓解期患者纳为缓解组,73例急性发作期患者纳为发作组。对比发作组、缓解组血清S1P、SFRP1、TIM4、SFRP5水平、肺功能[第1秒用力呼气容积（FEV₁）、用力肺活量（FVC）、FEV₁/FVC]、气道炎症[血清总免疫球蛋白E（IgE）、痰嗜酸粒细胞、呼出气一氧化氮（FeNO）、血嗜酸粒细胞],治疗结束后以门诊复查或电话的形式进行随访1年,根据1年内是否再次急性复发分组,分为复发组和未复发组,对比复发组和未复发组的血清S1P、SFRP1、TIM4、SFRP5水平。结果：发作组的S1P、SFRP1、TIM4高于缓解组,SFRP5低于缓解组（P<0.05）。发作组的FEV₁、FVC、FEV₁/FVC低于缓解组（P<0.05）。发作组的血清总IgE、嗜酸粒细胞、FeNO、血嗜酸粒细胞高于缓解组（P<0.05）。Pearson相关性分析结果显示,S1P、SFRP1、TIM4与FEV₁、FVC、FEV₁/FVC呈负相关（P<0.05）,而与血清总IgE、嗜酸粒细胞、FeNO、血嗜酸粒细胞均呈正相关（P<0.05）。SFRP5与FEV₁、FVC、FEV₁/FVC呈正相关（P<0.05）,而与血清总IgE、嗜酸粒细胞、FeNO、血嗜酸粒细胞均呈负相关（P<0.05）。复发组的S1P、SFRP1、TIM4高于未复发组,SFRP5低于未复发组（P<0.05）。结论：成人支气管哮喘急性发作期患者S1P、SFRP1、TIM4水平异常升高,SFRP5异常降低,且与肺功能、气道炎症、再次急性复发均有一定关系。     

CUL1基因对MPP+诱导的SH-SY5Y细胞存活和NLRP3炎症体通路的影响

摘要 目的：探究Cullin1（CUL1）基因对1-甲基-4-苯基吡啶离子（MPP+）诱导的SH-SY5Y细胞存活和核苷酸结合寡聚化结构域样受体3（NLRP3）炎症体通路的影响。方法：（1）将SH-SY5Y细胞分为NC组、NC-sh组、CUL1-sh组、NC-OE组和CUL1-OE组。使用Lipofectamine 2000试剂对细胞转染相应的慢病毒。（2）将SH-SY5Y细胞分为Control组、MPP+组和MPP++CUL1-OE组。MPP+组和MPP++CUL1-OE组细胞使用1 mmol/L的MPP+处理48 h,Control组细胞正常培养。通过MTT法检测细胞增殖,通过Annexin V-FITC/PI双染色法和TUNEL染色法检测细胞凋亡,通过qRT-PCR检测CUL1的mRNA水平,通过Western blot检测CUL1、NLRP3、凋亡相关斑点样蛋白（ASC）、cleaved caspase-1、白细胞介素（IL）-1β和IL-18蛋白水平。通过ELISA法检测细胞培养上清液中IL-1β和IL-18水平。结果：（1）与NC组和NC-sh组比较,CUL1-sh组CUL1的mRNA和蛋白相对表达量降低,相对细胞活力降低,Annexin V-FITC/PI阳性率和TUNEL阳性率升高,NLRP3、ASC、cleaved caspase-1、IL-1β和IL-18蛋白相对表达量以及细胞培养上清液中IL-1β和IL-18水平升高（P<0.05）。与NC组和NC-OE组比较,CUL1-OE组CUL1的mRNA和蛋白相对表达量升高,相对细胞活力升高,Annexin V-FITC/PI阳性率和TUNEL阳性率降低,NLRP3、ASC、cleaved caspase-1、IL-1β和IL-18蛋白相对表达量以及细胞培养上清液中IL-1β和IL-18水平降低（P<0.05）。（2）与Control组比较,MPP+组CUL1的mRNA和蛋白相对表达量降低,相对细胞活力降低,Annexin V-FITC/PI阳性率和TUNEL阳性率升高,NLRP3、ASC、cleaved caspase-1、IL-1β和IL-18蛋白相对表达量以及细胞培养上清液中IL-1β和IL-18水平升高（P<0.05）。与MPP+组比较,MPP++CUL1-OE组CUL1的mRNA和蛋白相对表达量升高,相对细胞活力升高,Annexin V-FITC/PI阳性率和TUNEL阳性率降低,NLRP3、ASC、cleaved caspase-1、IL-1β和IL-18蛋白相对表达量以及细胞培养上清液中IL-1β和IL-18水平降低（P<0.05）。结论：CUL1可能通过抑制NLRP3炎症体激活促进MPP+诱导的SH-SY5Y细胞存活。     

