Serum interleukin-33 combined with FEF75% z-score and FeNO improves the diagnostic accuracy of asthma in children

Abstract Objective To investigate the diagnostic efficacy of serum IL-33 single indicator and combined indicators for asthma in children. Methods 132 children were initially diagnosed with asthma during acute exacerbation and 100 healthy children were included. Serum IL-33 concentration differences were compared between asthmatic and normal children. Correlations between IL-33 with pulmonary function parameters, FeNO, peripheral blood EOS counts and serum total IgE were analyzed in asthmatic children. ROC curves were used to assess IL-33 diagnostic efficacy and its combined indicators. To prevent overfitting of the predictive model, the hold-out cross-validation method was used. Results (1) Serum IL-33 concentrations were significantly higher in children with asthma than in normal children (p < 0.001). (2) IL-33 concentration was negatively correlated with FVC z-score, FEV1 z-score and FEF75% z-score in asthmatic children (p < 0.05). (3) The area under the ROC curve of IL-33 was 0.821, which was higher than those of FeNO, FVC z-score, and FEV1 z-score. (4) Cross-validation of the combined indicators showed that IL-33 significantly improved asthma diagnostic efficacy. The combination of IL-33, FEF75% z-score, and FeNO showed the highest diagnostic efficacy, with the AUC, sensitivity, and specificity of the combined indicator being 0.954, 90.1%, and 89. 0%, respectively, and good extrapolation of the predictive model. Conclusion Serum IL-33 is higher in children with asthma and increases with the severity of pulmonary ventilation obstruction. A single indicator of serum IL-33 demonstrates moderate diagnostic accuracy, and its combination with FEF75% z-score and FeNO significantly improves the diagnostic accuracy in childhood asthma.

Evaluation of interleukin-33 & sST2 levels in type-2 diabetic mellitus patients with or without metabolic syndrome

Background & objectives: Diabetes mellitus (DM) is characterized by increase in blood glucose levels due to defective insulin secretion or insulin sensitivity. Interleukins (ILs) are known to play an important role in the pathogenesis of DM. The aim of this study was to investigate the serum concentration of IL-33 and its receptor soluble ST2 (sST2) in patients with diabetes and draw a correlation between their serum levels and different standard glycaemic indices of patients affected with type-2 diabetes with or without metabolic syndrome. Methods: Thirty type-2 diabetic individuals and 30 healthy controls were recruited for this study. Serum and plasma were separated by centrifugation of blood for quantitative measurement of IL-33, sST2 and other biochemical parameters. Results: It was observed that serum IL-33 levels were significantly less and sST2 levels were significantly high in type-2 diabetic individuals as compared to healthy controls. A significant correlation between the serum IL-33 concentration and fasting plasma glucose (FPG) and postprandial plasma glucose (PPG) levels were also found. Additionally, data also elucidated that serum levels of high-density lipoprotein, low-density lipoprotein or triglyceride in type-2 diabetics did not influence the serum levels of IL-33 and sST2, thereby excluding these factors as the major drivers of changes in serum IL-33 and sST2 concentration. Interpretation & conclusions: This study demonstrated alteration in serum levels of IL-33 and sST2 in type-2 diabetic individuals. Further mechanistic studies, focusing on the progression of type-2 diabetes could elucidate the involvement of IL-33 in the cellular acquisition of insulin resistance as observed in type-2 diabetics

Regulating the immune response to carbon tetrachloride-induced liver fibrosis in mice by blocking inducible co-stimulatory molecules and interleukin-33 / 中华肝脏病杂志

Objective: To investigate the effects of combined blockade of interleukin-33 (IL-33) and inducible co-stimulatory molecule (ICOS) on carbon tetrachloride-induced chronic liver fibrosis and imbalance of T helper lymphocyte subsets in mice. Methods: There were 40 BALB/c mice in each model and control group. Flow cytometry was used to determine the proportion of Th1/Th2/Th17 cells in the splenic lymphocyte suspension of mice, the expression levels of interferon γ, IL-4, and IL-17 in the splenic lymphocyte suspension of liver fibrosis mice after combined blockade of IL-33 and ICOS, and the pathological changes of liver histopathology in mice with liver fibrosis. Two independent sample t-test was used to compare data between groups. Results: Compared with the non-blocking group, the proportion of Th2 and Th17 cells in the IL-33/ICOS blocking group was significantly down-regulated (Th2: 65.96% ± 6.04% vs. 49.09% ± 7.03%; Th17: 19.17% ± 4.03% vs. 9.56% ± 2.03%), while the proportion of Th1 cells and Th1/Th2 ratio were up-regulated (Th1: 17.14% ± 3.02% vs. 31.93% ± 5.02%; Th1/Th2: 0.28 ± 0.06 vs. 0.62 ± 0.23), and the difference was statistically significant (t = 5.15, 6.03, 7.14, 4.28, respectively, with P < 0.05). After entering the chronic inflammation stage of liver fibrosis in mice (10 weeks), compared with the non-blocking group, the expression levels of IL-4 and IL-17 in the blockade group were significantly down-regulated [IL-4: (84.75 ± 14.35) pg/ ml vs. (77.88 ± 19.61) pg/ml; IL-17: (72.38 ± 15.13) pg/ml vs. (36.38 ± 8.65) pg/ml], while the expression of interferon γ was up-regulated [(37.25 ± 11.51) pg/ml vs. (77.88 ± 19.61) pg/ml], and the difference was statistically significant (t: IL-4: 4.71; IL-17: 5.84; interferon γ: 5.05, respectively, with P < 0.05). Liver histopathological results showed that hepatic necrosis, hepatic lobular structural disorder, and fibrous tissue hyperplasia were significantly lower in the blockade group than those in the non-blocking group at 13 weeks of liver fibrosis. Conclusion: Combined blockade of the ICOS signaling pathway and IL-33 can regulate Th2 and Th17 polarization, down-regulate the inflammatory response, and inhibit or prevent the occurrence and progression of fibrosis.

