The RAW264.7/THP1 cells have been extensively employed as macrophage models in inflammation research. However, comprehensive comparisons of their expression profiles remain largely unexplored. In this study, we conducted a systematic bioinformatics analysis to characterize the whole-cell proteome, phosphoproteome, acetylome, and ubiquitinome profiles of lipopolysaccharide-stimulated RAW264.7/THP1 cells. Through comparative temporal analysis of differentially regulated proteins (classified as rapid, persistent, or slow dysregulation patterns), we identified IGF2, COPZ1, and GTF2B, exhibiting persistent downregulation in both cell models. Additionally, we observed that nearly all dysregulated proteins within each modification type exhibited significant enrichment in the other two modification types, forming a highly interconnected protein-protein interaction network. Notably, VIMENTIN demonstrated significant downregulation across three modification types (phosphorylation at Ser56/214, acetylation at Lys235, and ubiquitination at Lys139), with coordinated downregulation observed in both cell lines. It was also shown that VIMENTIN mutation at Ser56, Lys235, and Lys139 dramatically reduces the expression of CDK1, which may help the inflammation response by averting mitotic catastrophe and changing the macrophage cell cycle. These findings provide valuable molecular insights into species-specific macrophage responses and establish an important reference data set for discovering novel inflammatory proteins and investigating macrophage-associated diseases using these experimental models. Notably, our work delineates vimentin-centered regulatory networks that may offer novel diagnostic biomarkers and therapeutic targets for inflammatory diseases.

Background The clinical efficacy of tumor necrosis factor inhibitors (TNFi) in axial spondyloarthritis (axSpA) is limited by high non-response rates, necessitating accurate pre-treatment stratification. This study aimed to develop and validate MRI-based interpretable deep learning radiomics (DLR) models to predict treatment response to TNFi in patients with axSpA. Methods In this prospective study, patients diagnosed with axSpA who underwent sacroiliac joint MRI before TNFi initiation were enrolled. Patients were allocated to training and test sets at an approximate 4:1 ratio. The endpoints were major (ASAS40) and moderate (ASAS20) improvement, as defined by the Assessment of SpondyloArthritis International Society criteria. Deep learning and radiomics features were extracted from MRI. A four-step selection process, including reproducibility analysis, univariable filtering, redundancy reduction, and LASSO regression, was used to distill robust predictors. These were then integrated with clinical data to build DLR-clinical (DLRC) models using L2-regularized logistic regression. Results A total of 183 patients (mean age, 26.5 ± 9.0 years; 121 [66.1%] males) were analyzed. The DLRC models were validated on the independent test set, with high area under the receiver operating characteristic curve (AUC) values for predicting ASAS40 (0.876, 95%CI: 0.738–0.973) and ASAS20 (0.886, 95%CI: 0.753–0.978) responses. Calibration curves indicated good model agreement, and decision curve analysis (DCA) confirmed their clinical utility. Compared to the clinical-only model, the DLRC models showed significant integrated discrimination improvement (IDI) ASAS40 and ASAS20 (0.234 and 0.283, all p < 0.05) prediction, respectively. Conclusion An MRI-based interpretable DLR approach accurately predicts TNFi treatment response in axSpA patients, offering a non-invasive tool to guide personalized therapy.