氧化应激下角质形成细胞中YTHDC1升高促进IL-1β表达

摘要 目的：探讨氧化应激下角质形成细胞内m6A甲基化修饰酶YTHDC1异常对促炎因子的调控机制。方法：通过Western blot和qRT-PCR实验检测氧化应激下角质形成细胞中YTHDC1蛋白和mRNA表达水平。siRNA转染至角质形成细胞以干涉YTHDC1表达,随后继续给予300 μM过氧化氢处理,通过Western blot和qRT-PCR实验检测角质形成细胞中促炎因子IL-1β蛋白和mRNA表达,进一步通过ELISA检测细胞上清中IL-1β分泌,通过CCK8法检测细胞存活水平。结果：1）过氧化氢刺激后人角质形成细胞系HaCaT细胞中YTHDC1表达水平较未处理组明显升高;2）干涉YTHDC1可以显著降低HaCaT细胞中IL-1β表达和上清中分泌;3）干涉YTHDC1后IL-1β mRNA稳定性下降,并且细胞存活率下降。结论：氧化应激下角质形成细胞中m6A甲基化修饰酶YTHDC1表达水平升高,通过提高mRNA稳定性促进IL-1β表达,可能是外界环境应激引起各种免疫性皮肤病的重要机制。     

诱导痰黏蛋白5AC对支气管哮喘和慢性阻塞性肺疾病的鉴别价值

摘要 目的：对支气管哮喘（BA）患者和慢性阻塞性肺疾病（COPD）患者诱导痰黏蛋白5AC（MUC5AC）、炎症细胞和炎症因子水平进行比较以及相关性分析,评估MUC5AC鉴别BA和COPD的价值。方法：选取2018年9月至2019年9月来海南医学院第一附属医院就诊的BA稳定期患者（BA组）60例,同期COPD稳定期（COPD组）患者60例。诱导痰法采样并处理痰液,检测诱导痰MUC5AC、炎症细胞中性粒细胞（Neu）、巨噬细胞（Mac）、嗜酸性粒细胞（Eos）、淋巴细胞（Lym）、炎症因子血管内皮生长因子（VEGF）、细胞间黏附分子-1（ICAM-1）、白介素13（IL-13）和白介素17（IL-17）水平,通过Pearson相关分析对MUC5AC水平与Neu、Mac、Eos、Lym、VEGF、ICAM-1、IL-13、IL-17的关系进行分析。此外,采用受试者工作特征(ROC)曲线分析MUC5AC、炎症细胞及炎症因子鉴别诊断BA及COPD的效能。结果：COPD组诱导痰MUC5AC水平高于BA组,差异有统计学意义（P＜0.05）;COPD组诱导痰Mac和Eos水平均低于BA组,COPD组诱导痰Neu水平高于BA组,差异均有统计学意义（P＜0.05）;COPD组诱导痰VEGF、ICAM-1、IL-13和IL-17水平均低于BA组,差异均有统计学意义（P＜0.05）;Pearson相关分析结果显示,诱导痰MUC5AC与炎症细胞Mac、Eos和炎症因子VEGF、ICAM-1、IL-13和IL-17呈负相关（P＜0.05）,与炎症细胞Neu呈正相关（P＜0.05）。经ROC曲线分析可得：诱导痰MUC5AC鉴别BA和COPD的曲线下面积、灵敏度、特异度以及约登指数均高于炎症细胞Neu、Mac、Eos以及炎症因子VEGF、ICAM-1、IL-13、IL-17。结论：COPD患者诱导痰MUC5AC水平高于BA患者,MUC5AC与炎症细胞和炎症因子有关,MUC5AC的检测有助于鉴别BA或COPD,其有望作为临床BA或COPD的监测指标和治疗靶点。     