Correlation between Serum Interleukin-33, β2-Microglobulin Levels and DS Stage in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the correlation between serum interleukin-33 (IL-33), β2microglobulin (β2-MG) levels and Durie-Salmon (DS) stage in patients with multiple myeloma (MM).@*METHODS@#100 MM patients admitted to the First Affiliated Hospital of Fujian Medical University from March 2019 to January 2021 were selected and divided into stage I, stage II and stage III groups according to the DS staging system. A baseline data questionnaire of patients was designed, then the relevant baseline data and laboratory test results of patients were recorded. The levels of serum IL-33 and β2-MG of all patients were detected, and the correlation between serum IL-33, β2-MG levels and DS stage of MM patients was analyzed.@*RESULTS@#Among the 100 patients with MM, there were 32 cases in stage I, 39 cases in stage II and 29 cases in stage III. The levels of serum CRP and β2-MG of patients in stage III were significantly higher than those of patients in stage I and II, and the levels of serum CRP and β2-MG of patients in stage II were significantly higher than those of patients in stage I, the differences were statistically significant (P <0.05). The level of serum IL-33 of patients in stage III was significantly lower than that of patients in stage I and II, and the level of serum IL-33 of patients in stage II was significantly lower than that of patients in stage I, the differences were statistically significant (P <0.05). There was no statistical significant difference in other data between groups (P >0.05). Kendall's tau-b correlation analysis showed that the levels of serum CRP and β2-MG were positively correlated with DS stage in MM patients (r =0.534, 0.776), the level of serum IL-33 was negatively correlated with DS stage in MM patients (r =-0.759). Ordered logistic regression analysis and forest plot showed that the low level of serum IL-33 and the high level of β2-MG were the influencing factors of high DS stage in MM patients (P <0.05 ).@*CONCLUSION@#DS stage of MM patients is closely related to the levels of serum IL-33 and β2-MG, that is, the lower the serum IL-33 level and the higher the β2-MG level, and the higher the DS stage of MM patients.

Mechanism of epidural scar hyperplasia after laminectomy induced by blood siltation / 中华骨科杂志

Objective:To explore the mechanism of epidural scar tissue hyperplasia induced by erythrocyte rupture and release of interleukin-33 (IL-33) after laminectomy in mice.Methods:In the zoological experiment, the operation group (Laminectomy) and the sham operation group were set, and HE staining and Masson staining were performed to test for blood accumulation in the operation area after laminectomy in mice. Then 12 wild-type mice with 6-8 week old were selected and divided into 4 groups: the sham operation group, the operation group (normal saline control), the pure red blood cell intervention operation group, the whole blood intervention operation group. The normal saline (100 mg/kg) was injected into the postoperative area. The red blood cells or whole blood with the same volume were injected into the postoperative area in the pure red blood cell intervention group and the whole blood intervention group. The postoperative recovery of mice in each group was observed. The levels of fibronectin in the postoperative scar tissues of mice in four groups were detected by western blot technology, and the degree of postoperative epidural scar hyperplasia was directly observed by immunohistochemistry. In the cytological experiment, the wild-type mouse erythrocyte normal saline group, the control group of IL-33 knockout mouse erythrocyte normal saline, the wild-type mouse erythrocyte lysis group, and the IL-33 knockout mouse erythrocyte lysis group were set. The levels of IL-33 in the red blood cells of four groups were detected by western blot. Then, a blank wild-type mouse erythrocyte control group, a wild-type mouse relative to the control group (only secondary antibody added to test for non-specific binding), a wild-type mouse erythrocyte group and an IL-33 knockout mouse erythrocyte group (to test for antigen specificity of the primary antibody) were set. Immunofluorescence staining was performed on the erythrocytes of four groups and the level of IL-33 was detected by flow cytometry.Results:HE staining and Masson staining after laminectomy showed that there was blood stasis in the local incision area of mice in the operation group. The epidural scar hyperplasia in the incision area of mice after whole blood or red blood cells intervention was higher, especially in the whole blood intervention group. IL-33 expression was almost undetectable in the wild-type erythrocyte normal saline control group, the IL-33-knockout erythrocyte normal saline control group, and the IL-33-knockout erythrocyte lysis group, while significant IL-33 expression was detectable in the wild-type erythrocyte lysis group. Immunofluorescence staining showed that IL-33 was expressed in and on the erythrocyte membrane of wild-type mice, while non-specific expression of IL-33 or a very small amount of IL-33 was almost undetectable in the other three groups. The immunofluorescence intensities of IL-33 in the four groups were 0.62±0.41, 60.17±4.39, 16.78±7.43 and 0.61±0.03, respectively ( F=281.90, P<0.001). The expression of IL-33 in the erythrocyte group of wild-type mice was the highest ( P<0.05). According to the results of flow cytometry, except for the trace amount of IL-33 detected in the wild-type mouse erythrocyte group, the expression of IL-33 in the other three groups was basically 0. The ratios of fibronectin to β-actin in the modeling area of the four groups gradually increased, and the ratios were 0.79±0.09, 1.26±0.23, 1.79±0.05 and 2.29±0.58, respectively, and the differences were statistically significant ( F=12.86, P=0.002). Fibronectin in the operation area of the three operation groups (normal saline control group, red blood cell intervention group and whole blood intervention group) was significantly higher than that of the sham operation group. The immunohistochemical staining results of fibronectin in the modeling area of the four groups were the same as those in western blot experiment. The average optical density values of fibronectin in each group were 0.09±0.01, 0.18±0.01, 0.22±0.01 and 0.24±0.01, respectively, and the difference was statistically significant ( F= 210.7, P<0.001). Conclusion:There is indeed blood accumulation in the surgical area after laminectomy in mice, and it can aggravate the hyperplasia of epidural scar tissue. Erythrocyte is the main component in blood, and there is a large amount of IL-33 expression in the inner and outer membrane of erythrocyte membrane. The mechanism of promoting the proliferation of epidural scar tissue may be related to the release of IL-33 by erythrocyte lysis.

