Idiopathic interstitial pneumonia (IIP) is characterized by varying degrees of interstitial fibrosis. IL-13 and IL-4 are strong inducers of tissue fibrosis, whereas IFN-gamma has antifibrotic potential. However, the roles of these substances in IIP remain unknown. IL-13, IL-4, and IFN-gamma were measured in the BAL fluid of 16 idiopathic pulmonary fibrosis (IPF) patients, 10 nonspecific interstitial pneumonia (NSIP) patients, and 8 normal controls. The expression of IL-13 and IL-13Ralpha1/alpha2 in lung tissues was analyzed using ELISA and immunohistochemistry. IL-13 levels were significantly higher in IPF patients than the others (P<0.05). IL-4 levels were higher in both IPF and NSIP patients than in normal controls (P<0.05), and IFN-gamma levels were lower in NSIP patients than in normal controls (P=0.047). IL-13 levels correlated inversely with FVC% (r=-0.47, P=0.043) and DLCO% (r=-0.58, P=0.014) in IPF and NSIP patients. IL-13 was strongly expressed in the smooth muscle, bronchial epithelium, alveolar macrophages and endothelium of IPF patients. IL-13Ralpha1, rather than IL-13Ralpha2, was strongly expressed in the smooth muscle, bronchial epithelium, and endothelium of IPF patients. IL-13 and its receptors may contribute to the pathogenesis of fibrosis in IIP and appear to be related to the severity of the disease.
Adult ; Female ; Humans ; Idiopathic Interstitial Pneumonias/diagnosis/*metabolism ; Idiopathic Pulmonary Fibrosis/diagnosis/*metabolism ; Interferon-gamma/analysis ; Interleukin-13/*analysis ; Interleukin-13 Receptor alpha1 Subunit/*metabolism ; Interleukin-13 Receptor alpha2 Subunit/*metabolism ; Interleukin-4/analysis ; Lung/physiopathology ; Male ; Middle Aged

OBJECTIVETo evaluate the expressions of interleukin (IL) 13 and its receptor IL-13Rα2 in eosinophilic chronic rhinosinusitis with nasal polyps (ECRSwNP) and non-ECRSwNP and their clinicopathological implications.

METHODSA total of 60 consecutive patients with CRSwNP who underwent endoscopic sinus surgery (ESS) were divided into two groups of ECRSwNP (n = 27) and non-ECRSwNP (n = 33) based on tissue eosinophil count (more than five cells per high power field) with postoperative pathological examination. Before ESS,the severities of symptoms, nasal polyps, and sinonasal diseases on CT were scored, peripheral blood eosinophil count and percentage, and total serum. IgE level were measured. IL-13 and IL-13Rα2 expressions in polyp tissues were examined with immunohistochemistry. SPSS 16.0 software was used to analyze the data.

RESULTSThere was no significant differences between ECRSwNP and non-ECRSwNP groups in the mean symptom scores (t = 0.102, P > 0.05), but ECRSwNP, compared to non- ECRSwNP, demonstrated a higher incidence of bilateral polyps (χ2 = 15.879, P < 0.01), a higher mean score of nasal polyps (3.6 ± 1.1 vs 2.1 ± 0.8, t = 4.009, P < 0.01) or diseases on CT (t = 4.428, P < 0.01). And also a significant difference existed between two groups in mean blood eosinophil count (t = 3.148, P < 0.01) or percentage (t =3.038, P < 0.01), but no significant difference in total serum levels of IgE (t = 0.659, P > 0.05). There was a closed correlation between tissue eosinophil count and blood eosinophil count (r = 0.683, P < 0.01) or percentage (r = 0.631, P < 0.01) in ECRSwNP, but not in non-ECRSwNP. The expressions of both IL-13 and IL-13Rα2 increased significantly in ECRSwNP compared to non-ECRSwNP ( scores 8.1 ± 1.6 vs. 5.4 ± 1.6; 8.8 ± 1.4 vs. 4.8 ± 1.6, t value was 4.749, 8.010, P < 0.01).

CONCLUSIONSIL-13 and IL-13Rα2 are associated closely with pathogenesis o ECRSwNP. Subtyping CRSwNP and studying underly mechanism can be helpful to make treatment strategy for CRSwNP.

