Effects Of Budesonide On The Expressions Of The Glucocorticoid Receptor-alpha, Mucin-1, And Natural Immune Factor Toll-like Receptors In Nasal Polyp Epithelial Cells

Background:This study explores effects of glucocorticoid(GC)-budesonide on the expressions of glucocorticoid receptor-?,mucin-1,and natural immune factor toll-like receptors(TLR2,TLR4,TLR5,TLR7)in chronic rhinosinusitis with nasal polyps(CRSw NP),to further study the glucocorticoid(GC)-budesonide resistance mechanism of chronic sinusitis with nasal polyps.Methods:(1)Ten fresh nasal polyps specimens obtained during endoscopic sinus surgery were digested with 0.1% collagenase ? for primary culture and purification.An invert phase microscope was used for initial morphology identification of nasal epithelial cells.And successive identification with mouse anti-human cytokeratin-19 were performed by immunohistochemical study.(2)It was divided into five groups according to the budesonide action time:0 hour,6 hours,12 hours,24 hours,and 48 hours.The expressions of GR-?protein and mucin-1 protein in chronic rhinosinusitis with nasal polyps were examined by western blot,and expressions of toll-like receptors(TLR-2,TLR-4,TLR-5,and TLR-7)were examined by reverse transcription polymerase chain reaction,after training nasal polyp epithelial cells in budesonide solution.Results:(1)Western blot :0 hour,6 hours,12 hours,24 hours,and 48 hours GR-? Protein expression levels respectively is(0.818±0.693),(1.312±0.117),(0.783±0.050),(0.725±0.122),and(0.709±0.142);MUC1 Protein expression levels(1.455±0.165),(1.839±0.069),(1.021±0.182),(0.883±0.206),and(0.701±0.076).Budesonide(1×10-8 mol/L)caused up-regulation of GR-? Protein expression levels at 6h(P<0.05),compared with the 0h group;compared with the 6h group,expression of GR-?Protein was lower at 12h(P<0.05);expression of GR-?Protein at either 24 or 48 hours was not significantly different from that at 12 hours(P?0.05).Budesonide(1×10-8 mol/L)caused up-regulation of MUC1 Protein expression levels at 6h(P<0.05),compared with the 0h group;compared with the 6h group,expression of MUC1 Protein was lower at 12h(P<0.05);expression of MUC1 Protein at either 24 or 48 hours was not significantly different from that at 12 hours(P?0.05).(2)RT-PCR: 0 hour,6 hours,12 hours,24 hours,and 48 hours TLR2 mRNA expression levels respectively is(3.840±0.341),(3.015±0.358),(2.754±0.282),(5.075±0.392),and(6.083±0.363);TLR4 mRNA expression levels respectively is(0.544±0.025),(0.651±0.031),(0.407±0.024),(0.777±0.036),and(0.595±0.033);TLR5 mRNA expression levels respectively is(2.191±0.346),(1.690±0.169),(2.551±0.282),(3.049±0.278),and(1.530±0.226);TLR7 mRNA expression levels respectively is(1.680±0.119),(1.556±0.105),(1.408±0.107),(1.907±0.145),and(1.094±0.1010).Budesonide(1×10-8 mol/L)caused up-regulation of TLR2 mRNA expression levels at 6h(P<0.05),compared with the 0h group(P<0.05);there was no difference in the expression of TLR2 mRNA between 6h and 12h(P?0.05);compared with the 12 h group,there was higher expression of TLR2 mRNA at both 24 h and 48h(P<0.05).Budesonide(1×10-8 mol/L)caused up-regulation of TLR4 mRNA expression levels at 6h(P<0.05),compared with the 0h group(P<0.05);compared with the 12 h group,there was higher expression of TLR4 mRNA at 6h(P<0.05);compared with the 12 h group,there was higher expression of TLR4 mRNA at 24h(P<0.05);but there was lower expression of TLR4 mRNA at 48 h,compared with the 24 h group(P<0.05).Budesonide(1×10-8 mol/L)caused down-regulation of TLR5 mRNA expression levels at 6h(P<0.05),compared with the 0h group(P<0.05);compared with the 6h group,there was lower expression of TLR5 mRNA at both 12 h and 24h(P<0.05);compared with the 24 h group,there was lower expression of TLR5 mRNA at 48h(P<0.05).Budesonide(1×10-8 mol/L)caused down-regulation of TLR7 mRNA expression levels at 24h(P<0.05),compared with the 12 h group(P<0.05);there was no difference in the expression of TLR7 mRNA between 0h and 6h(P?0.05);there also was no difference in the expression of TLR7 mRNA between 6h and 12h(P?0.05).Conclusion:(1)The concentration of 1×10-8 mol/L of budesonide at 6 h can increase the expsession of GR-a and MUC1 protein of nasal epithelial cells,GR-a and MUC1 protein expression gradually decline as the time going,reached the lowest at 12 h.(2)The concentration of 1×10-8 mol/Lof budesonide at 6 h can down the expsession of toll-like receptors(TLR2 and TLR5)mRNA of nasal epithelial cells.But as time prolonged,toll-like receptors(TLR2 and TLR5)mRNA expression gradually rise;Here are two turning point for the expression of TLR4: at 6 h and 24 h TLR4 mRNA expression respectively raised;Within 24 h TLR7 mRNA expression began raising.(3)Under the effect of glucocorticoid(GC)-budesonide(1×10-8 M),the expression differences of GR-?,mucin-1,TLR-2,TLR-4,TLR-5,and TLR-7 in chronic rhinosinusitis with nasal polyps,suggests GR-?,mucin-1,and natural immunity in glucocorticoid resistance mechanisms of chronic sinusitis with nasal polyps playing an important role.