Affiliation:
1. Department of Otolaryngology, Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiao
Tong University, Shanghai, 200127, China
Abstract
Aims::
We aim to explore GATA6 modulation in allergic rhinitis (AR).
Background::
Circular RNAs (circRNAs) and microRNAs (miRNAs) are involved in inflammatory
responses; GATA6 is also known to regulate multiple inflammatory pathways.
However, the mechanism of regulation of AR between them is unclear.
Objective::
We expect that this study will provide new treatment options for AR from a
GATA6 perspective.
Methods::
In vitro, AR models were employed to examine the efficacy of our study,
where we utilized monoclonal anti-2,4,6-dinitrophenyl immunoglobulin (Ig) E/human
serum albumin (DNP-IgE/HSA) to induce rat basophilic leukemia cells (RBL-2H3 cells).
Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was performed
to measure the expression of circ_0008668, miR-1301-3p, GATA6, and cellular
inflammatory markers. Enzyme-linked immunosorbent assay (ELISA) was used to measure
concentrations of beta-hexosaminidase, histamine, and cellular inflammatory factors
including TNF-α, IL-1β, IL-4, and IL-5. In addition, western blot, RNA pull-down,
and luciferase assays were performed to validate the molecular mechanism by which circ_
0008668 and miR-1301-3p interactions promote GATA6 to ameliorate the inflammatory
state of RBL-2H3 cells.
Results::
In the in vitro model of AR, the expression levels of circ_0008668 and GATA6
were elevated, whereas that of miR-1301-3p was decreased. Pull-down assays confirmed
that circ_0008668 efficiently binds miR-1301-3p and its overexpression leads to
upregulation of the levels of GATA6, cellular inflammatory factors (IL-4, IL-5, TNF-α,
and IL-1β), and markers associated with inflammatory signaling pathways (NLRP3,
ERK1/2, and P65 protein phosphorylation). In addition, miR-inhibitor with circRNA enhanced
GATA6 and NLPR3 expression and activated inflammatory pathway activity. In
particular, miR-mimic was effective in reversing the onset of this inflammatory state.
Conclusion::
Our results indicate that circ_0008668 promotes the inflammatory state of
mast cells by sponging miR-1301-3p to target GATA6, which in turn affects the allergic
response to AR. This process could improve the current diagnosis of AR patients and
clinical treatment.
Publisher
Bentham Science Publishers Ltd.