Mechanism of action of formononetin in alleviating allergic asthma through DRP1-NLRP3 signaling pathway / 中国药理学通报

Aim To investigate the mechanism by which formononetin (FN) inhibits mitochondrial dynamic-related protein 1 (DRP1) -NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation. Methods In order to establish allergic asthma mouse model, 50 BALB/c mice aged 8 weeks were divided into the control group, model group, FN treatment group and dexamethasone group after ovalbumin (OVA) induction. Airway inflammation and collagen deposition were detected by HampE and Masson staining. Th2 cytokines and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and IgE levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA, ROS in BEAS-2B cells was assessed by DCFH-DA staining, DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence, and the DRP1-NLRP3 pathway was analyzed by immunoblotting. Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model, including reducing eosinophil accumulation, airway collagen deposition, decreasing Th2 cytokine and IgE levels, reducing ROS and MDA production, increasing SOD and CAT activities, and regulating DRP1-NLRP3 pathway-related protein expression, thereby relieving inflammation. Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.

Panax notoginseng saponin Rl attenuates allergic rhinitis through AMPK/DRP1 mediated mitochondrial fission / 中国药理学通报

Aim To investigate whether notoginsenoside Rl (PNS-R1) alleviates allergic rhinitis (AR) through AMP-activated protein kinase (AMPK)/mitochondrial fission critical protein (DRP1) -mediated mitochondrial fission. Methods Different doses of PNSRl were used to treat ovalbumin (OVA) -induced AR model mice,and the inhibitory effect of PNS-R1 on AR was investigated by observing allergic symptoms such as nasal rubbing and sneezing, as well as HE staining of nasal tissues. Serum IgE levels and nasal lavage fluid (NLF) inflammatory cytokine levels were detected by enzyme-linked immunosorbent assay (ELISA) and apoptosis-related proteins were detected by Western blot. In vitro human nasal epithelial cells (HNEpC) were stimulated with IL-13 to observe apoptosis, mitochondrial membrane potential, cellular ROS and mitochondrial ROS production, as well as the expression levels of AMPK/DRP1, expression levels of the TXNIP/NLRP3 inflammasomes and the translocation of DRP1. Results PNS-R1 attenuated allergic symptoms in AR mice, HE staining reduced inflammatory cells and reduced the levels of OVA-specific IgE in serum, and the levels of IL-4, IL-6, and IL-8 in NLF. PNS-R1 attenuated the apoptosis and ROS production of nasal epithelial cells in AR. In vitro PNS-R1 could up-regulate mitochondrial membrane potential after IL-13 stimulation, reduce ROS and mtROS production, the proportion of apoptotic positive cells, and reduce cleaved caspase-3, Bax, and up-regulate Bcl-2 expression, down-regulate DRP1 phosphorylation (Ser 616) and DRP1 translocation at the mitochondrial membrane in an AMPK-dependent manner, reducing TXNIP/NLRP3 expression. Conclusions PNS-R1 can protect mitochondrial integrity by inhibiting the AMPK/DRP1 signaling axis and its subsequent TXNIP/NLRP3 signaling axis,thereby alleviating rhinitis in AR mice.

JTE-013 mediates RhoA/ROCKl/Drpl signaling axis regulating mitochondrial damage and apoptosis to alleviate allergic rhinitis / 中国药理学通报

