OBJECTIVE: To examine AMCase mRNA expression levels in nasal mucosa of allergic rhinitis SD rats. METHOD: Thirty SD rats were chosen and randomly divided into the experimental group and the control group. 20 in experimental group and 10 in the control group. AR model rats were established through repeated intraperitoneal shot of ovalbumin (OVA) for 2 weeks and consequently confirmed by local challenge with OVA for 1 week. The control group was treated by the same method with Physiological saline water instead of OVA. After the last excitation allergic rhinitis was diagnosed according to the accumulation score about nasal symptom. The septal mucosa of all rats were used to diagnose pathologically by HE dyeing. AMCase mRNA in nasal mucosa, obtained from the bilateral nasal mucosa in two groups, were used to do reverse transcriptive polymerase chain reaction. Real-time polymerase chain reaction was used to examine AMCase mRNA expression levels. RESULT: (1) The results showed definitely that there were positive expression of AMCase mRNA in normal nasal mucosa. This expression increase significantly during nasal allergy (P < 0.05). (2) The increased expression of AMCase mRNA in allergic rhinitis are related with the nasal symptoms score (r = 0.411, P < 0.05). CONCLUSION Increased expression of AMCase mRNA in nasal mucosa in allergic rhinitis model might play roles in the pathogenesis of AR, and Restrain the enzyme activity could become new treatment targets of allergic rhinitis.
Animals ; Chitinases ; metabolism ; Nasal Mucosa ; metabolism ; Ovalbumin ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic ; enzymology ; metabolism

Local anesthetics at a concentration of 10 mM(procaine and lidocaine) were found to inhibit competitively Ca++ binding to lipidextracted RBC membrane, and also to egg albumin film fixed on cover glasses or impregnated into Whatman filter paper (No. 1). A competitive inhibition by local anesthetics was also found in Ca++ binding to Whatman filter paper which had been pretreated with organic solvents to extract possible contaminated lipids. Therefore, it is suggested that the local anesthetics inhibit Ca++ binding not only to phospholipids but to some macromolecules such as membrane proteins, egg albumin film and filter paper.
Anesthetics, Local/pharmacology* ; Calcium/metabolism* ; Cell Membrane Permeability/drug effects ; Erythrocytes/cytology* ; Filtration ; Human ; Lidocaine/pharmacology ; Ovalbumin/metabolism* ; Procaine/pharmacology

To explore the relationship between the epithelial adhesion molecules and immune responses of airway epithelium, we observed the expression of integrin β4 and intercellular adhesion molecule-1 (ICAM-1) in the mice airway epithelium after sensitization with allergens. BALB/c mice were sensitized with intraperitoneal injection of ovalbumin (OVA) or house dust mite (HDM) and then developed airway hyper-responsiveness as determined by barometric whole-body plethysmography. Both OVA and HDM sensitization led to increases of the number of peripheral leukocytes as well as inflammatory cells infiltration in lungs. OVA sensitized mice showed more severe inflammatory cells infiltration than HDM sensitized mice. Immunohistochemistry analysis of mice lung tissues revealed that sensitization with both allergens also led to a decrease of integrin β4 expression and an increase of ICAM-1 expression in airway epithelia. OVA sensitized mice showed a more significant increase of ICAM-1 expression compared with HDM sensitized mice. siRNA mediated silencing of integrin β4 gene in 16HBE cells resulted in an up-regulation of ICAM-1 expression. Our results indicate a possible role of airway epithelial adhesion molecules in allergen-induced airway immune responses.
Allergens ; pharmacology ; Animals ; Integrin beta4 ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lung ; metabolism ; physiopathology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; Pyroglyphidae ; Respiratory Hypersensitivity ; metabolism

OBJECTIVETo detect the expression and explore the role of the innate immune NLRP3 inflammasome and its downstream factors interleukin-1β (IL-1β)/ interleukin-18 (IL-18) in rat model of allergic rhinitis (AR).