Effects of aerosol therapy with budesonide suspension combined with compound ipratropium bromide on partial pressure of carbon dioxide and tumor necrosis factor α in children with bronchiolitis / 中国基层医药

Objective:To investigate the effects of aerosol therapy with budesonide suspension combined with compound ipratropium bromide on partial pressure of carbon dioxide (PaCO 2) and tumor necrosis factor α (TNF-α) in children with bronchiolitis. Methods:A total of 124 children with bronchiolitis admitted to Gujiao Central Hospital from January 2019 to December 2021 were included in this study. These children were randomly divided into two groups using the coin-tossing method. The control group ( n = 62) was treated with routine symptomatic treatment, and the study group ( n = 62) was treated with aerosol therapy of budesonide suspension combined with compound ipratropium bromide based on routine symptomatic treatment. The time at which clinical symptoms disappear, clinical efficacy, inflammatory reaction, and blood gas index were determined in each group. Results:After treatment, the time at which asthma, cough, pulmonary rales, and fever in the study group were (2.28 ± 0.71) days, (3.30 ± 0.82) days, (5.25 ± 1.03) days, and (19.01 ± 2.65) hours, respectively, which were significantly shorter than (2.71 ± 0.89) days, (3.81 ± 0.98) days, (5.72 ± 1.37) days, and (20.76 ± 3.12) hours in the control group ( t = 2.97, 3.14, 2.15, 3.36, all P < 0.05). Total response rate and PaO 2 in the study group were 91.94% and (83.94 ± 4.02) mmHg, respectively, which were significantly higher than 77.42% and (81.25 ± 5.53) mmHg in the control group ( χ2 = 5.03, t = 3.09, both P < 0.05). Interleukin-18, interleukin-33, TNF-α, and PaCO 2 in the study group were (141.03 ± 34.69) ng/L, (143.87 ± 38.43) ng/L, (75.49 ± 18.43) ng/L, and (41.85 ± 3.31) mmHg, respectively, which were significantly lower than (158.64 ± 47.92) ng/L, (162.75 ± 50.32) ng/L, (83.22 ± 21.75) ng/L, and (43.58 ± 4.46) mmHg in the control group ( t = -2.34, -2.34, -3.23, -2.45, all P < 0.05). Conclusion:Aerosol therapy with budesonide suspension combined with compound ipratropium bromide based on routine symptomatic treatment is more effective on bronchiolitis than routine symptomatic treatment alone. The combined therapy can effectively decrease PaCO 2 and TNF-α levels.

Cerebrospinal fluid levels of sortilin-1, lipocalin-2, autotaxin, decorin and interleukin-33 in patients with idiopathic intracranial hypertension / Níveis de sortilina-1, lipocalina-2, autotaxina, decorina e interleucina-33 no líquido cefalorraquidiano em pacientes com hipertensão intracraniana idiopática

Abstract Background Idiopathic intracranial hypertension (IIH) is characterized by increased cerebrospinal fluid (CSF) pressure of unknown cause. It has been suggested that the inflammatory process plays a role in the pathophysiology of the disease. Sortilin-1, lipocalin-2, autotaxin, decorin, and interleukin-33 (IL-33) are among the factors involved in inflammatory processes. Objective To investigate the CSF levels of sortilin-1, lipocalin-2, autotaxin, decorin, and IL-33 in patients with IIH. Methods A total of 24 IIH patients and 21 healthy controls were included in the study. Demographic characteristics of the patients and of the control group as well as CSF pressures were evaluated. Sortilin-1, lipocalin-2, autotaxin, decorin and IL-33 levels in the CSF were measured. Results The CSF levels lipocalin-2, sortilin-1, autotaxin, IL-33 and CSF pressure were significantly higher in the patients group compared with the control group (p < 0.001). Decorin levels were reduced in patients (p < 0.05). There was no correlation between the autotaxin and IL-33 levels and age, gender, CSF pressure, and body mass index. The results of our study showed that inflammatory activation plays an important role in the development of the pathophysiology of IIH. In addition, the fact that the markers used in our study have never been studied in the etiopathogenesis of IIH is important in explaining the molecular mechanism of this disease. Conclusion Studies are needed to evaluate the role of these cytokines in the pathophysiology of the disease. It is necessary to evaluate the effects of these molecules on this process.

Resumo Antecedentes A hipertensão intracraniana idiopática (HII) é caracterizada pelo aumento da pressão do líquido cefalorraquidiano (LCR) de causa desconhecida. Tem sido sugerido que o processo inflamatório desempenha um papel na fisiopatologia da doença. Sortilina-1, lipocalina-2, autotaxina, decorina e interleucina-33 (IL-33) estão entre os fatores envolvidos nos processos inflamatórios. Objetivo Investigar os níveis de sortilina-1, lipocalina-2, autotaxina, decorina e IL-33 no LCR de pacientes com HII. Métodos Um total de 24 pacientes com HII e 21 controles saudáveis foram incluídos no estudo. Foram avaliadas as características demográficas dos pacientes e do grupo controle, bem como as pressões liquóricas. Os níveis de sortilina-1, lipocalina-2, autotaxina, decorina e IL-33 no LCR foram medidos. Resultados Os níveis no líquido cefalorraquidiano lipocalina-2, sortilina-1, autotaxina, IL-33 e pressão liquórica foram significativamente maiores no grupo de pacientes em comparação com o grupo controle (p < 0,001). Os níveis de decorina foram reduzidos nos pacientes (p < 0,05). Não houve correlação entre os níveis de autotaxina e IL-33 e idade, sexo, pressão liquórica e índice de massa corporal. Os resultados do nosso estudo mostraram que a ativação inflamatória desempenha um papel importante no desenvolvimento da fisiopatologia da HII. Além disso, o fato de os marcadores utilizados em nosso estudo nunca terem sido estudados na etiopatogenia da HII é importante para explicar o mecanismo molecular dessa doença. Conclusão Estudos são necessários para avaliar o papel dessas citocinas na fisiopatologia da doença. É necessário avaliar os efeitos dessas moléculas nesse processo