Aim To investigate the protective effect and mechanism of JTE-013 on allergic rhinitis (AR) by regulating mitochondrial injury and apoptosis through RhoA/ROCKl/Drpl pathway. Methods AR model was established by ovalbumin (OVA) in mice. Nasal tissue sections were then stained with HE, TUNEL and DHE. Western blot assay. In vitro, human nasal epithelial cells (HNEpCs) were stimulated with human recombinant interleukin-13 (IL-13), and the effects of JTE-013 and Y27632-related protein expression were detected by Western blot. Immunofluorescence was used to observe the effects of JTE-013 and Y 27632 on total ROS, mitochondrial membrane potential and mitochondrial ROS generation, Drpl translocation and Cyt-c expression in cells. Results JTE-013 reduced the frequency of nose rubbing and sneezing, reduced nasal mucosal thickening and decreased eosinophil infiltration in AR mice. TUNEL and DHE staining results suggested that JTE-013 could inhibit apoptosis and reduce ROS expression in mouse nasal epithelial cells. Western blot showed that both JTE-013 and Y 27632 could significantly reduce RhoA, ROCK1, Drpl and p-Drpl(616), inhibit the expression of apoptotic proteins Bax, cleaved-caspase-3, Cyt-c, cleavedcaspase-9 and up-regulate the expression of p-Drpl (637) and Bcl-2. Immunofluorescence showed that inhibitors of JTE-013 or ROCK1 almost blocked IL-13mediated increase in ROS and mtROS production, inhibited decrease in mitochondrial membrane potential, and blocked Cyt-c expression and Drpl translocation in nasal mucosal epithelial cells. Conclusion JTE-013 can regulate the morphology and function of mitochondria by inhibiting RhoA/ROCKl/Drpl signaling axis, thereby alleviating nasal epithelial cell inflammation in mice with allergic rhinitis.

Polydatin alleviates allergic rhinitis by regulating PINK1-Parkin-mediated mitophagy / 中国药理学通报

Objective To investigate the protective effect and mechanism of polydatin ( PD) on allergic rhinitis (AH) by regulating mitophagy through PINK1- Parkin signaling pathway, and to provide a new target for clinical treatment of AH.Methods Thirty-two BALB/c murine were randomly divided into 4 groups: control group, OVA group, PD low-dose (30 mg • kg 1 ) and high-dose ( 45 mg • kg 1 ) treatment groups.At the end of modeling, the total number of sneezing and nasal nibbing of murine was recorded.HE staining was used to observe the morphology of na- sal mucosal epithelium and eosinophil infiltration.Western blot was used to detect the expression of P1NK1 , Parkin, TOM20 and mitochondrial apoptosis- related proteins Bax, Bcl-2, caspase-3, cleaved- caspase-3 and Cytochrome C.Hie expression of P1NK1 and cleaved-caspase-3 in nasal epithelial cells (HNEpC) was observed by immunofluorescence.Re¬sults The frequency of sneezing and nasal rubbing movements was significantly increased, the nasal mu¬cosa epithelium was thickened, and eosinophils were accumulated in AH murine , these results were reversed after PD treatment.Western blot results shower] that signaling proteins PINK1/Parkin anrl pro-apoptotie pro¬teins, including Bax, caspase-3, cleaved-caspase-3 and Cytochrome C were significantly overexpressed, the expression of TOM20 and Bcl-2 was decreased in OVA group, and PD up-regulated the levels of P1NK1 , Parkin, as well as Bcl-2 and inhibited the expression of TDM20 and pro-apoptotic proteins, while after pre- treatment with mitochondrial division inhibitor 1 ( Mdi- vi-1 ) , the expression of P1NK1 and Parkin was re¬duced , the expression of TOM20 was increased, while PD treatment did not significantly affect this effect.From the immunofluorescence results, it can be seen that the level of P1NK1 was increased after IL-13 stim¬ulation of HNEpCs compared with the control group, and PD further up-regulated the expression of P1NK1 , which was suppressed after pretreatment with Mdivi-1 , while PD did not change this phenomenon.Western blot results for P1NK1 and cleaved-caspase-3 were con¬firmed by immunofluorescence.Conclusion PD may activate mitophagy through the P1NK1 -Parkin signaling pathway, thereby protecting against AR.

