Ruan Jian-Wen, Zhao Jie-Fang, Li Xue-Li, Liao Bo, Pan Li, Zhu Ke-Zhang, Feng Qi-Miao, Liu Jin-Xin, Yu Zi-E, Song Jia, Wang Hai, Liu Zheng
Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Front Cell Dev Biol. 2021 Dec 17;9:793073. doi: 10.3389/fcell.2021.793073. eCollection 2021.
The mechanisms underlying neutrophilic inflammation in chronic rhinosinusitis with nasal polyps (CRSwNP) remain poorly investigated. This study aimed to examine the factors that contribute to tissue neutrophilia in CRSwNP. The numbers of neutrophils and active caspase-3-positive apoptotic neutrophils in sinonasal tissues were assessed via immunofluorescence staining. The 95th percentile of tissue neutrophil numbers in control subjects was selected as a cut-off to define neutrophil-high (Neu-high) or neutrophil-low (Neu-low) nasal polyps (NPs). The levels of 34 inflammatory mediators in sinonasal tissues were analyzed using Bio-Plex assay. Purified human peripheral blood neutrophils were incubated with nasal tissue homogenates, and the apoptotic neutrophils were assessed flow cytometry. The cut-off for Neu-high NPs was >10 myeloperoxidase positive cells/high-power field. Compared with Neu-low NPs, Neu-high NPs had higher tissue levels of IL-1β, IL-1Ra, IL-6, IL-8, G-CSF, MCP-1, and MIP-1α, but lower levels of IL-5, IL-13, IgE, and eosinophils. Principal component and multiple correspondence analyses revealed mixed type 1, type 2, and type 3 endotypes for Neu-low NPs, and predominant type 1 and type 3 endotypes for Neu-high NPs. Neu-high NPs had lower percentages of apoptotic neutrophils than Neu-low NPs. The numbers of neutrophils and the percentages of apoptotic neutrophils correlated with G-CSF and IL-6 levels in the NPs. Tissue homogenates from Neu-high NPs, but not those from Neu-low NPs, suppressed neutrophil apoptosis , which was reversed by anti-G-CSF treatment. Tissue neutrophil numbers were associated with difficult-to-treat disease in patients with CRSwNP after surgery. We propose that G-CSF promotes neutrophilic inflammation by inhibiting neutrophil apoptosis in CRSwNP.
伴有鼻息肉的慢性鼻-鼻窦炎（CRSwNP）中嗜中性粒细胞炎症的潜在机制仍未得到充分研究。本研究旨在探究导致CRSwNP组织中嗜中性粒细胞增多的因素。通过免疫荧光染色评估鼻窦组织中嗜中性粒细胞和活性半胱天冬酶-3阳性凋亡嗜中性粒细胞的数量。选择对照组中组织嗜中性粒细胞数量的第95百分位数作为临界值，以定义嗜中性粒细胞高（Neu高）或嗜中性粒细胞低（Neu低）的鼻息肉（NP）。使用生物芯片分析（Bio-Plex assay）分析鼻窦组织中34种炎症介质的水平。将纯化的人外周血嗜中性粒细胞与鼻组织匀浆孵育，并通过流式细胞术评估凋亡嗜中性粒细胞。Neu高NP的临界值为>10个髓过氧化物酶阳性细胞/高倍视野。与Neu低NP相比，Neu高NP的组织中IL-1β、IL-1Ra、IL-6、IL-8、G-CSF、MCP-1和MIP-1α水平较高，但IL-5、IL-13、IgE和嗜酸性粒细胞水平较低。主成分分析和多重对应分析显示，Neu低NP为1型、2型和3型内型混合，而Neu高NP主要为1型和3型内型。Neu高NP的凋亡嗜中性粒细胞百分比低于Neu低NP。NP中嗜中性粒细胞数量和凋亡嗜中性粒细胞百分比与G-CSF和IL-6水平相关。Neu高NP的组织匀浆而非Neu低NP的组织匀浆可抑制嗜中性粒细胞凋亡，抗G-CSF治疗可逆转这种抑制作用。CRSwNP患者术后组织嗜中性粒细胞数量与难治性疾病相关。我们提出，G-CSF通过抑制CRSwNP中的嗜中性粒细胞凋亡来促进嗜中性粒细胞炎症。