Evaluation of Serum Interleukin-33 and Soluble Suppression of Tumorigenicity 2 (sST2) Receptors in Patients with and without Periodontal Disease

Context: Interleukin?33 and its receptor soluble suppression of tumorigenicity 2 (sST2) play an important role in inflammation and its role in periodontal disease is yet unclear. The role of both IL?33 and sST2 together in periodontal disease as biomarkers has never been studied. Aim: To assess the levels of IL?33 and sST2 in serum samples of patients with periodontitis and healthy subjects. Methods: A total of 71 subjects (30 healthy subjects and 41 patients with periodontal disease) were included in the cross?sectional study. Community Periodontal Index (CPI) was used to assess periodontal health by utilizing a mouth mirror and a CPI probe. Venous blood was collected and serum was separated. Serum levels of IL?33 and sST2 were determined by the enzyme?linked immunosorbent assay (ELISA) assay. Statistical Analysis: Graph Pad Prism 5 was used for statistical analysis. Mann Whitney test was applied to compare the two groups. Results: The level of IL?33 was not found to be elevated among healthy subjects and sST2 was found elevated among patients with periodontal disease. The serum concentration of IL?33 was found at 472 ± 114 pg/ml and 282 ± 77 pg/ml among healthy subjects and patients with periodontal disease respectively. Significantly higher values of sST2 at 28 ± 2 ng/ml were found among periodontal patients as compared to healthy subjects with values of 18 ± 1 ng/ml. No significant differences were noted between mild to moderate and severe periodontitis for IL?33 and sST2 between the two groups. Conclusion: This study shows alteration in serum levels of IL?33 and sST2 in periodontitis patients. IL?33 and sST2 may be potential inflammatory markers of periodontitis. Further studies are required on a large sample size for better understanding. This pilot study is the first to assess the serum levels of both IL?33 and sST2 together among patients with and without periodontal disease.

Expression of interleukin-33 in hepatocellular carcinoma patients and its role in regulating CD8 / 临床肝胆病杂志

Objective To investigate the change in interleukin-33 (IL-33) in the peripheral blood of hepatocellular carcinoma (HCC) patients and the role and potential mechanism of IL-33 in regulating CD8 + T cell function in HCC patients. Methods A total of 44 HCC patients who attended Shaanxi Provincial People's Hospital from April 2019 to January 2020 and 20 healthy controls were enrolled. Peripheral blood was collected, and plasma and peripheral blood mononucleated cells (PBMCs) were isolated; ELISA was used to measure the plasma levels of IL-33 and its receptor ST2, and quantitative real-time PCR was used to measure the relative mRNA expression levels of IL-33 and ST2 in PBMCs. CD8 + T cells were purified and stimulated with recombinant IL-33; CCK-8 assay was used to assess cell proliferation, enzyme-linked immunospot assay was used to measure the secretion of perforin and granzyme B, and flow cytometry was used to measure the expression of PD-1, LAG-3, and CTLA-4; changes in cell proliferation, secretion of cytotoxic molecules, and immune checkpoint molecules after IL-33 stimulation were compared. CD8 + T cells were co-cultured with HepG2 cells; the expression of lactate dehydrogenase was measured to calculate the proportion of dead HepG2 cells induced by CD8 + T cells, and the change in the killing function of CD8 + T cells after IL-33 stimulation was compared. The t -test or the paired t -test was used for comparison of continuous data between two groups, and a Pearson correlation analysis was performed. Results Compared with the control group, the HCC group had significantly lower plasma level of IL-33 (269.80±63.08 pg/ml vs 339.50±64.43 pg/ml, t =4.072, P 0.05). The proportion of CD8 + T cells was not correlated with the plasma level of IL-33 or ST2 (both P > 0.05). Compared with the control group, the HCC group had significantly lower levels of perforin and granzyme B (both P 0.05), but it promoted the secretion of perforin and granzyme B ( P < 0.05). Compared with the control group, the HCC group had a significant reduction in the killing activity of CD8 + T cells ( P < 0.05), and stimulation with recombinant IL-33 enhanced the killing function of CD8 + T cells, which was mainly reflected in the increases in the proportion of dead HepG2 cells ( P < 0.05) and the secretion of IFNγ and TNFα ( P < 0.05). Conclusion There is a reduction in the plasma level of IL-33 in HCC patients. IL-33 can enhance the killing activity of CD8 + T cells by promoting the secretion of perforin and granzyme B, which provides a new target for the treatment of HCC.

Role of epithelium-derived cytokines interleukin-33, interleukin-25 and thymic stromal lymphopoietin in the pathogenesis of atopic dermatitis / 中华皮肤科杂志

Epidermal barrier defects and immune abnormalities are the main pathophysiological changes in the development of atopic dermatitis (AD) . Skin keratinocytes can release a variety of inflammatory factors and mediators under the treatment with various harmful factors. Three epithelium-derived cytokines interleukin (IL) -33, IL-25 and thymic stromal lymphopoietin are considered to be effective inducers of Th2 immune response in skin or mucosal barrier, which can activate immune cells, cause the secretion of Th2 cytokines, enhance the Th2 immune response, and participate in the occurrence and development of AD. This review focuses on the role of the above 3 epithelium-derived cytokines in the pathogenesis of AD.

