藏红花素在嗜酸性慢性鼻窦炎伴鼻息肉中调控2型炎症反应的研究  

Effect of Crocin on Type 2 inflammation in eosinophilic chronic rhinosinusitis with nasal polyps

作　　者：张婕 尼玛吉宗[2] 徐小东 周菁 刘建敏 罗依蕤 杜进涛[4] 巴罗 ZHANG Jie;NIMA Jizong;XU Xiaodong;ZHOU Jing;LIU Jianmin;LUO Yirui;DU Jintao;BA Luo(Medical College of Tibet University,Lasa 850000,Xizang,China;Department of Otorhinolaryngology,People's Hospital of Tibet Autonomous Region,Lasa 850000,Xizang,China;Department of Otolaryngology,Shannan City People's Hospital,Shannan 856000,Xizang,China;Department of Otorhinolaryngology&Head and Neck Surgery,West China Hospital,Sichuan University,Chengdu 610041,Sichuan,China)

机构地区：[1]西藏大学医学院,西藏拉萨850000 [2]西藏自治区人民医院耳鼻咽喉科,西藏拉萨850000 [3]山南市人民医院耳鼻喉科,西藏山南856000 [4]四川大学华西医院耳鼻咽喉头颈外科,四川成都610000

出　　处：《山东大学耳鼻喉眼学报》2025年第1期61-67,104,共8页Journal of Otolaryngology and Ophthalmology of Shandong University

基　　金：国家自然科学基金(81860186);西藏自治区中央引导地方科技发展资金项目(ZYYD2023000133)。

摘　　要：目的探讨藏红花素在嗜酸性粒细胞性慢性鼻窦炎伴鼻息肉(eosinophilic chronic rhinosinusitis with nasal polyps,Eos CRSwNP)患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)2型炎症反应中的调控作用和作用机制,为精准治疗鼻息肉提供新的靶点或通路。方法收集慢性鼻窦炎伴鼻息肉(chronic rhinosinusitis with nasal polyps,CRSwNP)患者鼻息肉组织进行病理学诊断,根据炎症细胞类型分为嗜酸性粒细胞性慢性鼻窦炎伴鼻息肉(eosinophilic chronic rhinosinusitis with nasal polyps,Eos CRSwNP)和非嗜酸性粒细胞性慢性鼻窦炎伴鼻息肉(non-eosinophilic chronic rhinosinusitis with nasal polyps,Non-Eos CRSwNP)。密度梯度离心法提取Eos CRSwNP患者外周血中的PBMC。用不同浓度IL-33刺激PBMC,qPCR检测转录因子GATA3、T-bet和细胞受体CRTH2,qPCR和ELISA分析炎症细胞因子INF-γ,IL-5和IL-13的表达情况。再加入不同浓度藏红花素进行干预,Western blotting和qPCR检测GATA3、T-bet和CRTH2。Graphpad prism软件进行统计分析,对比IL-33刺激前后、藏红花素干预前后指标变化有无显著差异。结果共有16例经病理学确诊为Eos CRSwNP。不同浓度的IL-33刺激PBMC后,qPCR证实T-bet,GATA3和CRTH2的表达有明显升高(P值分别为0.004、0.001、0.026);qPCR和ELISA进一步发现INF-γ,IL-5和IL-13等细胞因子上调(qPCR的P值分别为0.001、0.001、0.003;ELISA的P值分别为0.035、0.006、0.001),其中IL-33浓度为20 ng/mL时对PBMC炎症细胞活化最为明显。再加入藏红花素干预后,Western blotting和qPCR检测显示T-bet,GATA3和CRTH2表达呈下降趋势,且在浓度为10 ug/mL时最为明显(WB的P值分别为0.027、0.001、0.002;qPCR的P值分别为0.019、0.001、0.001)。结论藏红花素能够显著抑制IL-33激活的CRSwNP患者内的2型炎症反应,具有潜在的CRSwNP新型治疗药物开发价值。Objective To study the effect of crocin on Eosinophilic Chronic Rhinosinusitis with Nasal Polyps,the regulatory role and mechanism of Peripheral blood mononuclear cell(PBMC)type 2 inflammation in patients with Eos CRSwNP,providing a new target or pathway for precise treatment of nasal polyps.Methods Chronic rhinosinusitis with nasal polyps(CRSwNP)patients were collected for pathological diagnosis.Eosinophilic Chronic Rhinosinusitis with Nasal Polyps(Eos CRSwNP)and Non-eosinophilic Chronic Rhinosinusitis with Nasal Polyps(Non-Eos CRSwNP)were classified according to the inflammatory cell types.PBMC in peripheral blood of Eos CRSwNP patients was extracted by density gradient centrifugation.PBMC was stimulated with different concentrations of IL-33,and transcription factors GATA3,T-bet,and cell receptor CRTH2 were detected by qPCR.The expression of inflammatory cytokines INF-γ,IL-5,and IL-13 was analyzed by qPCR and ELISA.Then different concentrations of crocin were added for intervention,and GATA3,T-bet,and CRTH2 were detected by Western blotting and qPCR.GraphPad Prism software was used for statistical analysis to compare whether there were significant differences in index changes before and after IL-33 stimulation and crocin intervention.Results A total of 16 cases were pathologically confirmed as Eos CRSwNP.After stimulating PBMC with different concentrations of IL-33,qPCR confirmed that the expression of T-bet,GATA3,and CRTH2 was significantly increased(P values were 0.004,0.001,and 0.026,respectively).qPCR and ELISA further revealed upregulation of cytokines INF-γ,IL-5,and IL-13(P values of qPCR were 0.001,0.001,and 0.003,respectively;P values of ELISA were 0.035,0.006,and 0.001,respectively),with the most significant activation of PBMC inflammatory cells observed at an IL-33 concentration of 20 ng/mL.After intervention with crocin,Western blotting and qPCR showed a downward trend in the expression of T-bet,GATA3,and CRTH2,with the most significant effect observed at a concentration of 10μg/mL(P values of WB

关 键 词：慢性鼻窦炎 鼻息肉 嗜酸性粒细胞 外周血单个核细胞 藏红花素 2型炎症反应 

分 类 号：R765.41[医药卫生—耳鼻咽喉科] R392.11[医药卫生—临床医学]