Lentiviral shRNA against KCa3.1 inhibits allergic response in allergic rhinitis and suppresses mast cell activity via PI3K/AKT signaling pathway.
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.
1] Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's hospital, Shanghai, China [2] Department of Otorhinolaryngology, Eye and ENT Hospital of Fudan University, Shanghai, China.
Hai
H
Lin
Department of Otorhinolaryngology, Eye and ENT Hospital of Fudan University, Shanghai, China.
Chunquan
C
Zheng
1] Department of Otorhinolaryngology, Eye and ENT Hospital of Fudan University, Shanghai, China [2] Department of Otorhinolaryngology, Hangzhou First People Hospital, Hanzhou, Zhejiang, China.
Jing
J
Li
1] Department of Otorhinolaryngology, Eye and ENT Hospital of Fudan University, Shanghai, China [2] Department of Otorhinolaryngology, Rui-Jin Hospital of Shanghai Jiaotong University, Shanghai, China.
Chen
C
Yang
Central Laboratory, Eye and ENT Hospital of Fudan University, Shanghai, China.
Li
L
Hu
D000818:Animals; D005260:Female; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D051058:Oncogene Protein v-akt / Q000276:immunology; D019869:Phosphatidylinositol 3-Kinases / Q000276:immunology; D034741:RNA, Small Interfering / Q000235:genetics* / Q000627:therapeutic use; D065631:Rhinitis, Allergic / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D016896:Treatment Outcome
D034741:RNA, Small Interfering; D019869:Phosphatidylinositol 3-Kinases; D051058:Oncogene Protein v-akt