BACKGROUND:The clinical efficacy of tumor necrosis factor inhibitors (TNFi) in axial spondyloarthritis (axSpA) is limited by high non-response rates, necessitating accurate pre-treatment stratification. This study aimed to develop and validate MRI-based interpretable deep learning radiomics (DLR) models to predict treatment response to TNFi in patients with axSpA. METHODS:In this prospective study, patients diagnosed with axSpA who underwent sacroiliac joint MRI before TNFi initiation were enrolled. Patients were allocated to training and test sets at an approximate 4:1 ratio. The endpoints were major (ASAS40) and moderate (ASAS20) improvement, as defined by the Assessment of SpondyloArthritis International Society criteria. Deep learning and radiomics features were extracted from MRI. A four-step selection process, including reproducibility analysis, univariable filtering, redundancy reduction, and LASSO regression, was used to distill robust predictors. These were then integrated with clinical data to build DLR-clinical (DLRC) models using L2-regularized logistic regression. RESULTS:A total of 183 patients (mean age, 26.5 ± 9.0 years; 121 [66.1%] males) were analyzed. The DLRC models were validated on the independent test set, with high area under the receiver operating characteristic curve (AUC) values for predicting ASAS40 (0.876, 95%CI: 0.738-0.973) and ASAS20 (0.886, 95%CI: 0.753-0.978) responses. Calibration curves indicated good model agreement, and decision curve analysis (DCA) confirmed their clinical utility. Compared to the clinical-only model, the DLRC models showed significant integrated discrimination improvement (IDI) for ASAS40 and ASAS20 (0.234 and 0.283, all p < 0.05) prediction, respectively. CONCLUSION:MRI-based interpretable DLR models accurately predicted TNFi treatment response in axSpA patients, offering a non-invasive tool to guide personalized therapy.

To develop a machine learning (ML)-based model using MRI and clinical risk factors to enhance diagnostic accuracy for axial spondyloarthritis (axSpA). We retrospectively analyzed datasets from four centers (A–D), focusing on patients with chronic low back pain. A subset from center A was used for prospective validation. A deep learning (DL) model based on ResNet50 was constructed using sacroiliac joint MRI. Clinical variables were integrated with DL scores in ML algorithms to distinguish axSpA from non-axSpA patients. Model performance was assessed by the area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and accuracy. The study included 1294 patients (median age 31 years [interquartile range 24–42]; 35.5

BACKGROUND:Psoriasis is a common chronic inflammatory disease with an unclear aetiology. Keratinocytes in psoriasis are susceptible to exogenous triggers that induce inflammatory cell death. OBJECTIVES:To investigate whether gasdermin E (GSDME)-mediated pyroptosis in keratinocytes contributes to the pathogenesis of psoriasis. METHODS:Skin samples from patients with psoriasis and from healthy controls were collected to evaluate the expression of GSDME, cleaved caspase-3 and inflammatory factors. We then analysed the data series GSE41662 to further compare the expression of GSDME between lesional and nonlesional skin samples in those with psoriasis. In vivo, a caspase-3 inhibitor and GSDME-deficient mice (Gsdme-/-) were used to block caspase-3/GSDME activation in an imiquimod-induced psoriasis model. Skin inflammation, disease severity and pyroptosis-related proteins were analysed. In vitro, tumour necrosis factor (TNF)-α-induced caspase-3/GSDME-mediated pyroptosis in the HACAT cell line was explored. RESULTS:Our analysis of the GSE41662 data series found that GSDME was upregulated in psoriasis lesions vs. normal skin. High levels of inflammatory cytokines such as interleukin (IL)-1β, IL-6 and TNF-α were also found in psoriasis lesions. In mice in the Gsdme-/- and caspase-3 inhibitor groups, the severity of skin inflammation was attenuated and GSDME and cleaved caspase-3 levels decreased after imiquimod treatment. Similarly, IL-1β, IL-6 and TNF-α expression was decreased in the Gsdme-/- and caspase-3 inhibitor groups. In vitro, TNF-α induced HACAT cell pyroptosis through caspase-3/GSDME pathway activation, which was suppressed by blocking caspase-3 or silencing Gsdme. CONCLUSIONS:Our study provides a novel explanation of TNF-α/caspase-3/GSDME-mediated keratinocyte pyroptosis in the initiation and -acceleration of skin inflammation and the progression of psoriasis.

