METHODS: Quantitative PCR, immunofluorescence staining, western blotting, and enzyme-linked immunosorbent assay (ELISA) were used to analyze the expression of sPLA2s, phospholipase A2 receptor (PLA2R), and prostaglandin D2 (PGD2) in nasal samples. Human nasal epithelial cells (HNECs) were cultured at an air-liquid interface (ALI) and stimulated with various cytokines. The human mast cell line HMC-1 was stimulated with sPLA2GIB, and the expression of PGD2 and cytokines in the culture supernatant was detected by ELISA.
RESULTS: The mRNA and protein levels of sPLA2GIB were significantly higher in eosinophilic CRSwNP than in control tissues. sPLA2GIB was predominantly expressed in the nasal epithelial cells. PLA2R mRNA and protein levels were upregulated in both eosinophilic and non-eosinophilic CRSwNP compared with the control groups. IL-4, IL-13, TNF-α, and IL-1β upregulated the expression of sPLA2GIB in ALI-cultured HNECs. sPLA2GIB induced PGD2 and IL-13 production in HMC-1 cells in a hydrolytic activity-independent manner. PGD2 protein expression was elevated in tissue homogenates of eosinophilic CRSwNP, and PGD2 upregulated the expression of IL-13 in HMC-1 cells.
CONCLUSIONS: Increased secretion of sPLA2GIB by epithelial cells may promote eosinophilic inflammation in CRSwNP by enhancing PGD2 and IL-13 production in mast cells via binding to PLA2R.
METHODS: N/A Laryngoscope, 134:1107-1117, 2024.
方法:定量PCR,免疫荧光染色,西方印迹,酶联免疫吸附试验(ELISA)分析sPLA2s的表达,磷脂酶A2受体(PLA2R),和鼻部样本中的前列腺素D2(PGD2)。在气液界面(ALI)培养人鼻上皮细胞(HNEC),并用各种细胞因子刺激。用sPLA2GIB刺激人肥大细胞系HMC-1,ELISA法检测培养上清液中PGD2和细胞因子的表达。
结果:嗜酸性粒细胞CRSwNP中sPLA2GIB的mRNA和蛋白水平明显高于对照组织。sPLA2GIB主要在鼻上皮细胞中表达。与对照组相比,嗜酸性粒细胞和非嗜酸性粒细胞CRSwNP中的PLA2RmRNA和蛋白水平均上调。IL-4,IL-13,TNF-α,IL-1β上调ALI培养的HNECs中sPLA2GIB的表达。sPLA2GIB以不依赖水解活性的方式诱导HMC-1细胞中PGD2和IL-13的产生。嗜酸性粒细胞CRSwNP组织匀浆中PGD2蛋白表达升高,PGD2上调HMC-1细胞中IL-13的表达。
结论:上皮细胞分泌sPLA2GIB的增加可能通过与PLA2R结合而增强肥大细胞中PGD2和IL-13的产生,从而促进CRSwNP中嗜酸性粒细胞炎症。
方法:N/A喉镜,2023年。