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ORIGINAL ARTICLE
Hai Lin,1,2,5 Zhipeng Li,1 Dong Lin,3 Chunquan Zheng,4 and Weitian Zhang1,5
Abstract—The pathophysiologic mechanisms of human chronic rhinosinusitis with nasal polyps (CRSwNP)
remain unclear. We aimed to elucidate expression and biologic role of NLRP3 inflammasome in CRSwNP.
Immunohistochemistry (IHC) was conducted to assess NLRP3 immunolabeling, real-time polymerase chain
reaction (PCR) was used for IL-9 and NLRP3, and caspase-1 level quantitation in CRSwNP and control
subjects. In addition, enzyme-linked immunosorbent assay (ELISA) was employed for analyzing concen-
trations of IL-1β and IL-18 in the homogenates prepared from tissue specimens. Moreover, human nasal
epithelial cells (HNECs) were used to evaluate the effects of lipopolysaccharide (LPS) and glyburide on
NLRP3 inflammasome signaling pathway. Results showed that NLRP3 and caspase-1 were overexpressed in
CRSwNP, especially in eosinophilic CRSwNP (ECRSwNP). Interestingly, NLRP3 expression had close
correlation to that of caspase-1. Concentrations of IL-1β and IL-18 were elevated. NLRP3 inflammasome
signaling pathway was augmented by LPS but suppressed by glyburide. In conclusion, NLRP3 inflamma-
some signaling pathway played a pro-inflammatory role in the pathogenesis of CRSwNP, especially in
ECRSwNP. NLRP3 inflammasome signaling pathway was augmented by LPS, but suppressed by glyburide.
KEY WORDS: NLRP3; caspase-1; eosinophil; glyburide; chronic rhinosinusitis; nasal polyps.
2045
0360-3997/16/0600-2045/0 # 2016 Springer Science+Business Media New York
2046 Lin, Li, Lin, Zheng, and Zhang
revealed that NLRP3 plays a pivotal role in the chronic analyzed for concentrations of IL-1β and IL-18 by ELISA.
airway inflammatory diseases like asthma and chronic Additionally, the effects of lipopolysaccharide (LPS) and
obstructive pulmonary disease (COPD) by triggering in- glyburide on NLRP3 inflammasome signaling pathway
flammatory cells recruitment and pro-inflammatory cyto- were evaluated on HNECs culture. This study was ap-
kines release [12–16]. However, its role in the pathogenesis proved by the ethical committees of Shanghai Jiao Tong
of CRSwNP remains uncertain. Therefore, we aimed to University Affiliated Sixth People’s Hospital and Fuzhou
elucidate expression and biologic role of NLRP3 inflam- General Hospital, and an informed consent was provided
masome in CRSwNP. from every subject.
ECRSwNP eosinophilic chronic rhinosinusitis with nasal polyps, nECRSwNP non-eosinophilic chronic rhinosinusitis with nasal polyps, VAS visual analog
scale, N/A not applicable, CT computed tomography
Role of NLRP3 Inflammasome in Eosinophilic and Non-eosinophilic Chronic Rhinosinusitis with Nasal Polyps 2047
Fig. 1. Immunolabeling of NLRP3 in control group (a), nECRSwNP group (b), and ECRSwNP group (c).NLRP3 protein levels were significantly over-
expressed in CRSwNP groups compared with control group and were significantly stronger expressed in ECRSwNP group compared with nECRSwNP
group (a–d). Kruskal–Wallis H test was employed for comparison.*P < 0.05.
Fig. 2. NLRP3, caspase-1, IL-1β, and IL-18 levels were elevated in CRSwNP groups compared with control group, and these were also found in ECRSwNP
group compared with nECRSwNP (a–d). Kruskal–Wallis H test was employed for comparison.*P < 0.05.
Role of NLRP3 Inflammasome in Eosinophilic and Non-eosinophilic Chronic Rhinosinusitis with Nasal Polyps 2049
Fig. 3. Significantly increased NLRP3, caspase-1, IL-1β, and IL-18 levels in cultured HNECs was found after incubation with LPS and LPS + Gly (a–e).
Significant decrease of NLRP3, caspase-1, IL-1β, and IL-18 levels in cultured HNECs was found after incubation with LPS + Gly group compared with LPS
group (a–e). Values were shown as mean ± standard deviation. One-way ANOVA test was used for the statistical analysis.*P < 0.05.
2050 Lin, Li, Lin, Zheng, and Zhang
nECRSwNP group (P < 0.05). These findings were con- glyburide. These findings were in line with the changes in
sistent with the changes in the NLRP3 and caspase-1 levels the NLRP3 and caspase-1 levels in HNECs.
in nasal tissues.
caspase-1 levels in nasal tissues.The results indicated that COMPLIANCE WITH ETHICAL STANDARDS
NLRP3 played a pivotal role in promoting epithelial cell
growth and inflammatory cells infiltration in CRSwNP, and This study was approved by the ethical committees of
the effect of NLRP3 is more obvious in ECRSwNP. Shanghai Jiao Tong University Affiliated Sixth People’s
Considerable data indicate that due to the good repre- Hospital and Fuzhou General Hospital, and an informed
sentative of HNECs for human nasal polyps and convenient consent was provided from every subject.
isolation of HNECs from human nasal polyps, they are most
often applied to deeply explore the regulation of targeted
gene in the pathophysiology of CRSwNP [27–29]. In the
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