IL-6 induces NLRP3 inflammasome activation through JAK/STAT3-dependent NOX2 induction in colon epithelial cells
- Authors
- Kim, J-A; Grung, Premesh; Banskota, Suhrid; Jeong, Byeong-Seon; Nam, Tae-gyu
- Issue Date
- Feb-2017
- Publisher
- OXFORD UNIV PRESS
- Citation
- JOURNAL OF CROHNS & COLITIS, v.11, no.suppl_1, pp.S106 - S107
- Indexed
- SCIE
SCOPUS
- Journal Title
- JOURNAL OF CROHNS & COLITIS
- Volume
- 11
- Number
- suppl_1
- Start Page
- S106
- End Page
- S107
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/10188
- DOI
- 10.1093/ecco-jcc/jjx002.183
- ISSN
- 1873-9946
- Abstract
- Background: IL-6, in addition to TNFα, plays an important role in the pathogenesis of inflammatory bowel disease (IBD), which is supported by clinical observations of close correlation between IL-6 production and severity of the disease in IBD patients. IL-6 plays a role in the recruitment process of neutrophils and monocytes to lesion sites, resulting in aggravated and chronic inflammation. Recently, TNFα is shown to prime TLR-independent NLRP3 inflammasome activation. IL-6, however, has not been shown such activity. The present study aims to investigate whether IL-6 induces NLRP3 inflammasome formation, and NADPH oxidase is involved in that process.
Methods: The IL-6-induced adhesion of monocytes (U937 cell line) to colon epithelial cells (HT-29 cell line) was examined by co-culture of HT-29 cells with U937 cells that were already labeled with BCECF-AM (10 μg/mL). After 3 h treatment with IL-6, BCECF fluorescence from adhered cells was measured. To identify signaling pathway, siRNA transfection, RT-PCR and Western blot analyses were performed. Reactive oxygen species (ROS) was measured by lucigenin chemiluminescence assay.
Results: IL-6 significantly increased U937 monocytic cell adhesion to HT-29 colonic epithelial cells, which was accompanied by upregulation of adhesion molecules (ICAM-1 and VCAM-1), NLRP3, caspase-1, and IL-1β. Concurrently, IL-6 significantly increased ROS production in a time-dependent manner, which matched significant induction of NOX2 and its regulatory subunits. The IL-6-induced ROS production and increased expression of NLRP3, caspase-1, and IL-1β were attenuated by pretreatment with NADPH oxidase inhibitors (VAS-2840, DPI and apocyanin), but not by inhibitors against other enzymes, such as cytosolic COX-2 (celecoxib), mitochondria (antimycin A), xanthine oxidase (allopurinol), and iNOS (NAME) in HT-29 cells. Similarly, inhibitors of JAK (tofacitinib) and STAT3 (stattic) suppressed IL-6-induced ROS production, NOX2 induction, and the changes in inflammasome proteins in HT-29 cells.
Conclusions: Taken together, our results suggest that IL-6 induces NLRP3 inflammasome activation through JAK/STAT-dependent NOX2 induction in HT-29 colonic epithelial cells.
- Files in This Item
-
Go to Link
- Appears in
Collections - COLLEGE OF PHARMACY > DEPARTMENT OF PHARMACY > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.