Preliminary study on Aspergillus fumigatus-induced IKKα controlled maspin protein expression in respiratory epithelial cells of rats / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study if the expression of maspin protein in respiratory epithelial cells was downregulated through IκB kinase-α (IKKα)-controlled mechanism in an Aspergillus fumigatus-induced model in rat.</p><p><b>METHODS</b>Inactivated Aspergillus fumigatus hyphae (AFH) suspension was used to induce respiratory epithelial cells (REC) cultured in vitro in rat, with PBS buffer as control. By RT-PCR, the mRNA expression of maspin was quantified, and by immunocytochemistry, the expression of maspin and IKKα in REC was observed. Furthermore AFH (from level 1 to level 3) suspension was prepared to induce REC. Then Western blot hybridization technique was used to detect the expression of maspin and IKKα protein. All data were processed by analysis of variance and two-variable correlation analysis.</p><p><b>RESULTS</b>By RT-PCR, a statistically significant (t = 2.463, P < 0.05) expression of maspin mRNA was found (0.128 ± 0.059 in AFH group, 2.972 ± 0.353 in control group). By Immunocytochemistry, the difference of maspin protein color in different groups was shown statistically significant in integrated scoring (t = 3.721, P < 0.05, weak positive in AFH group, moderately positive in control group). While in IKKα color study, the difference between the two groups was also statistically significant (t = 6.825, P < 0.05) in integrated scoring, with a moderate positive in AFH group and weak positive in control group. By Western Blot hybridization, grayscale ratio of maspin and β-actin was 0.912 ± 0.023 in control group, 0.607 ± 0.030, 0.476 ± 0.019, 0.416 ± 0.017 in AFH 1-3 groups, respectively, with a statistically significant difference within the four groups (F = 281.91, P < 0.05); While grayscale ratio of IKKα and β-actin was 0.624 ± 0.012 in control group, 0.739 ± 0.020, 0.778 ± 0.010, 0.927 ± 0.017, respectively, in AFH 1-3 groups; with a statistically significant difference within the four groups(F = 200.91, P < 0.05). Moreover, the difference between any two groups from both AFH group (including subgroup 1, 2 and 3) and control group was statistically significant. Two-variable correlation analysis indicated a negative correlation between expression of maspin and IKKα (r = -0.911, P < 0.05).</p><p><b>CONCLUSIONS</b>Induced by Aspergillus fumigatus, the rat respiratory epithelia might upregulate the expression of IKKα with a downregulated expression of maspin protein.</p>

Effects of sulphur dioxide inhalation on allergic rhinitis in mice / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the effect of sulphur dioxide (SO2) inhalation on the pathogenesis of allergic rhinitis (AR) in mice, and investigate the toxic effect of SO2 on respiratory tract mucosa.</p><p><b>METHODS</b>Fifty male Kunming strain mice were randomly allocated to five groups, i. e., control group, AR group and AR plus different concentrations of SO2 group (three sub-groups). Nasal cavity irrigating solution was gathered from nasal cavity, and blood from the orbital venous sinus after anesthesia The concentrations of IL-5 and IL-13 in the peripheral serum and nasal cavity irrigating solution were measured by ELISA. The number of eosinophils (EOS) per square millimeter in sinus mucosa was calculated by hematoxylin-eosin. The expression of SP-D in nasal mucosa was analyzed by immunohistochemistry technique.</p><p><b>RESULTS</b>With increasing concentrations of SO2, the levels of IL-5 and IL-13 in the peripheral serum, and the density of Eos in sinus mucosa increased simultaneously. A positive correlation existed between the concentration of inhaled SO2 and the elevation of both IL-5, IL-13 and Eos infiltration in nasal mucosa. The coefficient correlation relatively were 0.894, 0.874, 0.894, 0.891 and 0.870 (P <0.01). The expression of SP-D in 56 mg/m3 and 112 mg/m3 SO2 groups was higher, while it was lower in 168 mg/m3 SO2 group (P < 0. 001).</p><p><b>CONCLUSIONS</b>The study showed that sulphur dioxide inhalation facilitates the onset of allergic rhinitis in mice. SO2-related Th2-derived cytokines as well as the infiltration of EOS in nasal mucosa help to aggravate the development of allergic rhinitis.</p>

