The Roles Of PI3K/Akt/HIF-1α Pathway And Local Allergic Inflammation In The Pathogenesis Of CRSwNP

Part Ⅰ The roles of PI3K/Akt/HIF-la pathway and IL-17A in the pathogenesis of CRSwNPObjective:According to the nasal polyps,chronic rhinosinusitis(CRS)is divided into two subtypes:with polyps(CRSwNP)and without polyps(CRSsNP).The pathogenesis of CRSwNP remains unclear.According to the inflammatory cell infiltration,CRSwNP can be divided into two subgroups:eosinophilic and noneosinophilic.The Chinese subtype of CRSwNP may have a unique pathogenesis.It usually has a close relation with hypoxia,neutrophilic inflammation and Th17 cells.This study was designed to seek the role of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/hypoxia-inducible factor-1α(HIF-1α)pathway and Interleukin-17A(IL-17A)in this disease.Methods:(1)This study included a total of 53 patients with CRSwNP and 17 patients with cyst of nasal sinuses as controls,who received the functional endoscopic sinus surgery in our hospital.(2)Phadiatop test in the serum was performed.Total IgE and eosinophil cationic protein(ECP)levels in the serum were determined using the UniCAP100 automated system.(3)Total IgE level and ECP level in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA).(4)The activity of myeloperoxidase(MPO)in nasal tissues was detected by the biochemical techniques.(5)The protein expression of HIF-la,phosphorylated-Akt(p-Akt)and PI3K in nasal tissues were detected by the Western Bloting(WB)method.(6)HIF-la mRNA and IL-17A mRNA levels in nasal tissues were measured by real-time quantitative reverse transcription polymerase chain reaction(RT2-PCR).(7)We evaluated the CT scores of the CRSwNP patients using the Lund-MacKay standard.(8)We separated and cultured the fibroblasts from nasal polyps in 5 patients with CRSwNP.We used the TNF-a to stimulate and Wortmannin to inhibit the fibroblasts.Then,we examined IL-17A level by ELISA,PI3K,p-Akt and HIF-la protein levels by WB method and HIF-la mRNA and IL-17A mRNA levels by RT2-PCR.Results:(1)According to the Phadiatop test,6 patients with CRSwNP showed positive results,and controls showed all negative results.(2)The CRSwNP group showed no significant elevation in total IgE and ECP levels in the serum and supernatant,compared with the controls.(3)The CRSwNP group showed significantly elevated MPO activity in the nasal polyps,compared with the controls.(4)No significant difference was detected in the HIF-1α protein levels in the nasal tissues between the CRSwNP and control groups.(5)The CRSwNP group showed significantly elevated PI3K,p-AKT protein,HIF-1α mRNA and IL-17A mRNA levels in nasal polyps,compared with the controls.(6)In CRSwNP group,there is a positive relation among PI3K,p-Akt and HIF-1α levels.There is a positive relation between the serum and supernatant total IgE.(7)The CT scores of the CRSwNP patients had a relation with serum total IgE levels and no relation with the other indicators.(8)Stimulating by the TNF-α,the PI3K,p-AKT,HIF-1α and IL-17A levels significantly elevated in the fibroblasts.Inhibiting by the Wortmannin,the PI3K,p-AKT,HIF-1α and IL-17A levels significantly declined in the fibroblasts.Conclusion:In this study,CRSwNP mainly manifested as a neutrophilic inflammation.Systemic allergy may play a role in Chinese subjects with CRSwNP.The PI3K/Akt/HIF-la pathway played a role in the pathogenesis of the subjects with CRSwNP.IL-17A played a role in the pathogenesis of the CRSwNP patients,and controlled by the PI3K/Akt/HIF-1α pathway.The elevated IL-17A level might be responsible for the neutrophilic inflammation in the subtype of CRSwNP.PI3K/Akt/HIF-lapathway and Th17/L-17A might be new therapeutic targets for the CRSwNP.Part Ⅱ The roles of local allergic inflammation and Staphylococcus aureus enterotoxin ’superantigen’ in the pathogenesis of CRSwNPObjective:Chronic rhinosinusitis with nasal polyps(CRSwNP)is a multifactorial disease,as a result of the persistent inflammations of nasal cavity and sinuses.The pathogenesis of CRSwNP remains unclear.Local allergic inflammation plays a role in the chronic rhinitis.However,the role of local allergic inflammation in CRSwNP is still unclear.Staphylococcus aureus enterotoxins(SEs)act as superantigens in CRSwNP.But this conclusion is controversial and different in the Western country and Asian.Both local allergic inflammation and SEs can elevate the level of local sIgE in the nasal membrane.This study was designed to seek the roles of local allergic inflammation and SEs in the CRSwNP.Methods:(1)This study included a total of 18 patients with CRSwNP and 6 patients with cyst of nasal sinuses as controls,who received the functional endoscopic sinus surgery in our hospital from October to November in 2014.(2)Phadiatop test in the serum was performed.Total IgE and eosinophil cationic protein(ECP)levels in the serum were determined using the UniCAP 100 automated system.(3)Phadiatop test in the supernatant was performed.Total IgE,ECP,SEA-sIgE and SEB-sIgE levels in the supernatant were detected by the UniCAP 100 automated system.SEA,SEB and myeloperoxidase(MPO)levels in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA).(4)According to the eosinophil ratio in the pathologic sections,we divide the CRSwNP into eosinophilic subtype and noneosinophilic subtype.(5)We evaluated the CT scores of the CRSwNP patients using the Lund-MacKay standard.Results:(1)According to the results of Phadiatop test,3 patients with CRSwNP showed systemic allergies.The CRSwNP group showed no significant elevation in total IgE and ECP levels in the serum,compared with the controls.(2)2 patients with CRSwNP showed positive results of Phadiatop test in the supernatant,except the 3 patients with systemic allergies.(3)The CRSwNP group showed significantly elevated SEA and SEB levels in the supernatant,compared with the controls.But the results of SEA and SEB-sIgE tests were both negative in the supernatant of the CRSwNP patients.No significant difference was detected in the total IgE level in the supernatant between the CRSwNP and control groups.The CRSwNP group showed significantly elevated ECP and MPO levels in the supernatant,compared with the controls.(4)13 CRSwNP patients belonged to noneosinophilic subtype,others were eosinophilic subtype.(5)There was a positive correlation between the serum and the supernatant ECP levels in the CRSwNP patients.There was also a positive correlation between the serum and the supernatant total IgE levels in the CRSwNP patients.(6)The CT scores of the CRSwNP patients had no relation with the other indicators.Conclusions:(1)Systemic allergy may play a role in the CRSwNP.(2)In this study,CRSwNP mainly manifested as a neutrophilic inflammation and belonged to the noneosinophilic subtype.(3)Local allergic inflammation may play a role in the CRSwNP.(4)No evidence was shown for the role of SEs superantigens in the pathogenesis of the subjects with CRSwNP in this study.We inferred that SA might act as the other roles in the CRSwNP,but not as superantigens.