This study aimed to investigate the mechanism by which Shegan Mahuang Decoction(SGMH) and its bitter Chinese herbs(BCHs) regulated the lung-gut axis through the bitter taste receptor 14(TAS2R14)/secretory immunoglobulin A(SIgA)/thymic stromal lymphopoietin(TSLP) to intervene in the epithelial cell barrier of cold asthma rats. Fifty SD rats were randomly divided into the following five groups: normal group, model group, dexamethasone group, SGMH group, and BCHs group. A 10% ovalbumin(OVA) solution was used to sensitize the rats via subcutaneous injection on both sides of the abdomen and groin, combined with 2% OVA atomization and cold(2-4 ℃) stimulation to induce a cold asthma model in rats. The SGMH, BCHs, and dexamethasone groups were given corresponding treatments by gavage and nebulization, while the normal and model groups received normal saline by gavage and nebulization. After the final stimulation, pathological changes in the lung and intestine tissues were observed using hematoxylin-eosin(HE) and periodic acid-Schiff(PAS) staining. Lung function was assessed by measuring the ratio of forced expiratory volume in the first second to forced vital capacity(FEV1/FVC), the ratio of the average flow rate at 25%-75% of forced vital capacity to foned vital capacity(FEV25%-75%/FVC), the peak expiratory flow(PEF), and pulmonary resistance(RL). The levels of IL-4, IL-5, IL-13, and TNF-α in serum, and sIgA in serum, intestinal, and bronchial mucosa were detected by enzyme-linked immunosorbent assay(ELISA). The expression of TAS2R14 protein in lung tissue was detected by Western blot(WB). The content of short-chain fatty acids(SCFAs) in rat feces was determined by gas chromatography-mass spectrometry(GC-MS). The effect of TAS2R14/TSLP on lipopolysaccharide(LPS)-induced inflammation in epithelial cells in the BCHs group was observed, and the expression of TAS2R14 and TSLP in cells was detected by WB. Compared with the normal group, the model group showed reduced water intake, diet, and body weight, increased infiltration of inflammatory cells in the lung and intestinal tissues, goblet cell hyperplasia, significantly decreased FEV1/FVC, FEV25%-75%/FVC, and PEF, and significantly increased RL. Moreover, serum levels of IL-4, IL-5, IL-13, and TNF-α were elevated, and sIgA levels in serum, intestine, and bronchial mucosa were significantly decreased. TAS2R14 expression in lung tissues was inhibited, and the content of acetic acid, propionic acid, and butyric acid in feces was significantly reduced. In the LPS group, TSLP expression increased, and TAS2R14 expression decreased. Compared with the model group, the general condition of rats in the SGMH and BCHs groups improved, with reduced infiltration of inflammatory cells and goblet cell hyperplasia in the lung and intestinal tissues. FEV1/FVC, FEV25%-75%/FVC, and PEF significantly increased, and RL significantly decreased. Serum levels of IL-4, IL-5, IL-13, and TNF-α decreased, while sIgA levels in serum, intestine, and bronchial mucosa significantly increased, and TAS2R14 expression was activated in lung and intestinal tissues. The content of acetic acid, propionic acid, and butyric acid in feces significantly increased. Compared with the model group, the BCHs group and the agonist group showed inhibited TSLP expression and increased TAS2R14 expression. The results showed that both SGMH and BCHs could reduce lung and intestinal inflammatory reactions, improve lung function, and regulate the content of intestinal SCFAs in asthmatic rats. There was no significant difference in TAS2R14 protein expression between the SGMH and BCHs groups, indicating that the clinical efficacy of BCHs may be related to the activation of the bitter receptor TAS2R14 and the regulation of immune inflammatory mediators in lung and intestinal epithelial cells.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Rats, Sprague-Dawley ; Lung/metabolism* ; Asthma/metabolism* ; Cytokines/immunology* ; Male ; Receptors, G-Protein-Coupled/immunology* ; Epithelial Cells/metabolism* ; Thymic Stromal Lymphopoietin ; Immunoglobulin A, Secretory/genetics* ; Humans ; Cold Temperature