miR-29a对于膝关节骨性关节炎大鼠滑膜损伤中的保护作用研究

摘要 目的：探讨miR-29a对于膝关节骨性关节炎（KOA）大鼠滑膜损伤中的保护作用研究。方法：采用前交叉韧带横断法（ACLT）建立KOA大鼠模型。大鼠注射microRNA阴性对照和miR-29a。通过实时定量聚合酶链反应（RT-qPCR）检测KOA滑膜组织和滑膜细胞中miR-29a的表达。RT-qPCR和蛋白免疫印迹试验检测Toll样受体4/髓样分化蛋白88/核因子κB（TLR4/Myd88/NF-κB）信号通路相关蛋白的表达。检测KOA滑膜组织及滑膜细胞中炎症因子的表达水平。结果：KOA滑膜组织和滑膜细胞中miR-29a表达下调。上调miR-29a可抑制KOA大鼠滑膜细胞的炎症反应,促使KOA大鼠的TLR4/Myd88/NF-κB信号通路失活。结论：上调miR-29a可通过TLR4/Myd88/NF-κB信号通路失活化抑制KOA大鼠滑膜细胞炎症反应,从而保护滑膜损伤。     

嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值分析

摘要 目的：探讨嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值。方法：选取我院2020年3月到2023年3月收治的100例慢性鼻窦炎伴鼻息肉患者作为研究对象,将其分为慢性鼻窦炎伴鼻息肉组,选取同期来我院治疗的100例单纯慢性鼻窦炎患者作为慢性鼻窦炎组,另选取同期来我院体检的100名健康志愿者作为对照组。对比三组受检者嗜酸性粒细胞、调节性T细胞表达水平,并建立受试者工作特征（ROC）曲线,分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断效能。随后对100例性鼻窦炎伴鼻息肉患者手术治疗后进行1年随访,依照患者的复发情况评价其预后情况,并分为两个亚组,将术后1年内复发的21例患者分为预后不良组,两未复发的79例患者分为预后良好组,对比两组患者一般临床情况,嗜酸性粒细胞、调节性T细胞表达水平,并分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的预后预测价值。结果：三组受检者嗜酸性粒细胞、调节性T细胞表达水平对比差异显著,慢性鼻窦炎伴鼻息肉组患者嗜酸性粒细胞（EOS）个数高于慢性鼻窦炎组和对照组,调节性T细胞（Tregs）水平低于慢性鼻窦炎组和对照组（P＜0.05）;通过绘制ROC曲线,确定EOS、Tregs其对慢性鼻窦炎伴鼻息肉的诊断效能,结果显示,EOS、Tregs两者联合对慢性鼻窦炎伴鼻息肉的诊断效能优于单一检测（P＜0.05）;预后良好组与预后不良组患者性别、年龄、BMI、吸烟史、饮酒史、白细胞介素-35（IL-35）、转化生长因子-β（TGF-β）、白细胞介素-1β（IL-1β）表达水平对比无明显差异（P＞0.05）,预后良好组与预后不良组患者病程、合并哮喘、合并变应性鼻炎、组织淋巴细胞占比、白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）、EOS个数以及Tregs表达水平对比差异显著（P＜0.05）;logistic回归分析结果表明：合并哮喘、组织淋巴细胞占比、EOS个数以及Tregs为慢性鼻窦炎伴鼻息肉的预后独立影响因素（P＜0.05）。结论：嗜酸性粒细胞、调节性T细胞不仅对慢性鼻窦炎伴鼻息肉的临床诊断具有重要价值,而且能够预测慢性鼻窦炎伴鼻息肉患者的预后情况,因此临床上对于EOS个数增加,Tregs降低的患者要及时改善治疗措施,预防患者预后不良的发生。     