Chronic Disease ; Eosinophils ; Humans ; Interleukin-13 ; metabolism ; Interleukin-13 Receptor alpha2 Subunit ; metabolism ; Leukocyte Count ; Nasal Polyps ; metabolism ; Paranasal Sinuses ; Rhinitis ; metabolism ; Sinusitis ; metabolism

Immunoglobulin G4-related sialadenitis (IgG4-RS) is an immune-mediated fibro-inflammatory disease and the pathogenesis is still not fully understood. The aim of this study was to explore the role and mechanism of interleukin-13 (IL-13) in the cellular senescence during the progress of IgG4-RS. We found that the expression of IL-13 and IL-13 receptor α1 (IL-13Rα1) as well as the number of senescent cells were significantly higher in the submandibular glands (SMGs) of IgG4-RS patients. IL-13 directly induced senescence as shown by the elevated activity of senescence-associated β-galactosidase (SA-β-gal), the decreased cell proliferation, and the upregulation of senescence markers (p53 and p16) and senescence-associated secretory phenotype (SASP) factors (IL-1β and IL-6) in SMG-C6 cells. Mechanistically, IL-13 increased the level of phosphorylated signal transducer and activator of transcription 6 (p-STAT6) and mitochondrial-reactive oxygen species (mtROS), while decreased the mitochondrial membrane potential, ATP level, and the expression and activity of superoxide dismutase 2 (SOD2). Notably, the IL-13-induced cellular senescence and mitochondrial dysfunction could be inhibited by pretreatment with either STAT6 inhibitor AS1517499 or mitochondria-targeted ROS scavenger MitoTEMPO. Moreover, IL-13 increased the interaction between p-STAT6 and cAMP-response element binding protein (CREB)-binding protein (CBP) and decreased the transcriptional activity of CREB on SOD2. Taken together, our findings revealed a critical role of IL-13 in the induction of salivary gland epithelial cell senescence through the elevated mitochondrial oxidative stress in a STAT6-CREB-SOD2-dependent pathway in IgG4-RS.
Cellular Senescence/genetics* ; Humans ; Immunoglobulin G/metabolism* ; Interleukin-13/pharmacology* ; Mitochondria/metabolism* ; Sialadenitis/metabolism*

OBJECTIVE: To explore the relationship of the expression of thymic stromal lymphopoietin (TSLP) in nasal polyps tissues and the Th2 inflammatory response. METHOD: Sixty patients with nasal polyps were collect ed. The immunohistochemical staining method was used to detect the expression of TSLP in nasal polyps tissues and ELISA method to the expression of IL-4, IL-5, IFN-gamma, IL-13 and analyzed the correlation between them. RESULT: The expression of TSLP in nasal polyp tissues was higher than that in normal inferior turbinate mucosa (P < 0.05). The expression level of IL-4, IL-5, IFN-gamma and IL-13 in nasal polyps tissues were significantly higher than those in control group (P < 0.05). TSLP staining was a statistically significant positive correlation with IL-4, IL5 and IL-13 (r = 0.475, 0.594 and 0.582, respectively, P < 0.01), while inverse correlation with IFN-gamma (r = -0.614, P < 0.01). CONCLUSION The high expression of TSLP might promote T cell differentiation towards Th2, and participated in the occurrence/development of nasal polyps, aggravated the nose Th2 inflammatory response.
Adult ; Cytokines ; metabolism ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-5 ; metabolism ; Male ; Nasal Polyps ; immunology ; metabolism ; Th2 Cells ; immunology ; Young Adult