Background: Axial spondyloarthritis (axSpA) is frequently diagnosed late, particularly in human leukocyte antigen (HLA)-B27-negative patients, resulting in a missed opportunity for optimal treatment. This study aimed to develop an artificial intelligence (AI) tool, termed NegSpA-AI, using sacroiliac joint (SIJ) magnetic resonance imaging (MRI) and clinical SpA features to improve the diagnosis of axSpA in HLA-B27-negative patients. Methods: We retrospectively included 454 HLA-B27-negative patients with rheumatologist-diagnosed axSpA or other diseases (non-axSpA) from the Third Affiliated Hospital of Southern Medical University and Nanhai Hospital between January 2010 and August 2021. They were divided into a training set (n=328) for 5-fold cross-validation, an internal test set (n=72), and an independent external test set (n=54). To construct a prospective test set, we further enrolled 87 patients between September 2021 and August 2023 from the Third Affiliated Hospital of Southern Medical University. MRI techniques employed included T1-weighted (T1W), T2-weighted (T2W), and fat-suppressed (FS) sequences. We developed NegSpA-AI using a deep learning (DL) network to differentiate between axSpA and non-axSpA at admission. Furthermore, we conducted a reader study involving 4 radiologists and 2 rheumatologists to evaluate and compare the performance of independent and AI-assisted clinicians. Results: NegSpA-AI demonstrated superior performance compared to the independent junior rheumatologist (<= 5 years of experience), achieving areas under the curve (AUCs) of 0.878 [95% confidence interval (CI): 0.786-0.971], 0.870 (95% CI: 0.771-0.970), and 0.815 (95% CI: 0.714-0.915) on the internal, external, and prospective test sets, respectively. The assistance of NegSpA-AI promoted discriminating accuracy, sensitivity, and specificity of independent junior radiologists by 7.4-11.5%, 1.0-13.3%, and 7.4-20.6% across the 3 test sets (all P<0.05). On the prospective test set, AI assistance also improved the diagnostic accuracy, sensitivity, and specificity of independent junior rheumatologists by 7.7%, 7.7%, and 6.9%, respectively (all P<0.01). Conclusions: The proposed NegSpA-AI effectively improves radiologists' interpretations of SIJ MRI and rheumatologists' diagnoses of HLA-B27-negative axSpA.

Background: Ferroptosis is a type of regulated necrosis and glutathione peroxidase 4 (GPX4) has been recognized as a key enzyme that protects against ferroptosis. However, the significance of GPX4 in polymorphonuclear neutrophils (PMNs) of systemic lupus erythematosus (SLE) has not been explored. So we examined GPX4 mRNA in PMNs and analyzed its association with serological and clinical features. Methods: A single center research from the Department of Rheumatology and Immunology of the Third Affiliated Hospital, Southern Medical University was conducted between December 2020 and September 2022. Real-time transcription-polymerase chain reaction analysis was used to determine the expression of GPX4 mRNA in PMNs from patients suffering from different rheumatoid immune diseases and healthy controls. The associations of GPX4 RNA levels in SLE patients with serological and clinical indicators were assessed by Spearman's correlation analysis. Receiver operating characteristic curve analysis was performed to evaluate the diagnostic value of GXP4 mRNA for SLE. Results: GPX4 mRNA levels were lower in SLE patients than those in healthy individuals, patients with rheumatoid arthritis and axial spondyloarthritis. GPX4 mRNA levels were negatively correlated with the number of positive autoantibodies (r = -0.3072, p = 0.0300), anti-dsDNA antibody (r = -0.3654, p = 0.0336), antinucleosome antibody (r = -0.4052, p = 0.0263), erythrocyte sedimentation rate (r = -0.3773, p = 0.0069), C-reactive protein (r = -0.4037, p = 0.0036), and SLE disease activity index (r = -0.3072, p = 0.0300). GPX4 mRNA levels were downregulated in patients with alopecia compared with patients without alopecia (p < 0.05). The diagnostic capacity of GPX4 mRNA achieved high diagnostic accuracy (area under the curve = 0.848) with sensitivity (78.00%) and specificity (80.95%). Conclusion: Downregulated GPX4 mRNA in PMNs of SLE patients is negatively associated with the production of antinuclear antibodies, disease activity, and lupus alopecia, suggesting an important role of ferroptosis in SLE, high diagnostic value of GPX4 mRNA in PMNs, and potential therapies targeting GPX4 for SLE patients, especially lupus alopecia.