SOX2影响PD-L1表达参与乳腺癌细胞免疫逃逸的机制探讨

摘要 目的：探讨转录因子性别决定区Y框蛋白2（SOX2）基因通过诱导程序性死亡因子配体1（PD-L1）表达,促进乳腺癌细胞免疫逃逸的作用机制。方法：qRT-PCR和Western blot法检测人乳腺癌细胞系MDA-MB-231、SK-BR-3和正常乳腺上皮细胞系MCF10A中SOX2、PD-L1、T细胞免疫球蛋白及粘蛋白结构域分子3（TIM3）和核受体亚家族4 A组成员1（NR4A1）的mRNA表达量及对应蛋白表达量。分别构建SOX2敲减质粒NC和si-SOX2并转染MDA-MB-231,与细胞因子诱导的杀伤细胞构建共培养体系,连续培养48h,MTT法检测细胞增殖率,TUNEL法检测细胞凋亡率,乳酸脱氢酶（LDH）释放实验检测 T细胞杀伤率,ELISA检测T细胞活化标志物IL-2和IFN-γ,qRT-PCR 检测 T 细胞耗竭标志物 PD-1、TIM3和NR4A1 mRNA表达量。结果：与MCF10A相比,MDA-MB-231与SK-BR-3中SOX2、PD-L1、TIM3、NR4A1的mRNA表达量及对应蛋白表达量均显著升高（P＜0.05）。与空白组和NC组相比,si-SOX2组SOX2 mRNA和PD-L1蛋白表达量明显下降,细胞增殖率显著下降,细胞凋亡率增加,T细胞杀伤率增加,IL-2和IFN-γ水平升高,PD-1、TIM3和NR4A1 mRNA表达量降低（P＜0.05）。结论：SOX2基因可能通过诱导PD-L1高表达,进而促使乳腺癌细胞免疫逃逸的能力增强,沉默SOX2表达可以增强T细胞杀伤力,抑制肿瘤增殖,SOX2基因有望成为乳腺癌干预的重要分子靶点。     

The Induction of IL-33 in the Sinus Epithelium and Its Influence on T-Helper Cell Responses

BackgroundChronic rhinosinusitis (CRS) is characterized by epithelial activation and chronic T-cell infiltration in sinonasal mucosa and nasal polyps. IL-33 is a new cytokine of the IL-1 cytokine family that has a pro-inflammatory and Th2 type cytokine induction property. The role of IL-33 in the pathomechanisms of CRS and its interaction with other T cell subsets remain to be fully understood.MethodsThe main trigger for IL-33 mRNA expression in primary human sinonasal epithelial cells was determined in multiple cytokine and T-cell stimulated cultures. The effects of IL-33 on naïve, Th0 and memory T-cells was studied by PCR, ELISA and flow cytometry. Biopsies from sinus tissue were analyzed by PCR and immunofluorescence for the presence of different cytokines and receptors with a special focus on IL-33.ResultsIL-33 was mainly induced by IFN-γ in primary sinonasal epithelial cells, and induced a typical CRSwNP Th2 favoring cytokine profile upon co-culture with T-helper cell subsets. IL-33 and its receptor ST2 were highly expressed in the inflamed epithelial tissue of CRS patients. While IL-33 was significantly up-regulated in the epithelium for CRSsNP, its receptor was higher expressed in sinus tissue from CRSwNP.ConclusionsThe present study delineates the influence of IL-33 in upper airway epithelium and a potential role of IL-33 in chronic inflammation of CRSwNP by enhancing Th2 type cytokine production, which could both contribute to a further increase of an established Th2 profile in CRSwNP.     