OBJECTIVES: To investigate the protective effect of the probiotic bacterium Streptococcus salivarius K12 (K12) against Mycoplasma pneumoniae (Mp) infection in mice. METHODS: Forty male BALB/c mice were randomized into normal control group, K12 treatment group, Mp infection group, and K12 pretreatment prior to Mp infection group. The probiotic K12 was administered daily by gavage for 14 days before Mp infection induced by intranasal instillation of Mp. Three days after Mp infection, the mice were euthanized for analysis of bronchoalveolar lavage fluid (BALF) cell counts and serum levels of secretory immunoglobulin A (sIgA), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6). RT-qPCR was performed to detect the P1 and community-acquired respiratory distress syndrome ( CARDS ) toxin of Mp in the lung tissues and the mRNA expressions of TNF-α, IL-6, chemokine 1 (CXCL1), matrix metalloproteinase 9 (MMP9), mucin 5ac (MUC5ac), collagen 3a1 (Col3a1), Toll-like receptor 2 (TLR2) and TLR4; the protein expressions of TLR2 and TLR4 in the lung tissue were detected using Western blotting. Pathological changes in the lung tissue and airway remodeling were examined with HE staining and AB/PAS staining. RESULTS: Compared with the Mp-infected mice with PBS treatment, the infected mice with K12 treatment showed significantly lowered mRNA levels of P1 and CARDS in the lung tissue and reduced white blood cell counts in the BALF (P<0.05). In spite of the absence of significant differences in serum levels of inflammatory factors between the two groups, the mRNA expressions of TNF‑α, IL-6, CXCL1, MMP9, MUC5ac and COL3A1 and the mRNA and protein levels of TLR2 and TLR4 in the lung tissues were significantly lower in K12-treated mice, in which AB/PAS staining showed obviously decreased mucus secretion. CONCLUSIONS K12 pretreatment can effectively reduce pulmonary inflammatory responses, improve airway remodeling and alleviate lung injury in Mp-infected mice.
Animals ; Mice ; Pneumonia, Mycoplasma/metabolism* ; Mice, Inbred BALB C ; Toll-Like Receptor 2/metabolism* ; Mycoplasma pneumoniae ; Male ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Lung/microbiology* ; Toll-Like Receptor 4/metabolism* ; Streptococcus salivarius ; Probiotics/administration & dosage* ; Bronchoalveolar Lavage Fluid ; Matrix Metalloproteinase 9/metabolism* ; Mucin 5AC/metabolism* ; Chemokine CXCL1/metabolism* ; Immunoglobulin A, Secretory/metabolism* ; Bacterial Toxins ; Bacterial Proteins

The effects of diclazuril on the bursa of Fabricius (BF) structure and secretory IgA (SIgA) expression in chickens infected with Eimeria tenella were examined. The morphology of the BF was observed by hematoxylin and eosin staining, while ultrastructural changes were monitored by transmission electron microscopy. E. tenella infection caused the BF cell volumes to decrease, irregularly arranged, as well as, enlargement of the intercellular space. Diclazuril treatment alleviated the physical signs of damages associated with E. tenella infection. The SIgA expression in BF was analyzed by immunohistochemistry technique. The SIgA expression increased significantly by 350.4% (P<0.01) after E. tenella infection compared to the normal control group. With the treatment of diclazuril, the SIgA was relatively fewer in the cortex, and the expression level was significantly decreased by 46.7% (P<0.01) compared with the infected and untreated group. In conclusion, E. tenella infection in chickens induced obvious harmful changes in BF morphological structure and stimulated the expression of SIgA in the BF. Diclazuril treatment effectively alleviated the morphological changes. This result demonstrates a method to develop an immunological strategy in coccidiosis control.
Animals ; Bursa of Fabricius/anatomy & histology/*parasitology ; Chickens ; Coccidiosis/drug therapy/metabolism/parasitology/*veterinary ; Coccidiostats/administration & dosage/*adverse effects ; Eimeria tenella/*physiology ; Female ; Immunoglobulin A, Secretory/*genetics/metabolism ; Male ; Nitriles/administration & dosage/*adverse effects ; Poultry Diseases/*drug therapy/genetics/metabolism/parasitology ; Triazines/administration & dosage/*adverse effects

OBJECTIVETo explore the expressions of SIgA and alpha l-AR in benign prostatic hyperplasia (BPH) complicated by chronic prostatitis (CP) and their implications.