Nasality Changes With Age in Normal Korean-Speaking Adults

OBJECTIVES: This study was performed to investigate the effects of aging on nasality and the influence of age-related changes in nasal cavity volume and nasal patency on nasality. METHODS: A total of 180 healthy Korean-speaking adult volunteers, who had no nasal or voice-related complaints, were enrolled in this study. Nasometry, acoustic rhinometry, and rhinomanometry were performed to obtain the nasalance score, nasal cavity volume, and nasal resistance, respectively. Changes in these parameters with age were analyzed. RESULTS: Nasal cavity volume increased significantly, and nasal resistance decreased significantly, with age. The nasalance scores for the nasal passage and oronasal passage decreased significantly with age, while there were no age-related changes in nasalance scores for the oral passage. CONCLUSION: Nasalance scores for the passages containing nasal consonants decreased with age although significant increases were observed in nasal cavity volume and nasal patency with age. Therefore, the age-related decreases in nasalance scores may result from factors other than changes in the nasal cavity.

Effects of Repeated Ozone Exposure on Allergic Responses in Mice / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: We Investigated whether repeated ozone exposure has a priming effect on allergic sensitization and whether ozone exposure has an exacerbating effect on allergic responses during allergen challenge in previously sensitized animal. SUBJECTS AND METHOD: Thirty-five female BALB/c mice were divided into the following five groups; controls (I), ovalbumin (OVA)-aerosol exposure group (II), OVA-aerosol/ozone exposure group (III), OVA-systemic sensitization group (IV), and OVA-systemic sensitization/ozone exposure group (V). Mice of group III and V were exposed to 0.3 ppm ozone for 4 hours, three times a week for four weeks. Mice of group II and III were exposed to OVA-aerosol for 20 minutes, five times a week for four weeks. Mice of group IV and V were sensitized by intraperitoneal injections with OVA/Al(OH)3 solution on day 0, 7, and 14. Animals received five consecutive allergen challenges by OVA-aerosol nebulization from day 23 to 27. At 24 hours after the last exposure, nasal lavage fluid (NLF) and blood were obtained. The concentrations of total serum IgE, anti-OVA IgE, and anti-OVA IgG1 were measured by enzyme-linked immunosorbent assay (ELISA). The levels of IL-4, IL-5 and INF-gamma in NLF were measured by ELISA. In addition, Luna staining was performed to identify eosinophils that infiltrated in nasal mucosa. RESULTS: OVA-aerosol exposure alone induced weak allergic response to OVA in nonsensitized mice. In contrast, the combination of OVA-aerosol and ozone resulted in augmented immune responses to OVA as indicated by significant increases in total serum IgE, anti-OVA IgE, and anti-OVA IgG1 antibody titers, as well as significant increases in the levels of Th2 cytokines In NLF and the number of eosinophils infiltrating nasal mucosa. Although the titers of total IgE and anti-OVA IgE of group III were significantly lower than those of group IV, the titers of anti-OVA IgG1, levels of IL-4 and IL-5, and the number of eosinophIls infiltrating nasal mucosa showed no significant differences between group III and group IV. In sensitized mice, ozone exposure durIng OVA challenge enhanced the allergic responses to OVA. CONCLUSION: These results suggest that ozone exposure induces and enhances the allergic responses by augmenting the production of allergen-specific antibody and Th2 cytokines.