METHODSForty Sprague Dawley (SD) rats were randomly divided into control group (A group), AR model group 1 (B group), AR model group 2(C group), AR model group 3 (D group). Every group contained 10 rats. After the rats in the model group were sensitized by ovalbumin (OVA) and alum, B, C and D groups were separately stimulated with 5% OVA for 10 days, 20 days and 30 days (once a day). The control group did not add OVA in the process of sensitization and excitation. All rats were executed after excitation.Eosinophil granulocyte (EOS) infiltration were observed in nasal mucosa by hematoxylin-eosin (HE) staining, the expression of NLRP3 and cysteinyl aspartate-specific protease-1 (Caspase-1) were observed in nasal mucosa by immunohistochemical staining. The concentrations of ovalbumin specific IgE (OVA-sIgE), IL-18 and IL-1β in peripheral blood and the concentrations of IL-18 and IL-1β in nasal fluid were tested by enzyme-linked immunosorbent assay (ELISA). The data were processed by SPSS 17.0 software.

RESULTSEOS cell counted, the behavioral score and the concentrations of OVA-sIgE in AR model group were obviously higher than those in control group (P < 0.05), and the difference of which had statistical significance between the AR model groups (P < 0.05). The expression of NLRP3 in AR model group (The expression of NLRP3 in group of B, C and D were 48.80 ± 10.75, 71.80 ± 16.98 and 100.32 ± 13.91, respectively) were obviously higher than those in control group (17.47 ± 5.59), the difference of which had statistical significance (F = 78.399, P < 0.05). The expression of Caspase-1 in AR model group (The expression of Caspase-1 in group of B, C and D were 36.33 ± 4.71, 50.87 ± 11.18 and 73.10 ± 14.77, respectively) were obviously higher than those in control group (11.48 ± 2.70), the difference of which had statistical significance (F = 71.727, P < 0.05). The concentrations of IL-1β in AR model group [The concentrations of IL-1β in group of B, C and D were (56.46 ± 10.13), (82.37 ± 11.93), (112.01 ± 22.91) pg/ml, respectively] were obviously higher than those in control group [(38.26 ± 4.66) pg/ml], the difference of which had statistical significance (F = 51.981, P < 0.05). The concentrations of IL-18 in AR model group [The concentrations of IL-18 in group of B, C and D were (177.92 ± 23.63), (194.33 ± 20.78), (234.06 ± 31.70) pg/ml, respectively] were obviously higher than those in control group [(89.71 ± 5.56) pg/ml], the difference of which had statistical significance (F = 73.295, P < 0.05). And the difference of which had statistical significance between the AR model groups (P < 0.05). The expression of NLRP3 was significantly positively correlated with the behavioral score, the concentrations of OVA-sIgE and EOS cell counted in rat model of allergic rhinitis (r value were 0.833,0.873 and 0.868, respectively, all P < 0.01).

CONCLUSIONNLRP3 inflammasome and its downstream factors IL-1β/IL-18 play a role in the pathogenesis of allergic rhinitis, which may be correlated with the degree of inflammation.

Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Inflammasomes ; metabolism ; Inflammation ; Interleukin-18 ; metabolism ; Interleukin-1beta ; metabolism ; Nasal Mucosa ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic ; metabolism

OBJECTIVE: To investigate the expression of thymic stromal lymphopoietin (TSLP) in the nasal mucosa of mouse with allergic rhinitis. METHOD: Twenty wide type BALB/c mouse were divided into 2 groups randomly. Two groups were included, allergic rhinitis group (group A) and control group (group B). The mouse model of allergic rhinitis was established by ovalbumin (OVA) sensitization and challenge. The expressions of TSLP in the nasal mucosa was determined by realtime quantitative PCR and immunohistochemical method. RESULT: The expression of TSLP in the nasal mucosa of group A was significantly higher than that in group B (P<0.01). CONCLUSION TSLP plays a role in the mouse model of allergic rhinitis.
Animals ; Cytokines ; metabolism ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Ovalbumin ; biosynthesis ; Rhinitis, Allergic, Perennial ; metabolism