METHODSAccording to the preoperative findings of expressed prostatic secretion (EPS), transrectal prostate ultrasonography, prostate-specific antigen (PSA), international prostate symptom score (IPSS), clinical symptoms, chronic pelvic pain syndrome (CPPS) and postoperative histopathology, 62 cases of BPH pathologically confirmed after transurethral plasma kinetic resection of the prostate (PKRP) were divided into a BPH group (n = 32) and a BPH + CP group (n = 30). The expressions of SIgA and alpha 1-AR in the prostate tissue were determined by immunohistochemistry and PT-PCR.

RESULTSOf the 62 cases, 30 were found to be BPH + CP, and the other 32 to be BPH. The expressions of SIgA and alpha1-AR were significantly higher in the BPH + CP than in the BPH group (0.380 8 +/- 0.144 3 vs 0.295 4 +/- 0.008 4 and 0.440 5 +/- 0.104 1 vs 0.383 2 +/- 0.013 6, P < 0.05).

CONCLUSIONThe upregulated expressions of SIgA and alpha1-AR expression in BPH complicated by CP suggest a certain association between CP and BPH, and that inflammation may be a pathogenic factor of BPH and correlate with its pathological development.

Aged ; Chronic Disease ; Humans ; Immunoglobulin A, Secretory ; metabolism ; Male ; Middle Aged ; Prostate ; metabolism ; pathology ; Prostatic Hyperplasia ; complications ; metabolism ; pathology ; Prostatitis ; complications ; metabolism ; pathology ; Receptors, Androgen ; metabolism

OBJECTIVETo investigate the effect of total alkaloids of Sophora alopecuroides (TASA) on dextran sulfate sodium (DSS)-induced colitis in mice.

METHODSChronic experimental colitis was induced by administration of 4 cycles of 4% DSS. Fifty mice were randomly distributed into 4 groups (normal, DSS, DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification. Mice in the normal group (n=11) and DSS-induced colitis control group (n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups (n=12 each) were treated with TASA solution (20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg, respectively. The severity of colitis was assessed on the basis of clinical signs, colon length, and histology scores. Moreover, secretory immunoglobulin A (sIgA) and haptoglobin (HP) were analyzed by enzyme linked immunosorbent assay; intercellular adhesion molecule 1 (ICAM-1) and macrophage-migration inhibitory factor (MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction (qRT-PCR) using SYBA green I; and nuclear factor κ B (NF-κ B) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay.

RESULTSTASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum sIgA. TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression. Also, TASA was able to reduce phospho-I κ B α (p-I κ B α) protein expression; however, it had no effect on the activation of I κ B kinase α (IKK α) and inhibitor of NF-κ B α (I κ B α). Moreover, TASA inhibited the p65 recruitment to the ICAM-1 gene promoter.

CONCLUSIONSTASA had a protective effect on DSS-induced colitis. Such effect may be associated with its inhibition of NF-κ B activation and blockade of NF-κ B-regulated transcription activation of proinflammatory mediator gene.

Alkaloids ; pharmacology ; therapeutic use ; Animals ; Cecum ; drug effects ; metabolism ; pathology ; Colitis ; chemically induced ; drug therapy ; pathology ; prevention & control ; Colon ; pathology ; ultrastructure ; Dextran Sulfate ; Down-Regulation ; drug effects ; Female ; Haptoglobins ; metabolism ; I-kappa B Proteins ; metabolism ; Immunoglobulin A, Secretory ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Intestinal Mucosa ; drug effects ; pathology ; ultrastructure ; Macrophage Migration-Inhibitory Factors ; genetics ; metabolism ; Mice ; Mice, Inbred C57BL ; NF-KappaB Inhibitor alpha ; Phosphorylation ; drug effects ; Phytotherapy ; Promoter Regions, Genetic ; genetics ; Protective Agents ; pharmacology ; therapeutic use ; Protein Binding ; drug effects ; Signal Transduction ; drug effects ; Sophora ; chemistry ; Transcription Factor RelA ; metabolism