Interleukin 33 inhibits lipopolysaccharide-induced high permeability of cardiac microvascular endothelial cells / 中华内科杂志

Objective:To investigate the effect of interleukin-33 (IL-33) on lipopolysaccharide (LPS)-induced permeability of rat cardiac microvascular endothelial cells (RCMECs).Methods:RCMECs were cultured in vitro to be divided into control group, LPS group, IL-33 group and LPS+IL-33 group. The effect of IL-33 on the proliferation of RCMECs was detected by cell counting reagent (CCK8). Fluorescein isothiocyanate (FITC)-dextran assay was used to evaluate the permeability of RCMECs. The expression of vascular endothelial calmodulin, ras homologous gene family (Rho) member A (RhoA) and phosphorylated Rho-associated coiled-coil-containing protein kinase (p-ROCK2) proteins were tested by western blot. High-throughput sequencing and gene ontology (GO) were performed for gene expression in LPS and LPS+IL-33 groups.Results:No significant effect of IL-33 at 10-50 ng/ml on the proliferation of RCMECs was observed ( P>0.05). Compared with the control group, the permeability of RCMECs (permeability coefficient ratio 1.404±0.029 vs. 1.000±0.200, P<0.05) was significantly increased in LPS group and the expression of vascular endothelial calmodulin (relative gray value 0.429 5±0.012 9 vs. 0.594 9±0.014 2, P<0.05) was down-regulated, while the permeability of monolayers (permeability coefficient ratio, 0.948±0.013, P<0.01) was decreased in LPS+IL-33 group and the expression of vascular endothelial calmodulin (relative grayscale value 0.549 1±0.012 0, P<0.005) was up-regulated compared with the LPS group. High-throughput sequencing data revealed that the differential genes downregulated in the LPS and LPS+IL-33 groups were associated with cytoskeleton and Rho signaling pathway. Compared with the control group, RhoA (relative gray value 0.211 4±0.009 9 vs. 0.135 0±0.007 6, P<0.000 1) and p-ROCK (relative gray value 0.656 3±0.013 2 vs. 0.503 6±0.036 2, P<0.000 1) protein expression was upregulated in the LPS group. When compared with LPS group, RhoA (relative gray value 0.157 7±0.010 7, P=0.000 2), p-ROCK (relative gray value 0.427 7±0.003 8, P<0.000 1) protein expression was decreased in LPS+IL-33 group. Conclusion:IL-33 may improve LPS-induced hyperpermeability of RCMECs by inhibiting RhoA and p-ROCK protein expression in Rho/Rho-associated coiled-coil-containing protein kinase signaling pathway.

Effect of Xiao Qinglongtang on IL-33/ST2 Signaling Pathway in Mice with Allergic Rhinitis / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effect of Xiao Qinglongtang （XQLT） on ovalbumin （OVA）-induced allergic rhinitis （AR） in mice and its effect on the interleukin-33 （IL-33）/suppression of tumorigenicity 2 （ST2） signaling pathway. MethodSeventy-two female BALB/c mice of SPF grade were randomly divided into a control group， a model group， a positive control group （loratadine， 2.05 mg·kg-1）， and low-， medium-， and high-dose （5.005，10.01，20.02 g·kg-1） XQLT groups. All mice except for those in the control group were sensitized by intraperitoneal injection of OVA solution， and the AR model was induced by intranasal drops of OVA solution. Thirty minutes before local intranasal drops， drugs were administered once， and mice in the control group and the model group received phosphate buffered saline （PBS） at 20 mL·kg-1 for 7 days. After the last intranasal drop of OVA solution， the times of sneezing and nasal rubbing of mice within 10 min was recorded. After drug administration for 7 days， blood samples were collected， and nasal bones of mice were decalcified for the preparation of pathological sections. The content of OVA-specific immunoglobulin E （OVA-sIgE）， interleukin-4 （IL-4）， interleukin-5 （IL-5）， and interleukin-13 （IL-13） was detected by enzyme-linked immunosorbent assay （ELISA） kits. Hematoxylin-eosin （HE） staining， periodic acid-Schiff （PAS） staining， and Giemsa staining were used to observe the pathological changes， goblet cell hyperplasia， and eosinophil infiltration of nasal mucosa， respectively. Western blot was used to detect the expression levels of IL-33， ST2， and IL-1 receptor accessory protein （IL-1RAP） in nasal mucosa. ResultCompared with the control group， the model group showed increased times of sneezing and nasal rubbing （P<0.01）， edema and thickening of nasal mucosa， goblet cell hyperplasia and eosinophil infiltration， increased serum levels of OVA-sIgE， IL-4， IL-5 and IL-13 （P<0.01）， and increased protein expression of IL-33， ST2， and IL-1RAP in nasal mucosa （P<0.05，P<0.01）. After drug administration， compared with the model group， the high-dose XQLT group showed reduced times of sneezing and nasal rubbing （P<0.01）， improved pathological conditions of nasal mucosa， reduced serum levels of OVA-sIgE， IL-4， IL-5， and IL-13 （P<0.01）， and declining protein expression of IL-33， ST2， and IL-1RAP in nasal mucosa （P<0.05，P<0.01）. ConclusionXQLT has a therapeutic effect on OVA-sensitized AR mice， and the mechanism may be related to the regulation of the IL-33/ST2 signaling pathway and Th2 inflammatory cytokine to reduce Th2 inflammatory response and alleviate nasal mucosal injury.