OBJECTIVE: To determine the expression of nerve growth factor (NGF), tyrosine kinase receptor A (trkA), and pan-neurotrophin receptor (p75) in the lung tissues in asthmatic rats, and to explore their effects on the airway inflammation. METHODS: Thirty-two SD rats were randomly divided into 4 groups: the control, asthma, NGF and anti-NGF groups. The asthmatic model was established by the inhalation and injection of ovalbumin. The total cell count and differential cell count in the bronchoalveolar lavage fluid (BALF) were performed. The pathologic changes in the lung tissues of the 4 groups was detected by HE staining. The NGF mRNA expression in the lung tissues of the asthma and control groups was determined by reverse transcription-polymerase chain reaction (RT-PCR). The changes of trkA and p75 mRNA expressions in the lung tissues in the 4 groups were also investigated by RT-PCR. RESULTS: Compared with the control group, the BALF total cell, the BALF eosinophils (Eos), and the BALF lymphocytes (Lyms) significantly increased (All P <0. 001) in the asthma group; and the lung tissues of the asthma group had more infiltrating inflammatory cells. Not only the expression of NGF mRNA, but also its receptors trkA and p75 mRNA in the lung tissues were significantly higher in the asthma group than those in the control group (All P < 0.01). Positive correlation was found between the expression of NGF mRNA and the BALF total cell, the BALF Lyms in the asthma group. Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the NGF group significantly increased (All P < 0.01), and the lungs of the NGF group had apparent inflammatory changes. The expre-ssions of p75 and trkA mRNA were enhanced significantly (All P < 0.05). Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the anti-NGF group significantly decreased (All P < 0.001), and the lungs of the anti-NGF group showed alleviative inflammatory changes. The expre-ssions of p75 and trkA mRNA significantly decreased (All P < 0.01). CONCLUSION In lungs of asthmatic rats, the elevated expression of NGF mRNA is closely related to the airway inflammation. NGF can upregulate the expressions of p75 and trkA mRNA in asthmatic rats, and then may promote their role in the airway neuronal inflammation in asthma.
Animals ; Asthma ; chemically induced ; metabolism ; Bronchitis ; metabolism ; Lung ; metabolism ; Male ; Nerve Growth Factor ; metabolism ; Ovalbumin ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, trkA ; metabolism ; Receptors, Nerve Growth Factor ; metabolism

Objective: To investigate the effects of dopamine on olfactory function and inflammatory injury of olfactory bulb in mice with allergic rhinitis (AR). Methods: AR mouse model was established by using ovalbumin (OVA), and the mice were divided into two groups: olfactory dysfunction (OD) group and without OD group through buried food pellet test (BFPT). The OD mice were randomly divided into 2 groups, and OVA combined with dopamine (3, 6, 9 and 12 days, respectively) or OVA combined with an equal amount of PBS (the same treatment time) was administered nasally. The olfactory function of mice was evaluated by BFPT. The number of eosinophils and goblet cells in the nasal mucosa were detected by HE and PAS staining. Western blotting, immunohistochemistry or immunofluorescence were used to detect the expression of olfactory marker protein (OMP) in olfactory epithelium, the important rate-limiting enzyme tyrosine hydroxylase (TH) of dopamine, and the marker proteins glial fibrillary acidic protein (GFAP) and CD11b of glial cell in the olfactory bulb. TUNEL staining was used to detect the damage of the olfactory bulb. SPSS 26.0 software was used for statistical analysis. Results: AR mice with OD had AR pathological characteristics. Compared with AR mice without OD, the expression of OMP in olfactory epithelium of AR mice with OD was reduced (F=26.09, P<0.05), the expression of GFAP and CD11b in the olfactory bulb was increased (F value was 38.95 and 71.71, respectively, both P<0.05), and the expression of TH in the olfactory bulb was decreased (F=77.00, P<0.05). Nasal administration of dopamine could shorten the time of food globule detection in mice to a certain extent, down-regulate the expression of GFAP and CD11b in the olfactory bulb (F value was 6.55 and 46.11, respectively, both P<0.05), and reduce the number of apoptotic cells in the olfactory bulb (F=25.64, P<0.05). But dopamine had no significant effect on the number of eosinophils and goblet cells in nasal mucosa (F value was 36.26 and 19.38, respectively, both P>0.05), and had no significant effect on the expression of OMP in the olfactory epithelium (F=55.27, P>0.05). Conclusion: Dopamine can improve olfactory function in mice with AR to a certain extent, possibly because of inhibiting the activation of glial cells in olfactory bulb and reducing the apoptotic injury of olfactory bulb cells.
Animals ; Disease Models, Animal ; Dopamine ; Humans ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa/metabolism* ; Olfactory Bulb/pathology* ; Ovalbumin ; Rhinitis, Allergic/metabolism*