OBJECTIVE: To explore and verify that transient receptor potential vanilloid 4(TRPV4) affects chondrocyte degeneration. METHODS: Neonatal SD rats were selected, primary chondrocytes were extracted, and identified by toluidine blue staining and alcian blue staining;an in vitro chondrocyte inflammation model was constructed by IL-1β, and TRPV4 inhibitor was used to treat chondrocytes under inflammatory conditions, and the chondrocytes were treated by RT-PCR method was used to detect matrix metallopeptidase 13(MMP-13), a disintegrin and metalloproteinase with thrombospondin 5, (ADAMTS-5)、nitric oxide synthase 2(NOS2)、Collagen, type II alpha 1(Col2α1)and aggrecan (Acan) mRNA in chondrocytes; primary chondrocytes were treated with different concentrations of TRPV4 overexpression plasmid, and the optimal overexpression dose was screened. The mRNA expressions of TRPV4, MMP-13, ADAMTS-5, NOS2, Col2α1 and Acan in chondrocytes under the optimal TRPV4 overexpression dose were detected. RESULTS: Toluidine blue staining and Alcian blue staining identified the extracted cells as primary chondrocytes;RT-PCR showed that TRPV4, MMP-13, ADAMTS-5, NOS2 mRNA in chondrocytes treated with TRPV4 inhibitor under inflammatory conditions. The expression of Col2α1 mRNA was significantly decreased (P<0.05), and the expression of Col2α1 mRNA was increased (P<0.05). Although there was no significant difference in the expression of Acan mRNA, the overall trend was also increasing. The expression of Col2α1 and Acan mRNA in chondrocytes was significantly decreased (P<0.05), and the expression of NOS2 mRNA was increased(P<0.05), but there was no significant difference in MMP-13 and ADAMTS-5 (P>0.05). CONCLUSION Inhibiting the expression of TRPV4 can down-regulate the expression of genes related to chondrocyte degeneration.
Animals ; Rats ; Aggrecans/metabolism* ; Cartilage, Articular ; Cells, Cultured ; Chondrocytes ; Interleukin-1beta/metabolism* ; Matrix Metalloproteinase 13/metabolism* ; Rats, Sprague-Dawley ; RNA, Messenger/metabolism* ; TRPV Cation Channels/metabolism*

BACKGROUNDExpression of murine calcium-activated chloride channel family member 3 (mCLCA3) has been reported to be increased in the airway epithelium of asthmatic mice challenged with ovalbumin (OVA). However, its role in asthmatic airway inflammation under no OVA exposure has not yet been clarified.

METHODSmCLCA3 plasmids were transfected into the airways of normal BALB/c mice. mCLCA3 expression and airway inflammation in mouse lung tissue were evaluated. Cell differentials and cytokines in bronchoalveolar lavage fluid (BALF) were analyzed. The expression of mCLCA3 protein and mucus protein mucin-5 subtype AC (MUC5AC) were analyzed by Western blotting. The mRNA levels of mCLCA3, MUC5AC and interleukin-13 (IL-13) were determined quantitatively.

RESULTSmCLCA3 expression was not detected in the control group while strong immunoreactivity was detected in the OVA and mCLCA3 plasmid groups, and was strictly localized to the airway epithelium. The numbers of inflammatory cells in lung tissue and BALF were increased in both mCLCA3 plasmid and OVA groups. The protein and mRNA levels of mCLCA3 and MUC5AC in the lung tissue were significantly increased in the mCLCA3 plasmid and OVA groups compared to the control group. The level of IL-13, but not IL-4, IL-5, IFN-γ, CCL2, CCL5 or CCL11, was significantly increased compared with control group in BALF in the mCLCA3 plasmid and OVA groups. The level of IL-13 in the BALF in the mCLCA3 plasmid group was much higher than that in the OVA group (P < 0.05). The level of mCLCA3 mRNA in lung tissue was positively correlated with the levels of MUC5AC mRNA in lung tissue, IL-13 mRNA in lung tissue, the number of eosinophils in BALF, and the content of IL-13 protein in BALF. The level of IL-13 mRNA in lung tissue was positively correlated with the number of eosinophils in BALF and the level of MUC5AC mRNA in lung tissue.

CONCLUSIONThese findings suggest that increased expression of a single-gene, mCLCA3, could simulate an asthma attack, and its mechanism may involve mCLCA3 overexpression up-regulating IL-13 expression.

Allergens ; Animals ; Asthma ; Chloride Channels ; Female ; Inflammation ; chemically induced ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; genetics ; metabolism ; Interleukin-5 ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mucin 5AC ; genetics ; metabolism ; Ovalbumin ; pharmacology

The present study aims to investigate the effect of Cbl-b, a member of E3 ubiquitin ligase family, on interleukin-1 (IL-1) pathway in synoviocytes. The protein expression levels of Cbl-b and IL-1-induced matrix metalloproteinase 13 (MMP-13) in synoviocytes were analyzed by Western blot. Collagen substrates were incubated with the conditioned medium collected from synoviocytes cultures and then subjected to SDS-PAGE for analysis of collagen degradation. The results showed that compared with wild-type cells, Cbl-b-deficient cells expressed more MMP-13 protein and had enhanced ability to degrade collagens under IL-1 stimulation. These data suggest that Cbl-b may negatively regulate IL-1-triggered degradation of collagen matrix in synoviocytes.
Collagen ; metabolism ; Epithelial Cells ; enzymology ; Humans ; Interleukin-1 ; metabolism ; Matrix Metalloproteinase 13 ; metabolism ; Proto-Oncogene Proteins c-cbl ; metabolism ; Signal Transduction

OBJECTIVETo investigate the dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of different dose of ulinastatin through animal experiment.