During the inflammatory storm of sepsis, a significant quantity of neutrophil extracellular traps (NETs) are generated, which act as a double-edged sword and not only impede the invasion of foreign microorganisms but also exacerbate organ damage. This study provides evidence that NETs can cause damage to alveolar epithelial cells in vitro. The sepsis model developed in this study showed a significant increase in NETs in the bronchoalveolar lavage fluid (BALF). The development of NETs has been shown to increase the lung inflammatory response and aggravate injury to alveolar epithelial cells. Bay-117082, a well-known NF-κB suppressor, is used to modulate inflammation. This analysis revealed that Bay-117082 efficiently reduced total protein concentration, myeloperoxidase activity, and inflammatory cytokines in BALF. Moreover, Bay-117082 inhibited the formation of NETs, which in turn prevented the activation of the pore-forming protein gasdermin D (GSDMD). In summary, these results indicated that excessive NET production during sepsis exacerbated the onset and progression of acute lung injury (ALI). Therefore, Bay-117082 could serve as a novel therapeutic approach for ameliorating sepsis-associated ALI.

Diffuse alveolar haemorrhage (DAH) is a rapidly developing condition owing to a lack of effective treatment and resulting in a high mortality rate in systemic lupus erythematosus (SLE). Neutrophil extracellular traps (NETs) contain numerous antigens and proinflammatory substances that directly damage the vascular endothelium and aggravate vascular inflammation, which is considered an important pathogenic factor of DAH in SLE. Therefore, blocking the release of NETs from neutrophils is an important target for the treatment of DAH in SLE. In this study, we investigated whether the inhibition of neutrophils releasing NETs could relieve DAH in SLE. Necrostatin-1 (Nec-1), a small molecule, has been reported to inhibit the release of NETs by neutrophils. In vitro experiments revealed that Nec-1 inhibited alveolar epithelial cell damage by preventing the release of NETs. Furthermore, vivo studies showed that Nec-1 alleviated lupus pulmonary haemorrhage in mice by reducing lung pathology severity, body weight, and serum inflammatory cytokine levels. Mechanistically, Nec-1 prevented NET release by inhibiting neutrophil elastase (NE) activation and N-Gasdermin D (N-GSDMD) expression. Additionally, immunohistochemistry and immunofluorescence findings showed that Nec-1 decreased NE expression in the lung tissues of mice with lupus pulmonary haemorrhage. Thus, NETs released by neutrophils contributed to the pathogenesis of DAH in SLE, and Nec-1 showed protective effects by the inhibition of NET production via the reduction of NE activation and N-GSDMD expression.