OBJECTIVETo investigate the effects of fulvotomentoside (Ful) on inflammatory factors and antiinflammatory factors in intestine of ovalbumin (OVA)-sensitized BALB/c mice, and to explore the mechanisms of its anti-food allergy effect.

METHODTwenty-four female BALB/c mice aged 6 weeks fed with ovalbumin-free feed were randomly divided into 3 groups, food allergy (FA) group, Ful group and normal saline (NS) group. Mice in FA and Ful groups were sensitized intraperitoneally two times with OVA and challenged intragastrically with OVA. Mice in Ful group were treated with 200 mg/kg of Ful by subcutaneous injection once daily for 22 days. The mice in FA and NS groups were used as positive control and negative control, respectively, and were treated with normal saline solution by subcutaneous injection for 22 days. Just 48 hours after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. The mRNA expressions of transforming growth factor β1 (TGF-β1), interleukin-6 (IL-6), interleukin-17A (IL-17A) and forkhead box P3 (Foxp3) in jejunum were detected by reverse transcription-PCR (RT-PCR). The protein expressions of TGF-β1, IL-6, and IL-17A in jejunum were detected by immunohistochemical method. The activation of neutrophils in jejunum was assayed by the levels of MPO.

RESULTThe expressions of TGF-β1, IL-6, IL-17A mRNA [(0.370 ± 0.013), (0.475 ± 0.015), (0.541 ± 0.013)] and related protein [(53,075.70 ± 20,727.06), (256,881.66 ± 36,561.79), (435,064.25 ± 69,911.48)] in jejunum were increased and the Foxp3 mRNA [(0.231 ± 0.014) vs. (0.365 ± 0.015)] expression was decreased in group FA. After the treatment with Ful, IL-6 and IL-17A mRNA [(0.196 ± 0.005), (0.204 ± 0.008)] and protein [(114,040.30 ± 20,295.25), (218,200.74 ± 30,077.69)] expressions were decreased and Foxp3 mRNA (0.578 ± 0.021) expression was increased, and no change of TGF-β1 expression was unchanged. There were no significant differences of the levels of MPO among the three groups.

CONCLUSIONInflammatory reaction which was characterized by the increase of IL-6 and IL-17A expressions was found in intestine of ovalbumin-sensitized BALB/c mice. Ful could decrease overexpression of IL-6 and IL-17A, and increase the expression of specific transcription factor Foxp3 of regulatory T cells significantly in intestine. It may be one of the mechanisms that Ful improved intestinal inflammation.

Animals ; Female ; Food Hypersensitivity ; metabolism ; Forkhead Transcription Factors ; metabolism ; Inflammation ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Intestines ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Ovalbumin ; adverse effects ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo explore the effect of food allergy (FA) on the development of visceralgia sensibility and the substance P (SP) expression in colon of developing rats with FA.

METHODThree-week old female Sprague-Dawley (SD) rats were randomly divided into two groups (n = 10 in each). The rats in FA group were sensitized with ovalbumin (OVA) 40 µg and Al(OH)3 1 mg suspension solution (0.2 ml) intraperitoneal (i.p.) injection on day 0, only OVA 40 µg solution i.p. on day 2, 4, 7, 9, 11, respectively, and the rats were challenged by gavage with OVA solution 30 mg (2 ml) on day 20, 24, 28, 30. The rats in non-sensitized (NS) group were not challenged except handled in the same ways. The serum OVA-IgE were determined by enzyme-linked immuno sorbent assay (ELISA) on day 0, 30. Jejunum segments were used to observe morphological structure, the expression of eosinophils, and the density and the percentage of degranulation of mast cells (MC). The rats were appraised for the pain sensibility of intestinal tract under colorectal distension irritation by the electrophysiological method on external oblique in the 18-24 hr after the last challenge. Colons were used to analyze the expression of SP through immunohistochemical staining and computer image analyzing system.