Urine IL-6, IL-8, IL-33 and MMP-9 levels in patients with IgA nephropathy and their predictive value for disease progression / 中国医师杂志

Objective:To investigate the urineinterleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-33 (IL-33) and matrix metalloproteinase-9 (MMP-9) in patients with IgA nephropathy (IgAN) and their predictive value for disease progression.Methods:110 IgAN patients admitted to Zhuozhou Hospital from January 2018 to October 2021 were selected and divided into IgAN progression group (39 cases) and IgAN non progression group (71 cases) according to the progress of IgAN patients. According to the Oxford Classification Standard System (MEST-C) of IgAN, they were divided into MEST-C≥3 group (42 cases) and MEST-C<3 group (68 cases). According to the estimated glomerular filtration rate (eGFR), they were divided into eGFR≥50 ml/(min·1.73 m 2) group (group A, 63 cases) and eGFR<50 ml/(min·1.73 m 2) group (group B, 47 cases). According to the amount of urinary protein, they were divided into urinary protein≥1.5 g/24 h group (66 cases) and urinary protein<1.5 g/24 h group (44 cases). Multivariate logistic regression was used to analyze the risk factors affecting the progression of IgAN. Receiver operating characteristic (ROC) curve was drawn to analyze the value of urinary IL-6, IL-8, IL-33 and MMP-9 levels in predicting the progression of IgAN. Results:The urinary IL-6, IL-8, IL-33 and MMP-9 levels in IgAN progression group were significantly higher than those in IgAN non progression group, while the serum albumin, eGFR and complement C3 in the IgAN progression group were lower than those in the IgAN non progression group (all P<0.05). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the MEST-C≥3 group were significantly higher than those in the MEST-C<3 group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the eGFR<50 ml/(min·1.73 m 2) group were significantly higher than those in the eGFR≥50 ml/(min·1.73 m 2) group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 in the urinary protein≥1.5 g/24 h group were significantly higher than those in the urinary protein<1.5 g/24 h group (all P<0.001). Multivariate logistic regression analysis showed that the course of disease, serum albumin, eGFR, urinary protein, IL-6, IL-8, IL-33 and MMP-9 were risk factors affecting the progression of IgAN (all P<0.005). The ROC curve showed that the area under the curve (AUC) of IL-6, IL-8, IL-33 and MMP-9 in predicting the progression of IgAN was 0.956 (95% CI: 0.891-0.998), and the sensitivity and specificity were 98.4% and 86.2%, respectively. Conclusions:The elevated levels of urinary IL-6, IL-8, IL-33 and MMP-9 are closely related to the progress of IgAN, and the combination of these four indicators has a good value in predicting the progress of IgAN.

Effect of IL-33 on pyroptosis of macrophages in mice with sepsis via NF-κB/p38 MAPK signaling pathway

ABSTRACT Purpose To demonstrate the effect of IL-33 on the macrophage pyroptosis in mice with sepsis through the NF-kB/p38 MAPK signal pathway. Methods In total, 24 C57BL/6 mice were divided into the sham operation group (sham) and the cecal ligation and puncture group (CLP). After CLP, 24 IL-33-/- mice were divided into the IL-33-/- group and the IL-33-/- intervention group. The latter group was intraperitoneally injected with IL-33. Mouse mortality was observed after CLP. Macrophage apoptosis in peritoneal lavage fluid was detected by flow cytometry. Serum inflammatory factor level was detected by ELISA. Apoptotic protein expression and NF-κB/p38 MAKP signaling pathway protein expression were detected by qRT-PCR and Western blot. Results Knocking out IL-33 significantly reduced the mortality of CLP mice, as well as the mRNA expression of IL-33 and the levels of serum inflammatory factors, including IL-33, IL-1β, and IL-18. It also reduced the rate of macrophage apoptosis and the expression of the apoptotic protein caspase-1 p10; increased the expression of IκBα; and reduced the protein expression of NF-κB and p38 MAPK. These effects were reversed after exogenous injection of IL-33. Conclusions IL-33 can increase the level of macrophage pyroptosis in mice with sepsis (by activating the NF-kB/p38MAPK signal pathway) and the mortality of these mice.

Dual role of interleukin-33 in tumors / 中南大学学报(医学版)

Interleukin-33 (IL-33) is a new member of the IL-1 cytokine family which plays roles in the nucleus as a nuclear factor and is released by damaged or necrotic cells to act as a cytokine. It can be released via damaged or necrotic cells and functions as a cytokine. The released IL-33 activates the downstream NF-κB and MAPKs signaling pathways through the isomers of the specific receptor ST2 and the interleukin-1 receptor accessory protein (IL-1RAcP), resulting in danger signals and the activated multiple immune responses. IL-33 is abnormally expressed in various tumors and involves in tumorigenesis, development, and metastasis. Moreover, IL-33 can play both pro-tumor and anti-tumor roles in the same type of tumor.

Function of IL-33 in Wound Age Estimation of Skin Wounds in Mice / 法医学杂志

Objective To explore the application value of interleukin-33 （IL-33） in wound age estimation in forensic practice by observing the sequential changes of IL-33 after skin wound. Methods Skin wound models were generated on the back of mice with a round file of 5 mm in diameter. Skin samples of the same size were taken from the same parts of mice in control group and injury group 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d and 10 d after skin wound. Hematoxylin-eosin （HE） staining method was applied to observe the morphological changes in the recovering process after skin wound. Western blotting, immunohistochemistry staining and double immunofluorescence staining methods were applied to detect the expression changes of IL-33 in the skin wound samples. Results The results of Western blotting showed that the expression of IL-33 protein decreased slightly at 3 h after skin wound, increased gradually at 6 h after skin wound, and reached the peak value at 3 d, then decreased gradually. Immunohistochemistry staining results showed that faint positive expression of IL-33 was observed in epidermis, hair follicles, sebaceous glands and dermal resident cells of the control group skin. The positive cell rate of IL-33 increased at 3 h after skin wound and reached the peak value at 3 d, then decreased gradually. The results of double immunofluorescence staining showed that the majority of IL-33 positive cells from 1 d to 3 d after wound were macrophages, while the majority of IL-33 positive cells from 5 d to 7 d after wound were myofibroblasts. In addition, the results of HE staining showed that the wound healing process of the skin wound model was consistent with the pathological development law of inflammation. Conclusion IL-33 could become a reference index for wound age estimation of skin wound in forensic practice.