Increased expression of chemokine receptors CCR1 and CCR3 in nasal polyps: molecular basis for recruitment of the granulocyte infiltrate

Inflammatory processes play an important role in the development of nasal polyps (NP), but the etiology and, to a high degree also, the pathogenesis of NP are not fully understood. The role of several cytokines and chemokines such as eotaxins, IL-4, IL-5, IL-6, IL-8, and RANTES has been reported in NP. Herewith, we investigated the expression and pattern of distribution of chemokine receptors CCR1 and CCR3 in nasal polyps. Immunohistochemical detection was carried out in frozen sections of biopsies from 22 NP and 18 nasal mucosa specimens in both the epithelial and stromal compartments. Fluorescence microscopy and computerized image analysis revealed a statistically significant increased number of CCR1 (45.2?±?2.8 vs. 15.1?±?1.9, p?<?0.001)-positive as well as CCR3 (16.4?±?1.4 vs. 9.7?±?1.1, p?<?0.001)-positive cells in the stroma of NP compared to nasal mucosa. In comparison to healthy nasal mucosa, increased positivity of CCR3 was detected in the epithelial compartment of NP. Our data suggest that increased expression of CCR1 and CCR3 chemokine receptors may, in accord with various chemokines, contribute to the pathogenesis of nasal polyposis by facilitating increased migration and prolonged accumulation of inflammatory cells, e.g., eosinophils, in the inflammatory infiltrate of NP.     

趋化因子CCL-18 在慢性鼻- 鼻窦炎中的表达及意义

目的:分析趋化因子CCL-18在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜的表达差异,探讨CCL-18在慢性鼻-鼻窦炎中的表达及意义。方法:采用苏木精-伊红染色(HE),Masson染色及过碘酸-雪夫(PAS)染色对慢性鼻-鼻窦炎组织进行病理分析。采用Western blot检测CCL-18蛋白水平在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜组织中的表达差异。结果:CCL-18蛋白水平在伴鼻息肉和不伴有鼻息肉慢性鼻窦炎均较正常鼻黏膜组织中显著上调(P<0.05)。CCL-18蛋白水平在嗜酸性粒细胞慢性鼻窦炎的表达水平明显高于非嗜酸性粒细胞慢性鼻窦炎(P<0.05)。腺体型,纤维炎症型及水肿型慢性鼻-鼻窦炎中CCL-18表达水平均高于正常鼻黏膜,以水肿型表达最为显著(P<0.05)。结论:CCL-18在嗜酸性粒细胞和水肿型慢性鼻窦炎中高度表达,提示CCL-18可能参与慢性鼻-鼻窦炎中嗜酸性粒细胞的浸润这一基本病理过程。     

Deprived TLR9 Expression in Apparently Healthy Nasal Mucosa Might Trigger Polyp-Growth in Chronic Rhinosinusitis Patients

Background

The origin of nasal polyps in chronic rhinosinusitis is unknown, but the role of viral infections in polyp growth is clinically well established. Toll-like receptors (TLRs) have recently emerged as key players in our local airway defense against microbes. Among these, TLR9 has gained special interest in viral diseases. Many studies on chronic rhinosinusitis with nasal polyps (CRSwNP) compare polyp tissue with nasal mucosa from polyp-free individuals. Knowledge about changes in the turbinate tissue bordering the polyp tissue is limited.

Objectives

To analyse the role of TLR9 mediated microbial defense in tissue bordering the polyp.

Methods

Nasal polyps and turbinate tissue from 11 patients with CRSwNP and turbinate tissue from 11 healthy controls in total were used. Five biopsies from either group were analysed immediately with flow cytometry regarding receptor expression and 6 biopsies were used for in vitro stimulation with a TLR9 agonist, CpG. Cytokine release was analysed using Luminex. Eight patients with CRSwNP in total were intranasally challenged with CpG/placebo 24 hours before surgery and the biopsies were collected and analysed as above.