METHODS104 male SD rats were randomly assigned into the control group, ulinastatin control group, phosgene treatment groups and different dose of ulinastatin intervention groups, 8 rats each group. Treatment groups were dynamic constant exposure in phosgene, and immediately injected ulinastatin intraperitoneal, and then the experimental animal, the lung tissue biopsy, lung wet/dry ratio, RT-PCR detection, the plasma for detection of Bio-Plex 18 cytokines.

RESULTSCompared with the control group, plasma concentrations of IL-1α, IL-6, GM-CSF, TNF-α, INF-γ, MIP-3α, VEGF were increased significantly first (2 h), and gradually decreased with the passage of time , the difference was statistically significant (P < 0.05). Plasma concentrations of IL-4, IL-10 were decreased earlier (2h, 6 h) and increased later (24 h) (P < 0.05). The change of plasma concentration of IL-13 was not obvious earlier (2 h) and still rising later (24h), the difference was statistically significant (P < 0.05). After drug intervention, the levels of pro-inflammatory cytokines declined and the levels of anti-inflammatory cytokines raise by different degrees at different times in ulinastatin intervention groups, the difference was statistically significant. The degree of lung injury was improved than the phosgene treatment groups and better in high dose of ulinastatin intervention group. The expression of IL-10, IL-4, IL-13-mRNA of tissue increased in accordance with plasma results.

CONCLUSIONA group of cytokines are dynamicly changed in phosgene-induced lung injury by time. High dose of ulinastatin can improved phosgene-induced lung injury, regulate the synthesis and release of inflammatory cytokines and inhibit inflammatory react in a dose-dependent manner.

Animals ; Cytokines ; metabolism ; Glycoproteins ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; Interleukin-10 ; Interleukin-13 ; Interleukin-1alpha ; Interleukin-4 ; Interleukin-6 ; Lung ; Lung Injury ; chemically induced ; drug therapy ; Male ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha

OBJECTIVETo explore the effect of nuclear factor-kappa B (NF-κB) decoy oligodeoxynucleotides (ODN) on serum NF-κB, IL-10, IL-13 and pulmonary NF-κB protein expression in rabbits with severe lung contusion.

METHODSForty New Zealand rabbits were randomly divided into severe lung contusion group (group A, n=12), lung contusion with NF-κB scrambled decoy ODN group (group B, n=12), lung contusion with sense NF-κB decoy ODN group (group C, n=12), and normal control group (n=4). After establishment of the contusion injury model, the sense and scrambled NF-κB decoy ODN were infused into the rabbits via the jugular vein accordingly. Serum NF-κB, IL-10, and IL-13 and NF-κB protein expressions in the lung tissue were detected before and at 1, 2, 3, and 4 h after the contusion.

RESULTSTwo hours after sense NF-κB decoy ODN intervention, the expression of NF-κB began to decrease and reached the lowest level at 3 h; pulmonary IL-10 and IL-13 expressions decreased at 1 h after contusion, to the lowest level at 2 and 4 h, respectively. After sense NF-κB decoy ODN intervention, the expression of IL-10 and IL-13 increased and NF-κB protein expression decreased significantly in comparison with those in groups A and B (P<0.01).

CONCLUSIONSense NF-κB decoy ODN can significantly reduce the serum NF-κB expression, increase serum IL-10 and IL-13 levels and decrease pulmonary NF-κB protein expression in the early stages after severe lung contusion in rabbits.

Animals ; Contusions ; blood ; Female ; Interleukin-10 ; blood ; Interleukin-13 ; blood ; Lung ; drug effects ; metabolism ; Lung Injury ; blood ; NF-kappa B ; blood ; Oligodeoxyribonucleotides ; pharmacology ; Rabbits

OBJECTIVETo study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat.

METHODSThirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test.

RESULTSWound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05].

CONCLUSIONSMacrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.

Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, Myelomonocytic ; genetics ; metabolism ; Collagen ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukin-13 ; Interleukin-4 ; Macrophages ; Male ; Phenotype ; Rats ; Skin ; injuries ; Tumor Necrosis Factor-alpha ; blood ; Wound Healing ; genetics