Serum levels of interleukin-18, interleukin-33 and fractional exhaled nitric oxide in children with acute respiratory syncytial virus bronchiolitis / 中国医师进修杂志

Objective To observe the changes of interleukin-18 (IL-18),interleukin-33 (IL-33) and fractional exhaled nitric oxide (FeNO) in children with acute respiratory syncytial virus (RSV) bronchiolitis,and explore the potential mechanism of the transformation from acute RSV bronchiolitis to recurrent wheezing.Methods Fifty-three children with RSV bronchiolitis (RSV bronchiolitis group),32 children with repeated wheeze (repeated wheeze group) and 30 children receiving regular physical examination (healthy control group) from January 2016 to January 2017 in Cangzhou People's Hospital of Hebei Province were selected.The serum IL-18 and IL-33 at the time of inclusion and 2,3 months after inclusion were detected by enzyme linked immunosorbent assay,the FeNO at the same time was detected by multiple breathing technique,and the indexes were compared.The correlation between FeNO and IL-33,IL-18 was analyzed by Spearman method.Results The IL-18 at the time of inclusion and 2,3 months after inclusion in RSV bronchiolitis group and repeated asthmatic group were significantly higher than those in healthy control group:(10.89 ± 1.54) and (14.86 ± 5.54) ng/L vs.(7.26 ± 3.25) ng/L,(13.74 ± 4.16) and (15.45 ± 5.75) ng/L vs.(7.28 ± 3.56) ng/L,(11.38 ± 6.21) and (14.86 ± 5.28) ng/L vs.(7.18 ± 3.41) ng/L,those in repeated asthmatic group were significantly higher than those in RSV bronchiolitis group,and there were statistical differences (P＜0.05).The IL-33 levels at the time of inclusion and 2,3 months after inclusion in RSV bronchiolitis group and repeated asthmatic group were significantly higher than those in healthy control group:(17.68 ± 5.25) and (13.14 ± 5.01) ng/L vs.(3.69 ± 1.61) ng/L,(15.68 ± 4.16) and (15.11 ± 5.24) ng/L vs.(3.28 ± 1.56) ng/L,(13.87 ± 6.21) and (14.11 ± 5.14) ng/L vs.(3.18 ± 1.41) ng/L,IL-33 levels at the time of inclusion in RSV bronchiolitis group were significantly higher than those in repeated asthmatic group,and there were statistical differences (P＜0.05).The FeNO levels at the time of inclusion and 2,3 months after inclusion in repeated asthmatic group were significantly higher than those in RSV bronchiolitis group and healthy control group:(13.14 ± 4.47) ppb vs.(1.89 ± 1.54) and (7.26 ± 4.25) ppb,(14.75 ± 5.15) ppb vs.(7.74 ± 4.16) and (7.28 ± 4.12) ppb,(13.68 ± 5.62) ppb vs.(11.38 ± 6.21) and (7.18 ± 3.41) ppb;compared with that in healthy control group,FeNO at the time of inclusion in RSV bronchiolitis group was significantly decreased,and at 3 months was significantly increased,and there were statistical differences (P＜0.05).Correlation analysis result showed that FeNO at 2 and 3 months after inclusion in RSV bronchiolitis group had positive correlation with IL-18 level at the time of inclusion and 2,3 months after inclusion (P＜0.05),and negative correlation with IL-33 (P＜0.05);FeNO at 2 and 3 months after inclusion in repeated asthmatic group showed positive correlation with IL-18 at the same time (P＜0.05),and was negatively correlated with IL-33 level (P＜0.05);there was no correlation between FeNO and IL-18,IL-33 in healthy control group (P＞0.05).Conclusions IL-18 and IL-33 may be involved in the pathogenesis of acute RSV bronchiolitis and recurrent wheezing,and the concentration of IL-18 and IL-33 is correlated with the level of FeNO.Its potential mechanism needs further study.

Correlation of serum interleukin-33 and osteoprotegerin levels with coronary artery disease and risk degree / 中国医师进修杂志

Objective To investigate the relationship between serum interleukin (IL)-33,osteoprotegerin and the severity of coronary artery disease in patients with coronary heart disease,and analyze the correlation between IL-33 and osteoprotegerin.Methods Two hundred and nineteen patients with coronary heart disease from April 2018 to June 2019 in Dalian Friendship Hospital were selected.The percutaneous coronary angiography was performed in all patients.Among them,acute myocardial infarction (AMI) was in 59 cases (AMI group),unstable angina pectoris (UAP) in 55 cases (UAP group),stable angina pectoris (SAP) in 55 cases (SAP group),and basically normal result was in 50 cases (control group).Serum IL-33 and osteoprotegerin levels were measured by enzyme-linked immunosorbent assay (ELISA),and the severity degree of coronary artery disease was assessed by Gensini scoring system.Results The IL-33 in SAP group,UAP group and AMI group was significantly higher than that in control group:(330.42 ± 82.56),(363.54 ± 61.36) and (448.62 ± 71.60) ng/L vs.(275.96 ± 74.34) ng/L,IL-33 in AMI group was significantly higher than that in SAP group and UAP group,and there were statistical differences (P ＜ 0.05).The osteoprotegerin in UAP group and AMI groupwas significantly higher than that in control group:(481.35 ± 101.68) and (558.29 ± 136.45) ng/L vs.(392.21 ± 109.57) ng/L,osteoprotegerin in AMI group was significantly higher than that in SAP group and UAP group:(558.29 ± 136.45) ng/L vs.(436.13 ± 121.84) and (481.35 ± 101.68) ng/L,and there were statistical differences (P＜ 0.05).The Gensini score in SAP group,UAP group and AMI group was significantly higher than that in control group:(23.31 ± 7.88),(44.37 ± 14.15) and (90.20 ± 42.01) scores vs.(7.17 ± 4.85) scores,Gensini score in UAP group was significantly higher than that in SAP group,Gensini score in AMI group was significantly higher than that in SAP group and UAP group,and there were statistical differences (P ＜ 0.05).Pearson correlation analysis result showed that serum IL-33 and osteoprotegerin levels were positively correlated with Gensini score (r =0.588 and 0.420,P ＜ 0.01),and serum IL-33 level was positively correlated with osteoprotegerin level (r =0.718,P ＜ 0.01).The receiver operating characteristic curve analysis result showed that the area under curve of IL-33 for the diagnosis of AMI was 0.887 (95％ CI 0.839 to 0.935,P ＜ 0.01),and the area under curve of osteoprotegerin for diagnosis of AMI was 0.754 (95％ CI 0.682 to 0.825,P＜0.01).Conclusions Serum IL-33 and osteoprotegerin levels are elevated in patients with coronary heart disease,and their elevated levels are positively correlated with the severity and instability of coronary artery disease and have a certain diagnostic value for AMI.