Results

TLR9 expression was detected on turbinate epithelial cells from healthy controls and polyp epithelial cells from patients, whereas TLR9 was absent in turbinate epithelial cells from patients. CpG stimulation increased the percentage cells expressing TLR9 and decreased percentage cells expressing VEGFR2 in turbinate tissue from patients. After CpG stimulation the elevated levels of IL-6, G-CSF and MIP-1β in the turbinate tissue from patients were reduced towards the levels demonstrated in healthy controls.

Conclusion

Defects in the TLR9 mediated microbial defense in the mucosa adjacent to the anatomic origin of the polyp might explain virus induced polyp growth. CpG stimulation decreased VEGFR2, suggesting a role for CpG in polyp formation. The focus on turbinate tissue in patients with CRSwNP opens new perspectives in CRSwNP-research.     

Interleukin-13 Inhibits Lipopolysaccharide-Induced BPIFA1 Expression in Nasal Epithelial Cells

Short palate, lung, and nasal epithelium clone 1 (SPLUNC1) protein is expressed in human nasopharyngeal and respiratory epithelium and has demonstrated antimicrobial activity. SPLUNC1 is now referred to as bactericidal/permeability-increasing fold containing family A, member 1 (BPIFA1). Reduced BPIFA1 expression is associated with bacterial colonization in patients with chronic rhinosinusitis with nasal polyps (CRSwNP). Interleukin 13 (IL-13), predominately secreted by T helper 2 (T_H2) cells, has been found to contribute to airway allergies and suppress BPIFA1 expression in nasal epithelial cells. However, the molecular mechanism of IL-13 perturbation of bacterial infection and BPIFA1 expression in host airways remains unclear. In this study, we found that lipopolysaccharide (LPS)-induced BPIFA1 expression in nasal epithelial cells was mediated through the JNK/c-Jun signaling pathway and AP-1 activation. We further demonstrated that IL-13 downregulated the LPS-induced activation of phosphorylated JNK and c-Jun, followed by attenuation of BPIFA1 expression. Moreover, the immunohistochemical analysis showed that IL-13 prominently suppressed BPIFA1 expression in eosinophilic CRSwNP patients with bacterial infection. Taken together, these results suggest that IL-13 plays a critical role in attenuation of bacteria-induced BPIFA1 expression that may result in eosinophilic CRSwNP.     

Prostaglandin E2 receptors in asthma and in chronic rhinosinusitis/nasal polyps with and without aspirin hypersensitivity

Chronic rhinosinusitis with nasal polyps (CRSwNP) and asthma frequently coexist and are always present in patients with aspirin exacerbated respiratory disease (AERD). Although the pathogenic mechanisms of this condition are still unknown, AERD may be due, at least in part, to an imbalance in eicosanoid metabolism (increased production of cysteinyl leukotrienes (CysLTs) and reduced biosynthesis of prostaglandin (PG) E₂), possibly increasing and perpetuating the process of inflammation. PGE₂ results from the metabolism of arachidonic acid (AA) by cyclooxygenase (COX) enzymes, and seems to play a central role in homeostasis maintenance and inflammatory response modulation in airways. Therefore, the abnormal regulation of PGE₂ could contribute to the exacerbated processes observed in AERD. PGE₂ exerts its actions through four G-protein-coupled receptors designated E-prostanoid (EP) receptors EP1, EP2, EP3, and EP4. Altered PGE₂ production as well as differential EP receptor expression has been reported in both upper and lower airways of patients with AERD. Since the heterogeneity of these receptors is the key for the multiple biological effects of PGE₂ this review focuses on the studies available to elucidate the importance of these receptors in inflammatory airway diseases.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-014-0100-7) contains supplementary material, which is available to authorized users.     