RESULTThe serum OVA-IgE concentration and the eosinophils, mast cell, the percentage of mast cells degranulation in FA group were more than NS group (P < 0.01). The amplitudes of spike external oblique muscle of abdomen (EOMA, µV) of the FA group under the colorectal distension (CRD) pressures at 0, 15, 30, 45, 60, 75 mm Hg were (17.74 ± 0.72), (18.63 ± 1.72), (22.55 ± 1.70), (28.63 ± 7.00), (33.97 ± 7.34), (37.26 ± 8.40), and (17.43 ± 1.18), (17.27 ± 1.16), (17.73 ± 1.42), (19.55 ± 3.54), (23.29 ± 5.46), (25.20 ± 4.75) in NS group. With the CRD pressure increased, the amplitudes of spike EOMA increased significantly. There were significant differences between groups under the CRD pressures at 30, 45, 60, 75 mm Hg (F = 47.470, 13.367, 13.317, 15.390, P < 0.01). The expressions of colons SP in FA group and NS group are 247.12 ± 90.83 and 103.90 ± 58.94, respectively (t = 4.183, P < 0.01).

CONCLUSIONSensitization through i.p. pathway and challenge by gavage with OVA in early life could result in FA in young SD rats. FA in early life enabled the amplitudes of spike EOMA and the expression of colons SP increase significantly. It may be related to increase in amount and degranulation of MC and SP abnormal expression in colon, which could lead to the development of visceralgia sensibility.

Animals ; Colonic Diseases, Functional ; metabolism ; Disease Models, Animal ; Electrophysiology ; Female ; Food Hypersensitivity ; complications ; metabolism ; Hyperalgesia ; etiology ; metabolism ; physiopathology ; Intestinal Mucosa ; metabolism ; pathology ; Mast Cells ; metabolism ; Ovalbumin ; adverse effects ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; Substance P ; metabolism

OBJECTIVETo explore the role of transient receptor potential canonical 1 (TRPC1) in airway remodeling and the effect of budesonide intervention on its expression in the lungs of guinea pigs with ovalbumin-induced asthma.

METHODSFifty male guinea pigs were randomized into 5 equal groups, including a blank control group, ovalbumin group, ovalbumin+TRPC1 siRNA group, ovalbumin+luciferase siRNA group, and ovalbumin+budesonide group. After corresponding treatments, bronchoalveolar lavage was collected from the guinea pigs for eosinophils analysis and detection of IL-5 and IL-13 levels using ELISA. The lung tissues were stained with HE and Masson's trichrome to observe the bronchial wall thickness, smooth muscle hypertrophy, subepithelial collagen deposition, and lung inflammations. Immunohistochemistry and real-time quantitative PCR were performed to detect TRPC1 protein and mRNA expressions in the lungs, respectively.

RESULTSThe guinea pig models of ovalbumin-induced asthma showed significantly increased thickness of the bronchial wall, smooth muscle hypertrophy, collagen deposition and inflammatory cell infiltration, but these pathologies were obviously alleviated by treatment with TRPC1 siRNA or budesonide (P/0.05). Immunohistochemstry showed that TRPC1 protein was distributed mainly on the cell membrane and in the nuclei of the basal cells or columnar epithelial cells.

CONCLUSIONThe up-regulated expression of TRPC1 ion channel is closely associated with the occurrence and progression of airway remodeling and chronic airway inflammation in asthma. Budesonide can partially suppress airway remodeling and inflammation by regulating the expression of TRPC1.

Airway Remodeling ; Animals ; Asthma ; drug therapy ; metabolism ; Bronchi ; pathology ; Budesonide ; pharmacology ; Disease Models, Animal ; Guinea Pigs ; Inflammation ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-5 ; metabolism ; Leukocyte Count ; Lung ; drug effects ; metabolism ; Male ; Ovalbumin ; TRPC Cation Channels ; metabolism