Experimental Studies Of The Regulation Of Calcium-Activatied Chloride Channel 1 On MUC5AC Expression In Chronic Rhinosinusitis

Part I Experimental Studies Of The Relationship Between Calcium-Activatied Chloride Channel 1 And Chronic RhinosinusitisObjective:To exmatine the expression of calcium-activatied chloride channe 1 messenger RNA(mRNA) and proteins in human sinus mucosa of chronic rhinosinusitis without nasal polyposis (CRS) and chronic rhinosinusitis with nasal polyposis(CRS/NP). Methods: Ethmoid sinus mucosa was harvested from patients undergoing endoscopic sinus surgery for CRS with and without nasal polyposis and non-CRS pathologies (control).Then sinus mucosa was analyzed using real-time polymerase chain reaction (real-time PCR) to detect the mRNA of calcium-activatied chloride channe 1. Immunofluorescent staining was used to evaluate the expression of calcium -activatied chloride channe 1 proteins in the sinus mucosa. Area ratio of positive cells in the epithelia was compared among the CRS,CRS/NP and the control groups. Results:the mRNA of calcium -activatied chloride channe 1 in the sinus mucosa of CRS and CRS/NP significantly increased compared with that in normal sinus mucosa (p< 0.01). and no significant difference was found between the mucosa of CRS and that of CRS/NP (p>0.05). Calcium-activatied chloride channel 1 proteins were expressed mainly in the goblet cells, hyperplasia of goblet cells were observed in all cases of CRS and CRS/NP. Area ratio of positive cells in the epithelia was found no different between the CRS group and the CRS/NP group (p> 0.05), whereas it was significantly lower in the normal group compared with the other two groups, respectively (p 0.01).Conclusions:This study showed that calcium -activatied chloride channel 1 gene was up-regulated in sinus mucosa of CRS and CRS/NP. Calcium -activatied chloride channel 1 may play an important role in the pathogenesis of CRS and CRS/NP for the over-production of MUC.Part II Experimental Investagation On The Relationship Between MUC5AC And Chronic RhinosinusitisObjective:To investagate the expression of MUC5AC messenger RNA(mRNA) and protein in human sinus mucosa of chronic rhinosinusitis with and without nasal polyposis (CRS and CRS/NP). Methods:Ethmoid sinus mucosa was obtained from patients with CRS with and without nasal polyposis undergoing endoscopic sinus surgery and non-CRS pathologies (control).Then sinus mucosa was analyzed using real-time polymerase chain reaction (real-time PCR) to examine the mRNA of MUC5AC. Immunofluorescent staining was used to evaluate the expression of MUC5AC protein in the sinus mucosa. Area tatios of positive cells in the epithelia were compared among the CRS, CRS/NP and normal groups. Results:Expressions of mRNA of MUC5AC in the sinus mucosa of CRS and CRS/NP significantly increased compared with that in normal sinus mucosa (p< 0.01), and no significant difference was found between the mucosa of CRS and that of CRS/NP (p> 0.05). MUC5AC protein expressed mainly in the goblet cells, hyperplasia of goblet cells was observed in all cases of CRS and CRS/NP. There was no difference of area tatios of positive cells in the epithelia founded between the CRS group and the CRS/NPgroup (p> 0.05), whereas it was significantly lower in the normal group compared with the other two groups, respectively (p< 0.01).Conclusions:These results suggested that MUC5AC gene up-regulatation in sinus mucosa of CRS and CRS/NP may play an important role in the pathogenesis of CRS and NP associated with the hyperplasia of goblet cells and mucin overproduction.Part III Study Of The Regulation Of Calcium-Activatied Chloride Channel L On MUC5AC Expression In Sinus Mucosa Tissue CultureObjective:To investigate the regulation of CaCClon MUC5AC expression in sinus mucosa tissue culture. Methods:mucosa (sinus) tissures obtained from patients with chronic rninosinusitis undergoing sinus surgery were cultured for 24 hours.every specimen was divided into 5 groups, A group:maintained in medium without adding any stimulators; group B:added with recombinant human (rh)TNF-a (10 ng/mL); group C:added with CaCC1 inhibitor niflumic acid lOumol/L followed by TNF-a (10 ng/mL) 30 minutes later. group D:added with CaCC1 inhibitor niflumic acid 100 umol/L followed by TNF-a (10 ng/mL) 30 minutes later:group E:added with CaCC1 inhibitor niflumic acid 500 umol/L followed by TNF-a (10ng/mL) 30 minutes later. CaCC1 and MUC5AC protein expressions were quantified by using Western blotting. Results:TNF-a significantly increased CaCCl and MUC5AC proteins expressions in the mucosal explant tissue (P<0.01). Inhibition of CaCC1 with niflumic acid showed a significant dose-dependent reduction of CaCCl and MUC5AC proteins in the mucosal explant tissue (P<.05). Conclusions:TNF-a induced MUC5AC protein expression could be decreased by niflumic acid in explant sinus mucosa tissue which certificated that CaCC1 regulated MUC5AC expression in humann sinus mucosa tissure culture.