Effects of Ebselen on Ozone-Induced Nasal Mucosal Inflammation in Mice / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Although the mechanism of ozone-induced airway inflammation and hyperresponsiveness is largely unknown, NO and peroxynitrite has been suggested to be associated with it. Ebselen, a seleno-organic compound, is known to inhibit the production of superoxide, iNOS-related NO, and their combined product, peroxynitrite. The purpose of this study is to investigate whether ebselen suppress ozone-induced nasal inflammation and whether ebselen inhibit the production of NO and peroxynitrite in nasal mucosa. SUBJECTS AND METHOD: Thirty-six BALB/c mice were divided into three groups: control group, ozone exposure group, and ozone+ebselen treated group. In the ozone exposure group, mice were exposed to 1 ppm ozone for 8 hours a day for 3 consecutive days. In the ebselen treated group, the ebselen (32.5 mg/kg) solution was injected intraperitoneally 1 hour before and 3 hours after the ozone exposure. At 18 hours of the last ozone exposure, Evans blue was infused via tail vein in 6 animals of each group. Mice were sacrificed five minutes later and nasal mucosa was obtained to measure the amount of extravasated Evans blue dye. From the remaining 6 animals in each group, nasal lavage fluid (NLF) was obtained to measure the concentration of albumin and the number of neutrophils. After lavage fluid was obtained, nasal mucosa was taken for immunohistochemical staining against iNOS and nitrotyrosine usng the ABC method. RESULTS: Extravasation of Evans blue was significantly increased in the ozone exposure group, but it was significantly decreased in the ebselen treated group. Albumin concentration in NLF showed a tendency to increase in the ozone exposure group and a tendency to decrease in the ebselen treated group when compared with the ozone exposure group. The number of neutrophils was significantly increased in the ozone exposure group and was decreased more in the ebselen treated group than in the ozone exposure group. Immunoreactivity to iNOS and nitrotyrosine was strongly expressed in nasal mucosa of the ozone exposure group. However, it was nearly abolished by the treatment with ebselen. CONCLUSIONS: These results may suggest that ebselen can be applied as a useful therapeutic agent for airway diseases by modulating the oxidant-related inflammatory process.

Repeated Ozone Exposure Induces Th2 Immune Responses in Mice

BACKGROUND AND OBJECTIVES: It has been reported that ozone exposure exacerbate allergic rhinitis symptoms and may contribute to increase allergic rhinitis prevalence. However, a causal relationship still remained unsolved. The purpose of this study is to investigate whether prolonged exposure to ozone induce Th2 immune response without allergen. MATERIALS AND METHODS: Fourteen BALB/c mice were divided into two groups: control group and ozone exposure group. Mice were exposed to 0.3 ppm of ozone for 4 hours a day, 3 times per week, for 4 weeks. At 24 hour after the last ozone exposure, nasal lavage fluid (NLF) was obtained to measure the levels of cytokine IL-4, IL-5, and IFN-gamma. After lavage fluid was obtained, blood was obtained via inferior vena cava to measure the amount of total IgE and IgG1. The concentration of cytokines and immunoglobulins was measured using the ELISA method. In addition, Luna staining was performed to identify eosinophils infiltrated in nasal mucosa. RESULTS: The levels of IL-4 and IL-5 in NLF were significantly increased in ozone exposure group compared with control group. But the level of IFN-gamma in NLF shows no significant difference between two groups. Serum total IgE and IgG1 were significantly increased in ozone exposure group compared with control group. On histologic examination, number of eosinophils infiltrating nasal mucosa was significantly increased in ozone exposure group. CONCLUSION: These results suggest that repeated ozone exposure induces Th2 response in the nasal mucosa of mice.

The Inhibitory Effect of Anti-P-Selectin, Anti-PSGL-1 and Anti-CD49d Antibodies on Experimental Allergic Airway Inflammation in Mice

BACKGROUND AND OBJECTIVES: Eosinophil recruitment in allergic inflammation is dependent on the interactions between adhesion molecules. The objective of this study was to determine whether blocking of P-selectin glycoprotein ligand-1 (PSGL-1), P-selectin or CD49d adhesion molecules using monoclonal antibodies can reduce eosinophil recruitment in mice with the airway allergy. MATERIALS AND METHODS: Seven different groups of mice were used. One experimental group of mice (group A) served as naive control and sensitized with phosphate-buffered saline (PBS). Six experimental groups of mice (groups B to G) were sensitized to ovalbumin (OVA). The mice in group B were treated with PBS before OVA aerosol challenge and served as positive controls. The mice in other remaining groups were treated with monoclonal antibodies (mAb) against specific adhesion molecules before OVA challenge; anti-P-selectin mAb (group C), anti-PSGL-1 mAb (group D), anti-CD49d mAb (group E), anti-P-selectin mAb & anti-CD49d mAb (group F), anti PSGL-1 mAb & anti-CD49d mAb (group G). Eosinophils in the nasal mucosa and bronchoalveolar lavage (BAL) fluid in each animal group were measured. RESULTS: Mucosal eosinophilic infiltrations were significantly reduced at the mice in the group C, F or G compared with group B. Eosinophils in BAL fluid were significantly reduced only at the group C mice. CONCLUSION: These results suggest that blockade of P-selectin is superior to blockade of PSGL-1 or CD49d in the inhibitory effect against eosinophil recruitment.