Correlation of serum interleukin-33 and osteoprotegerin levels with coronary artery disease and risk degree / 中国医师进修杂志

Objective@#To investigate the relationship between serum interleukin (IL)-33, osteoprotegerin and the severity of coronary artery disease in patients with coronary heart disease, and analyze the correlation between IL-33 and osteoprotegerin.@*Methods@#Two hundred and nineteen patients with coronary heart disease from April 2018 to June 2019 in Dalian Friendship Hospital were selected. The percutaneous coronary angiography was performed in all patients. Among them, acute myocardial infarction (AMI) was in 59 cases (AMI group), unstable angina pectoris (UAP) in 55 cases (UAP group), stable angina pectoris (SAP) in 55 cases (SAP group), and basically normal result was in 50 cases (control group). Serum IL-33 and osteoprotegerin levels were measured by enzyme-linked immunosorbent assay (ELISA), and the severity degree of coronary artery disease was assessed by Gensini scoring system.@*Results@#The IL-33 in SAP group, UAP group and AMI group was significantly higher than that in control group: (330.42 ± 82.56), (363.54 ± 61.36) and (448.62 ± 71.60) ng/L vs. (275.96 ± 74.34) ng/L, IL-33 in AMI group was significantly higher than that in SAP group and UAP group, and there were statistical differences (P<0.05). The osteoprotegerin in UAP group and AMI group was significantly higher than that in control group: (481.35 ± 101.68) and (558.29 ± 136.45) ng/L vs. (392.21 ± 109.57) ng/L, osteoprotegerin in AMI group was significantly higher than that in SAP group and UAP group: (558.29 ± 136.45) ng/L vs. (436.13 ± 121.84) and (481.35 ± 101.68) ng/L, and there were statistical differences (P<0.05). The Gensini score in SAP group, UAP group and AMI group was significantly higher than that in control group: (23.31 ± 7.88), (44.37 ± 14.15) and (90.20 ± 42.01) scores vs. (7.17 ± 4.85) scores, Gensini score in UAP group was significantly higher than that in SAP group, Gensini score in AMI group was significantly higher than that in SAP group and UAP group, and there were statistical differences (P<0.05). Pearson correlation analysis result showed that serum IL-33 and osteoprotegerin levels were positively correlated with Gensini score (r = 0.588 and 0.420, P<0.01), and serum IL-33 level was positively correlated with osteoprotegerin level (r = 0.718, P<0.01). The receiver operating characteristic curve analysis result showed that the area under curve of IL-33 for the diagnosis of AMI was 0.887 (95% CI 0.839 to 0.935, P<0.01), and the area under curve of osteoprotegerin for diagnosis of AMI was 0.754 (95% CI 0.682 to 0.825, P<0.01).@*Conclusions@#Serum IL-33 and osteoprotegerin levels are elevated in patients with coronary heart disease, and their elevated levels are positively correlated with the severity and instability of coronary artery disease and have a certain diagnostic value for AMI.

Association Between Epithelial Cytokines and Clinical Phenotypes of Elderly Asthma

PURPOSE: Asthma in the elderly has different clinical features including more severe phenotypes with higher comorbidities. Epithelial cells are known to initiate innate/adaptive immune responses in asthmatic airways. We investigated clinical features and epithelial derived cytokine levels in elderly asthmatics compared to non-elderly asthmatics in a cross-sectional cohort of adult asthmatics in order to further understand its pathogenic mechanisms. METHODS: A total of 1,452 adult asthmatics were enrolled from a single tertiary hospital and were classified into 2 groups: 234 elderly (≥ 60 years at initial diagnosis) and 1,218 non-elderly (< 60 years at initial diagnosis) asthmatics. Asthma-related clinical parameters were compared between the 2 groups. Serum levels of epithelial cell-derived cytokines including interleukin (IL)-31, IL-33, IL-8, eotaxin-2, transforming growth factor beta 1 (TGF-β1) and periostin were measured by enzyme-linked immunosorbent assay. RESULTS: Significantly higher prevalence rates of late-onset asthma (onset age ≥ 40 years) and severe asthma, as well as the lower rate of atopy, blood/sputum eosinophil counts, total immunoglobulin E and eosinophil cationic protein levels were noted in elderly asthmatics compared to non-elderly asthmatics (P < 0.05, respectively). The forced expiratory volume in 1 second (FEV1, % predicted) level tended to be lower in elderly asthmatics (P = 0.07). In addition, serum IL-33 and IL-31 levels were significantly lower in elderly asthmatics, while no differences were found in the serum level of IL-8, eotaxin-2, TGF-β1 or periostin. Among elderly asthmatics, subjects with severe asthma had lower FEV1 (% predicted) value, but showed significantly higher serum levels of eotaxin-2 and TGF-β1, than those with non-severe asthma (P < 0.05 for each). CONCLUSIONS: These findings suggest that age-related changes of epithelial cell-derived cytokines may affect clinical phenotypes and severity of elderly asthma: decreased levels of IL-33 and IL-31 may contribute to less Th2 phenotype, while increased levels of eotaxin-2 and TGF-β1 may contribute to severity.