Animals ; Immunoglobulin A, Secretory ; metabolism ; Intestines ; drug effects ; metabolism ; Male ; Massive Hepatic Necrosis ; metabolism ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the influence of early enteral immunonutrition on immunological function of body and intestine in severely scalded rats.

METHODSSixty-four healthy adult SD rats were subjected to 30% TBSA full-thickness scald and randomly divided into enteral nutrition group (EN, n = 32) and enteral immunonutrition group (EIN, n = 32). Another 8 healthy rats were enrolled as normal controls. The changes in CD3+, CD4+, CD8+ T lymphocyte subsets, the PCNA expression in intestinal mucosa,the amount of plasmocytes in lamina propria of small intestine and the content of s-IgA in small intestine were determined on 1, 4, 7 and 10 post-burn days (PBD).

RESULTS(1) CD3+, CD4+ and the ratio of CD4+/CD8+ were remarkably declined, while CD8+ was obviously raised after scald in EN group, and there were statistically significant differences at most of time-points when compared with those in control group (P < 0.05 or P < 0.01). CD3+, CD4+ and the ratio of CD4+/CD8+ on 7 and 10 PBD in EIN group were obviously higher (increased from 4 PBD on, and it was 1.86 +/- 0.25 on 10 PBD) than those in EN group (CD4+/CD8+ was 1.26 +/- 0.10 on 10 PBD), while CD8+ in EIN group was evidently lower than that in EN group (P < 0.01). (2) The level of PCNA expression in intestinal mucosa, the amount of plasmocytes in lamina propria of small intestine and the content of s-IgA in small intestine in EIN group were increased significantly compared with EN group on 4, 7, 10 PBD (P < 0.05 or P < 0.01).

CONCLUSIONEarly enteral immunonutrition can improve immunological function of body and intestine after severe scald, with better results than enteral nutrition.

Animals ; Arginine ; Burns ; immunology ; therapy ; CD4-CD8 Ratio ; Enteral Nutrition ; Glutamine ; Immunoglobulin A, Secretory ; metabolism ; Intestinal Mucosa ; metabolism ; Intestine, Small ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

AIMTo study the effect of the recombined human growth hormone(rhGH) on secretory immunoglobulin A (sIgA), epidermal growth factor (EGF) in rats with obstructive jaundice.

METHODSSixty Wistar rats were randomly divided into four groups, sham-operated (group A), common biliary duct-ligated (group B), biliary duct-ligated plus rhGH-treat for one week (group C), biliary duct-ligated plus rhGH-treat for two weeks (group D), each group had 15 rats. Except group A, the rats of other groups were operated with biliary duct-ligated. Until two weeks after operation, the rats of group A and B were killed. After operation, the rats of group C were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for one week, and then killed. The rats of D group were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for two weeks, and then killed. All procedures were performed aseptically. Total bilirubin (TB), alkaline phosphatase (ALP), prealbumin(PA), insulin-like growth factor (IGF-1), sIgA, EGF were measured.

RESULTSCompared with group A, in group B, C, D, serum level of PA, IGF-1 and sIgA, EGF level of gastric and intestinal juice were lower, but TB, ALP were higher, there were significant difference. Compared with group B, the rats with treatment of rhGH in group C and D had higher sIgA and EGF and lower intestinal bacterial translocation.

CONCLUSIONIn objective jaundice rats, rhGH can protect their hepatic function, intestinal physical-barrier function and immune-barrier function, and reduce intestinal bacterial translocation.