Anatomic Changes of Intranasal and Surrounding Bony Structures in Patients with Nasal Septal Deviation: A CT Analysis / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: In the cases of nasal septal deviation (NSD), bilateral nasal structures, such as inferior turbinate, nasal floor, or lateral nasal wall, are often asymmetric. These patterns may be somewhat uniform according to the direction of septal deviation. The aims of this study are to establish the anatomical patterns of the intranasal and surrounding bony structures in the cases of NSD by using radiologic measurement. SUBJECTS AND METHOD: We took OMU CT scans of 38 adult non-traumatic NSD patients without concurrent sinonasal inflammation. For each patient, we selected 4 consecutive coronal images of the most severely deviated portion for the measurement. We set various parameters which could reflect the anatomic patterns of the bony structures, then measured them in the selected images. RESULTS: The levels of bilateral nasal floors were not the same, and the floor of nasal cavity was significantly declined to convex side. The angle of lateral nasal wall was significantly larger in the concave side than in the convex side. The horizontal width or the angle of the inferior turbinate was significantly larger in the concave side than in the convex side. CONCLUSION: In the cases of NSD, the symmetry of bilateral intranasal or surrounding bony structures was altered significantly, and its pattern was uniform according to the direction of deviation. These findings suggest that non-traumatic NSD may be the result of asymmetric growth of facial bones.

The Effects of Adenoidectomy on Nasal Airway Resistance and Nasal Geometry

The primary objective of this study was to determine whether adenoid hypertrophy and subsequent adenoidectomy affect pediatric nasal airway resistance and nasal geometry. The secondary objective was to evaluate the relationships between the degree of adenoid hypertrophy and nasal airway resistance or nasal geometry. Fifty-one children, aged 5 to 10 years, selected for adenoidectomy due to chronic nasal obstruction and mouth breathing were enrolled. The size of adenoid was evaluated by cephalometric radiograph. Nasal airway resistance and nasal geometry were evaluated by active anterior rhinomanometry and acoustic rhinometry respectively. These measurements were repeated 3 months after operation. The size of adenoid was found well correlated to preoperative nasal airway resistance but was not to preoperative nasal geometry. Nasal geometry was not changed after operation. However, nasal airway resistance was reduced significantly at 3 months after operation and the size of adenoid was found well correlated to postoperative changes of nasal airway resistances.

The Effects of Celecoxib on Wound Healing in Murine Wound Models / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Celecoxib has suppressive effects on the growth, angiogenesis, metastasis of solid tumors including head and neck squamous cell carcinoma. Recent report suggests that celecoxib can also be usefully applied for preventing tumor recurrence in the postoperative conditions with possible residual tumors. The aim of this study is to investigate the effects of celecoxib on the post-surgical wound healing and the systems including the gastro-intestinal (GI) tracts. MATERIALS AND METHOD: Incisional and excisional wound models were created in the C3H mice and celecoxib was administered at a dose of 20 mg/kg/day to the wounded mice. Photographic documentation of the wounds was performed every week. The mice were serially sacrificed 3, 7, 14, and 28 days after wounding. The re-epithelialization and capillary number of the wounded skin were measured and the side effects of celecoxib were observed. RESULTS: Re-epithelialization was suppressed by celecoxib only in the early phase at the day 10 of wounding, which was all recovered in the late phase at day 14. The capillary number of the wounded bed was not affected by the celecoxib treatment. In addition, celecoxib had no significant side effects on the body weight change and the GI tracts of the wounded mice. CONCLUSION: This murine wound models suggest that celecoxib is a safe drug with no significant side effects to treat late wound healing or the GI tracts.