Preliminary investigation into the allergic rhinitis complicated with acute bacterial sinusitis in mice / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To develop a mouse model of bacterial rhinosinusitis superposed on allergic rhinitis (AR), and to explore whether ongoing allergic rhinitis enhance the acute sinus infection and inflammation associated with Streptococcus pneumoniae (SP).</p><p><b>METHODS</b>Fourty mice of C57BL6/J were randomly divided on average into 4 groups: A [ovalbumin (OVA) + SP], B [OVA + normal saline (NS)], C [phosphate buffered solution (PBS) + SP] and D (PBS + NS). (1) Group A and B were sensitized by intraperitoneal injection with 200 microl (10%) OVA on days 1 through 9, and exposed to OVA (6%) intranasally on days 10 through 17, to induce allergic inflammation. OVA was replaced with PBS in group C and D in the same way. (2) Subsequently, group A and C were inoculated with SP intranasally on day 13, and NS was used in group B and D. On the 6th day after inoculation, mice were killed. Blood was collected from the orbital venous sinus after anesthesia. The heads were embedded with paraffin and serial sections were followed and stained with hematoxylin-eosin and toluidine blue (0.5%) for histological analysis and inflammation cells count. The number of polymorphonuclear neutrophils (PMN) and eosinophils (EOS) per square millimeter of sinus mucosa were calculated by using a computer-aided special software under microscope.</p><p><b>RESULTS</b>AR models were successfully established in 9 mice from group A and 8 from group B. Histologic examination of the sinus from group A and B revealed significant mucosal edema and dilated venules. The symptoms were mild in group C, and no symptom was observed in group D. PMN (x +/- s) in group A (139.3 +/- 26.5)/mm2 was significantly higher than that in group B (70.7 +/- 16.7)/mm2, C (63.0 +/- 14.7)/mm2 and D (40.2 +/- 14.1)/mm2 respectively (P < 0.01); EOS and serous IL-5 level in group A (134.6 +/- 25.5)/mm2, (48.2 +/- 13.9) pg/ml and B (116.2 +/- 25.2)/mm2, (40.8 +/- 7.8) pg/ml, were higher than that in group C (16.7 +/- 2.7)/mm2, (23.9 +/- 8.7) pg/ml (P < 0.05) and D (13.4 +/- 4.9)/mm2, (24.6 +/- 6.5) pg/ml (P < 0.05).</p><p><b>CONCLUSIONS</b>The data demonstrate that an ongoing local allergic response augments bacterial infection in mice, and allergic sensitization alone without SP does not induce the sinus infection.</p>

Alteration of cytokines and mucin in lower respiratory tract in allergic rhinitis model in rats / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the relationship between allergic rhinitis (AR) and asthma as well as the mechanisms related with it.</p><p><b>METHODS</b>Sixty healthy rats were randomly divided into AR group and control group. AR model was established by intraperitoneal injection of ovalbumin (OVA) and nasal challenge with OVA. Nasal mucosa and lung tissue from both groups were stained with hematoxylin-eosin (HE), alcian-blue and periodic acid-schiff (AB-PAS), respectively. At the same time, the lung tissue was studied by transmission electron microscopy (TEM). Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to examine the level of interleukin-4 (IL-4) in bronchial alveolus lavage fluid (BALF) and the expression of intercellular adhesion molecule-1 (ICAM-1) and MUC5AC in nasal and lung tissue, respectively.</p><p><b>RESULTS</b>Infiltration of inflammatory cells in nasal mucosa and lung tissue of AR model in rat was evident. Cilia destruction of bronchial epithelial cells of AR model was found. The level of IL4 in BALF of AR group (58.10 +/- 7.92) pg/ml was significant higher compared with that in control group (24.66 +/- 2.07) pg/ml. The expression of ICAM-1 (0.66 +/- 0.24) and MUC5AC (0.71 +/- 0.10) in lung tissue were both significantly higher than that in control group (0.23 +/- 0.02, 0.29 +/- 0.03).</p><p><b>CONCLUSIONS</b>Allergic inflammation in nasal mucosa not only leads to changes in both histopathology and immunology, but also initiates the inflammation in lower respiratory tract mainly causing the change of cytokines and mucin.</p>