Background To investigate the effect and mechanism of Celastrol on the formation of neutrophil extracellular traps (NETs), and to provide a theoretical basis for the clinical application of Tripterygium wilfordii. Methods First, we isolated neutrophils from the peripheral blood of healthy volunteers, and then observed the effect of Celastrol on Phorbol Myristate Acetate (PMA)-induced neutrophil release of NETs. The level of NETs was detected by using the membrane-impermeable nucleic acid dye, SytoxGreens. In addition, the levels of reactive oxygen species (ROS) were also examined to determine whether Celastrol affects ROS production during PMA-induced NETs. Results Celastrol produced significant cytotoxicity at a concentration of 5 µM (213.2±75.07), and the effect of stimulant PMA (25 nM) treatment was not statistically different (197.3±25.15) (P=0.9167). Celastrol (1.25, 0.625, and 0.3125 µM) did not exhibit cytotoxicity when treating neutrophils. Compared with the PMA (25 nM) + Celastrol (1.25, 0.625, and 0.3125 µM) group and the PMA (25 nM) monotherapy group, SytoxGreen showed a statistically significant reduction in fluorescence at 528 µM under 485 µM light excitation. Also, under the co-localization marker of Hochest and SytoxGreen double staining, we observed that the release of NETs in the PMA-treated group was higher than that in the control group. The PMA-induced neutrophil release of NETs was markedly reduced compared to the PMA-treated group. The NET release was substantially decreased under double staining with the Hochest and SytoxGreen co-localization markers. The fluorescence intensity of the Celastrol plus PMA group was significantly lower than that of the PMA treatment group alone, indicating a decrease in the level of intracellular ROS. Interestingly, the level of ROS in the treatment group who received Celastrol alone was lower than that in the control group, indicating that Tripterygium wilfordii could inhibit the spontaneous production of ROS by neutrophils in the absence of stimulation. Conclusions The molecular mechanism of Celastrol involves inhibition of PMA-stimulated neutrophil NETs formation in vitro, which is possibly related to the reduction of ROS levels. This indicates that Celastrol, the main component in Tripterygium wilfordii, can inhibit the formation of NETs, which provides a theoretical basis for the study of NETs-related diseases.

Neutrophils release neutrophil extracellular traps (NETs) to trap pathogenic microorganisms. NETs are involved in the inflammatory response and bacterial killing and clearance. However, their excessive activation can lead to an inflammatory storm in the body, which may damage tissues and cause organ dysfunction. Organ dysfunction is the main pathophysiological cause of sepsis and also a cause of the high mortality rate in sepsis. Acute lung injury caused by sepsis accounts for the highest proportion of organ damage in sepsis. NET formation can lead to the development of sepsis because by promoting the release of interleukin-1 beta, interleukin-8, and tumor necrosis factor-alpha, thereby accelerating acute lung injury. In this review, we describe the critical role of NETs in sepsis-associated acute lung injury and review the current knowledge and novel therapeutic approaches.

Background:This study sought to assess the relationship between suppressor of cytokine signaling 3 (SOCS3) expression, SOCS3 promoter methylation status, and platinum-based chemotherapy responses in advanced non-small cell lung cancer (NSCLC) patients.Methods:A total of 400 advanced NSCLC patients with inoperable disease were enrolled in this study. All the patients underwent platinum-based chemotherapy treatment, and the clinical and prognostic outcomes of these patients were analyzed. The SOCS3 protein expression and SOCS3 promoter methylation status of the tumor tissues in these patients were also tested by immunohistochemistry and polymerase chain reaction (PCR), respectively. In addition, we knocked down SOCS3 expression via small-interfering RNA (siRNA) in the lung cancer cell lines and conducted in vitro analyses to examine cell viability and apoptosis.Results:Patients with higher expression levels of SOCS3 were found to have a lower average tumor stage, higher average tumor differentiation, and higher rates of positive chemotherapy responses than those with lower expression levels of SOCS3. SOCS3 promoter methylation was also found to be correlated with chemotherapy responses in these patients. In the prognostic analyses, only SOCS3 expression, but not SOCS3 promoter methylation, was found to be predictive of outcomes in advanced NSCLC patients. We also found that the pro-apoptotic effects of SOCS3 were mediated by the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathways in the lung cancer cells.Conclusions:Currently, there is a lack of reliable biomarkers for predicting the responses of NSCLC patients to chemotherapy. Our results may aid in clinical evaluations of NSCLC patients.