Effect of Lipopolysaccharide on Glucocorticoid Receptor Function in Control Nasal Mucosa Fibroblasts and in Fibroblasts from Patients with Chronic Rhinosinusitis with Nasal Polyps and Asthma

BackgroundChronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammatory disease of the upper airways frequently associated with asthma. Bacterial infection is a feature of CRSwNP that can aggravate the disease and the response to glucocorticoid treatment.ObjectiveWe examined whether the bacterial product lipopolysaccharide (LPS) reduces glucocorticoid receptor (GR) function in control nasal mucosa (NM) fibroblasts and in nasal polyp (NP) fibroblasts from patients with CRSwNP and asthma.MethodsNP (n = 12) and NM fibroblasts (n = 10) were in vitro pre-incubated with LPS (24 hours) prior to the addition of dexamethasone. Cytokine/chemokine secretion was measured by ELISA and Cytometric Bead Array. GRα, GRβ, mitogen-activated protein-kinase phosphatase-1 (MKP-1) and glucocorticoid-induced leucine zipper (GILZ) expression was measured by RT-PCR and immunoblotting, GRα nuclear translocation by immunocytochemistry, and GRβ localization by immunoblotting. The role of MKP-1 and GILZ on dexamethasone-mediated cytokine inhibition was analyzed by small interfering RNA silencing.ResultsPre-incubation of nasal fibroblasts with LPS enhanced the secretion of IL-6, CXCL8, RANTES, and GM-CSF induced by FBS. FBS-induced CXCL8 secretion was higher in NP than in NM fibroblasts. LPS effects on IL-6 and CXCL8 were mediated via activation of p38α/β MAPK and IKK/NF-κB pathways. Additionally, LPS pre-incubation: 1) reduced dexamethasone’s capacity to inhibit FBS-induced IL-6, CXCL8 and RANTES, 2) reduced dexamethasone-induced GRα nuclear translocation (only in NM fibroblasts), 3) did not alter GRα/GRβ expression, 4) decreased GILZ expression, and 5) did not affect dexamethasone’s capacity to induce MKP-1 and GILZ expression. MKP-1 knockdown reduced dexamethasone’s capacity to suppress FBS-induced CXCL8 release.ConclusionThe bacterial product LPS negatively affects GR function in control NM and NP fibroblasts by interfering with the capacity of the activated receptor to inhibit the production of pro-inflammatory mediators. This study contributes to the understanding of how bacterial infection of the upper airways may limit the efficacy of glucocorticoid treatment.     

Spontaneous eosinophilic nasal inflammation in a genetically-mutant mouse: comparative study with an allergic inflammation model

Background

Eosinophilic inflammation is a hallmark of chronic rhinosinusitis with nasal polyps. To model this disease process experimentally, nasal sensitization of mice with ovalbumin or aspergillus has been described. Here, we describe a genetically mutant mouse that develops robust spontaneous nasal eosinophilic inflammation. These mice lack the enzyme SHP-1 that down-regulates the IL-4Rα/stat6 signaling pathway. We compared nasal inflammation and inflammatory mediators in SHP-1 deficient mice (mev) and an ovalbumin-induced nasal allergy model.

Methods

A novel technique of trans-pharyngeal nasal lavage was developed to obtain samples of inflammatory cells from the nasal passages of allergic and mev mice. Total and differential cell counts were performed on cytospin preparations. Expression of tissue mRNA for IL-4, IL-13, and mouse beta-defensin-1 (MBD-1) was determined by quantitative PCR. Eotaxin in the lavage fluid was assessed by ELISA.

Results

Allergic and mev mice had increased total cells and eosinophils compared with controls. Expression of IL-4 was similarly increased in both allergic and mev mice, but expression of IL-13 and eotaxin was significantly greater in the allergic mice than mev mice. Eotaxin was significantly up-regulated in both allergic rhinitis and mev mice. In both models of eosinophilic inflammation, down-regulation of the innate immune marker MBD-1 was observed.

Conclusions

The mev mice display spontaneous chronic nasal eosinophilic inflammation with potential utility for chronic rhinosinusitis with nasal polyps research. The eosinophilic infiltrate is more robust in the mev mice than allergic mice, but Th2 cytokine expression is not as pronounced. Decreased MBD-1 expression in both models supports the concept that Th2-cytokines down-regulate sinonasal innate immunity in humans, and suggests a role for mouse models in investigating the interaction between adaptive and innate immunity in the sinonasal mucosa.