Animals ; Bacterial Translocation ; Epidermal Growth Factor ; metabolism ; Growth Hormone ; pharmacology ; Humans ; Immunoglobulin A, Secretory ; metabolism ; Jaundice, Obstructive ; metabolism ; Rats ; Rats, Wistar ; Recombinant Proteins ; pharmacology

OBJECTIVETo valuate the intestinal mucosal secretary IgA (sIgA) responses to the ovalbumin-induced allergy in mice, to provide some clues for the exploration of mechanisms and therapeutic methods in the children's food allergy.

METHODSFemale BALB/c mice aged 6 weeks fed on the ovalbulmin-free diet, were randomly divided into 2 groups with 8 mice in each. The mice in group Ch were sensitized with ovalbumin (OVA) intraperitoneal injection two times and challenged intragastrically 3 times. Two days after the last challenge with oral OVA, the mice were sacrificed and the samples were collected. The mice in group Ns were given intraperitoneal and intragastrical normal saline as control. The levels of total IgA and OVA-specific IgA in the intestinal mucus of the mice were determined by ELISA; the immunohistochemical methods were adopted to observe IgA(+) plasmacytes in lamina propria (LP) and surface membrane IgA (smIgA)(+) lymphocytes in peyer's patch (PP); the IL-4 mRNA expression in LP was assessed by RT-PCR. The IL-4 mRNA expression in PP was evaluated by in situ hybridization.

RESULTSAfter the mice in Ch group were sensitized and challenged with OVA, the levels of the total IgA and the OVA-specific IgA in mucus remarkably increased (P < 0.01 respectively), the amounts of the IgA(+) plasmacytes in LP and the smIgA(+) lymphocytes in PP significantly increased (P < 0.01 respectively); a significantly positive correlation was found among the total IgA levels, the OVA-specific IgA levels, the IgA(+) plasmacyte counts in LP and the smIgA(+) lymphocyte counts in PP (P < 0.01 respectively); the mRNA expressions of IL-4 in LP and in PP were significantly augmented (P < 0.01 respectively); significantly positive correlations were found either between the IL-4 mRNA expression and the IgA(+) plasmacyte counts in LP (P < 0.01) or between the IL-4 mRNA expression and the smIgA(+) lymphocyte counts in PP (P < 0.01).

CONCLUSIONSThe intestinal mucosal sIgA responses are abnormally augmented in the ovalbumine-induced allergic mice, which may be partly due to the increased expression of IL-4 mRNA in gut.

Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Female ; Food Hypersensitivity ; immunology ; Immunoglobulin A, Secretory ; metabolism ; Immunohistochemistry ; Interleukin-4 ; genetics ; metabolism ; Intestinal Mucosa ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; administration & dosage ; immunology ; Peyer's Patches ; immunology ; RNA, Messenger ; metabolism ; Random Allocation

OBJECTIVETo test the salivary immunoglobulin A antibody activity to Streptococcus mutans in normal with in acid environment.

METHODSStreptococcus mutans strains were isolated from 20 volunteers, serotyped by biochemical test and PCR, and genotyped by AP-PCR. Unstimulated secretions from submandibular glands and sublingual glands were collected from volunteers by modified collectors. Each identified Streptococcus mutans genotype was cultured in two groups: control group was cultured in BHI broth pH7.2 at 37 degrees C for 2 h; acid shock group were cultured in TYEG broth (pH5.5) at 37 degrees C for 2 h. Analysis of SIgA activity to Streptococcus mutans genotypes in different groups was detected by Western blot.

RESULTS(1) The SIgA of each individual could response to his own Streptococcus mutans strains and the reference strains; (2) The same individual had different SIgA activity to different genotype strains; (3) There were no significant difference between acid groups and control groups, in spite that some bands had strong or weak intensity.

CONCLUSIONSAlthough Streptococcus mutans could express acid shock proteins in stress, the present study suggests that these new proteins have no qualitative effect on the reaction of SIgA to Streptococcus mutans.

Adult ; Dental Plaque ; immunology ; Female ; Humans ; Hydrogen-Ion Concentration ; Immunoglobulin A, Secretory ; immunology ; In Vitro Techniques ; Male ; Middle Aged ; Saliva ; immunology ; Streptococcus mutans ; immunology ; metabolism