Abstract
Calcium-activated potassium ion channel-3.1 (KCa3.1) plays a pivotal role in the potassium-calcium exchange involved in atopy. This study aimed to explore the impact of lentiviral-mediated shRNA silencing KCa3.1 on allergic response in a murine allergic rhinitis (AR) model. The BALB/c mice were divided into four groups: untreated AR group, negative control AR group, lentiviral KCa3.1-shRNA treated AR group and normal control group. Concentrations of ovalbumin (OVA)-specific IgE, histamine and leukotrienes C4 (LTC4) in serum, and IL-4, IL-9 and IL-17 in nasal lavage fluid (NLF) were analyzed. Goblet cells and mast cells were counted. KCa3.1 positive cells were counted after immunolabelling by immunofluorescence method. KCa3.1, Mucin 5AC (MUC5AC), and tryptase mRNA levels were determined using real-time polymerase chain reaction. Furthermore, P815 cell line was used to explore the role and mechanism of lentiviral KCa3.1-shRNA on mast cells. The results showed that LV-KCa3.1-shRNA intervention effectively attenuated allergic responses in LV-KCa3.1-shRNA treated mice. LV-KCa3.1-shRNA intervention effectively suppressed KCa3.1 levels and phosphorylation of AKT in P815 cells, leading to the downregulation of tryptase, IL-6 and IL-8 levels. LV-KCa3.1-shRNA intervention effectively attenuated the allergic responses in AR and suppressed mast cell activity by inhibiting PI3K/AKT signaling pathway.
Highlights
RNA interference (RNAi) is a gene-specific RNA degradation process mediated by short interfering RNA and is a promising therapeutic tool directed against a potential therapeutic target in the allergic disorders, infectious diseases and cancers[9,10,11]
LV-KCa3.1-short hairpin RNA (shRNA) treatment resulted in the reduction of OVA-specific immunoglobulin E (IgE), histamine, leukotrienes C4 (LTC4), IL-4, IL-9 and IL-17 levels in LV-KCa3.1-shRNA treated group compared with untreated Allergic rhinitis (AR) or negative control group, respectively (P < 0 .01)
It has been proved that goblet cell hyperplasia, mast cell infiltration, Th-2, Th-9 and Th-17 polarization are recognized as important features of AR3,5,22,23
Summary
RNA interference (RNAi) is a gene-specific RNA degradation process mediated by short interfering RNA (siRNA) and is a promising therapeutic tool directed against a potential therapeutic target in the allergic disorders, infectious diseases and cancers[9,10,11]. Lentivirus-mediated shRNA using lentiviral vector yielded an efficient and stable response of the target gene silencing both in vitro and in vivo[15]. It has been illustrated that sustained Ca2+ influx through Ca2+ release-activated Ca2+ channel (CRAC) requires sufficient electrical driving force which is conserved by efflux of potassium ion (K+) and intermediate Ca2+-activated K+ channel (KCa3.1, named as KCNN4) has been proven to mainly modulate the efflux of K+ and plays a pivotal role in the potassium-calcium exchange involved in the allergic and inflammatory responses[19]. The present study was aimed to explore the expression and role of KCa3.1 in the pathogenesis of AR and to further assess the therapeutic effect of lentivirus-mediated shRNA silencing of KCa3.1 in a murine AR model. P815 cell line was used to explore the role and mechanism of lentiviral KCa3.1-shRNA on mast cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
