| Objective:To establish persistent hypoxia-induced chronic nasosinusitis mice model. Persistent hypoxia-induced oxidative stress was researched to explore whether it is an independent factor which leads to chronic nasal-sinusitis and by which causes the sequential changes of chronic nasal-sinusitis of inflammation.Methods:8-week-old C57BL6mouses were chosen for the study. Mices were exposed in the the hypoxic compartment with normal air pressure,in which oxygen concentration were controlled between8%to11%.0,2,4,6weeks after the mices were kept in the compartment,the nasal mucosa and bone morphological changes were taken and observed. Nasal mucociliary lodging and bacterial biofilm formation were observed under electron microscopy. Expression of HIF-1α, IL-17, VEGF, NF-KB were detected0,2,4,6weeks respectively after exposure.Results:1. Thanosome comparison under stereo microscope: Compared with the0-week control group, mild nasal and sinus mucosal edema were observed in hypoxia2weeks group.Evident nasal and sinus mucosal edema were found in Hypoxia four weeks group, along with blocked sinus mucosal edema. Obvious nasal and sinus mucosa along with nasal cavity congestion and swelling were observed in Hypoxia6weeks group.2.HE staining:Compared with the0-week control group, nasal and sinus inflammatory cell infiltration were observed in hypoxia2weeks group, a large number of inflammatory cell infiltration were found on nasal and sinus mucosal surface in the hypoxia4weeks group, a large number of inflammatory cell infiltration were found on the nasal and sinus mucosa the surface, inflammatory cell aggregated visibly with a large number of purulent secretions in the nasal cavity in the hypoxia six weeks group.3. Immunohistochemistry:Compared with the0-week control group, HIF-l alpha, IL-17, VEGF, NF-KB staining deepened over time with widened area in immunohistochemical sections.4. Electron microscopy:compared with the0-week control group, no obvious nasal mucociliary lodging was found in hypoxia2weeks group, nor mucosal bacterial biofilm structure. Nasal mucociliary lodging werefound in Hypoxia4weeks group with visible bacterial biofilms structure on mucosa. Obvious nasal mucociliary lodging along with some shedding mucosa and visible bacterial biofilms structure were observed in the hypoxia six weeks group.5. Western-Blot analysis:Compared with the control group at null weeks, HIF-la, IL-17, VEGF and NF-KB expression level in hypoxia nasal mucosa are increased at two weeks, there is statistically significant difference (P<0.05). they are higher at four weeks than two weeks, there were significant differences (p<0.05). Compared with4weeks, HIF-1, IL-17, VEGF, NF-KB expression level was down-regulated after six weeks, there were significant differences (p<0.05), but they are higher than the control group, with significant difference (p<0.01)6. RT-PCR analysis:Compared with the control group at null weeks, HIF-la, IL-17, VEGF and NF-KB expression level in hypoxia nasal mucosa are increased at two weeks, there is statistically significant difference (P<0.05)。they are higher at four weeks than two weeks, there were significant differences (p<0.05). Compared with4weeks, HIF-1, IL-17, VEGF, NF-KB expression level was down-regulated after six weeks, there were significant differences (p<0.05), but they are higher than the control group, with significant difference (p<0.01). The dynamic change trend of the factors expression levels are consistent with Western-Blot.Conclusion:The chronic nasal sinusitis-inflammation induced by continuous hypoxia has time sequence. the inflammation is presented earliest at2weeks, comes to the top at4weeks and is down-regulated at6weeks, but still significantly higher than the control group at null week. This shows that nasal mucosa for hypoxia begins with the tolerance after4weeks, but the inflammation is irreversible. Objective:To detect the dynamic expression levels of the factors related to the Mucosal inflammation at differently anatomical location in different anatomical location andexplore the differences of the dynamic factor expression related to mucosal inflammationt in the process of the chronic rhinosinusitis of the mouse induced by chronic hypoxia.Methods:The8weeks old C57BL6mouse are selected as the research object, feed in the normobaric hypoxia cabin, which of oxygen concentration are controlled from8%to11%. The samples on the mucosa and the bone specimens are collected from4anatomical sites (nasal septum, nasal mucosa, middle turbinate mucosa of maxillary sinus mucosa, mucosa).by placing the point time of time point detection of expression changes on the HIF-1a, IL-17, VEGF and NF-KB factors are detected respectively after0,2,4,6weeks and the double fluorescent staining is carryed on.Results:1、RT-PCR analysis:1), The same time point nasal mucosa of the anatomical location of HIF-1alpha, IL-17, VEGF, NF-KB expression levels SB<BN<ZN<SN, P values less than0.05, statistically significant. And the difference was no change in trend0,2,4,6weeks4time points.2). Mucous membranes of the nasal anatomical location of HIF-1alpha, IL-17,VEGF, NF-KB expression level of With continued hypoxia stimulate the performance of the same expression trend, that is0-4weeks expression levels continued to rise after4weeks lowered, P<0.05, but the expression level of the factor6weeks hypoxic still significantly higher than the0-week control group.2, Western-Blot analysis:1), The same time point nasal mucosa of different anatomical location of HIF-1alpha, IL-17, VEGF, NF-KB expression levels SB<BN<ZN<SN, P values less than0.05, a significant difference. And the difference was no change in trend0,2,4,6weeks4time points.2), The nasal anatomical location mucosa of HIF-1alpha, IL17, VEGF, NF-KB expression levels with sustained hypoxia stimulate showed the same trend of expression, i.e.0-4weeks expression levels continued to rise, weeks after the cut, P<0.05, but the6weeks of hypoxia still significantly higher than the expression level of the factor0-week control group.3, immunofluorescence double staining:HIF-1alpha and IL-17, VEGF, NF-KB coexist on the nasal mucosa membrane and the periosteal surface, with sustained hypoxic stimulation time variation the the double subscript coexist factor dyeing darker, a broad range of variableConclusion:1The dynamic change trend of expression levels on the HIF-la, IL-17, VEGF and NF-KB factors of the nasal mucosa at the differently anatomical locations are consistent. In other words, the time sequence change is identical.the factors expression levels are increased from null to4weeks, are down-regulated after4weeks and are still higher than that of the control group at6weeks2, The HIF-1a, IL-17, VEGF and NF-KB factors expression levels of the differently anatomical location are differences. This difference does not change at4timespoints. It is not affected by the time changes namely and only related to the different anatomical locations... |
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