Lupus nephritis (LN) is a common and refractory inflammation of the kidneys caused by systemic lupus erythematosus. Diagnosis and therapies at this stage are inefficient or have severe side effects. In recent years, nanomedicines show great potential for imaging diagnosis and controlled drug release. Herein, we developed a polydopamine (PDA)-based nanocarrier modified with Fe3O4 and Pt nanoparticles and loaded with necrostatin-1 (Nec-1) for the bimodal imaging and therapy of LN. Results demonstrate that Nec-1/PDA@Pt-Fe3O4 nanocarrier exhibits good biocompatibility. Nec-1, as an inhibitor of receptor-interacting protein 1 kinase, can be used to inhibit receptor-interacting protein 1 kinase activity and then reduces inflammation due to LN. Experiments in vitro and in the LN mouse model confirmed that the nanocarrier can reduce neutrophil extracellular traps (NETs) production by RIPK1 and alleviate the progression of inflammation. Previous studies proved that Pt nanoparticles can catalyze H2O2 to produce oxygen. A blood oxygen graph of mouse photoacoustic tomography confirmed that Nec-1/PDA@Pt-Fe3O4 can generate oxygen to fight against the hypoxic microenvironment of LN. PDA and Fe3O4 are used as photographic developers for photoacoustic or magnetic resonance imaging. The preliminary imaging results support Nec-1/PDA@Pt-Fe3O4 potential for photoacoustic/magnetic resonance dual-mode imaging, which can accurately and non-invasively monitor microscopic changes due to diseases. Nec-1/PDA@Pt-Fe3O4 combining these advantages exhibited outstanding performance in LN imaging and therapy. This work offers valuable insights into LN diagnosis and therapy.

OBJECTIVE:To determine the role of gasdermin E (GSDME)-mediated pyroptosis in the pathogenesis and progression of rheumatoid arthritis (RA), and to explore the potential of GSDME as a therapeutic target in RA.METHODS:The expression and activation of caspase 3 and GSDME in the synovium, macrophages, and monocytes of RA patients were determined by immunohistochemistry, immunofluorescence, and Western blot analysis. The correlation of activated GSDME with RA disease activity was evaluated. The pyroptotic ability of monocytes from RA patients was tested, and the effect of tumor necrosis factor (TNF) on caspase 3/GSDME-mediated pyroptosis of monocytes and macrophages was investigated. In addition, collagen-induced arthritis (CIA) was induced in mice lacking Gsdme, and the incidence and severity of arthritis were assessed.RESULTS:Compared to cells from healthy controls, monocytes and synovial macrophages from RA patients showed increased expression of activated caspase 3, GSDME, and the N-terminal fragment of GSDME (GSDME-N). The expression of GSDME-N in monocytes from RA patients correlated positively with disease activity. Monocytes from RA patients with higher GSDME levels were more susceptible to pyroptosis. Furthermore, TNF induced pyroptosis in monocytes and macrophages by activating the caspase 3/GSDME pathway. The use of a caspase 3 inhibitor and silencing of GSDME significantly blocked TNF-induced pyroptosis. Gsdme deficiency effectively alleviated arthritis in a mouse model of CIA.CONCLUSION:These results support the notion of a pathogenic role of GSDME in RA and provide an alternative mechanism for RA pathogenesis involving TNF, which activates GSDME-mediated pyroptosis of monocytes and macrophages in RA. In addition, targeting GSDME might be a potential therapeutic approach for RA.

Background: Neutrophilic inflammation in the airway is a hallmark of bronchiectasis. Neutrophil extracellular traps (NETs) have been reported to play an important role in the occurrence and development of bronchiectasis. Neutrophil side fluorescence is one of the characteristics of neutrophils that can reflect the activation of neutrophils and the formation of NETs. Objective: To explore the relationship between the values of neutrophil side fluorescence (NEUT-SFL) in the peripheral blood of bronchiectasis patients, and the severity of the disease. Methods: 82 patients with bronchiectasis from the Department of Respiratory and Critical Medicine, at the Third Affiliated Hospital of Southern Medical University and were scored with Bronchiectasis Severity Index (BSI) (2019-2021). The clinical data such as the value of NEUT-SFL, neutrophil count, C-reactive protein, and procalcitonin levels were collected and retrospectively analyzed. NEUT-SFL values neutrophil count from 28 healthy subjects were also used to ascertain cut-off values. A single-sample SW test was used to test the normality for the data. The independent sample T-test was performed on the normal data, and Chi-square test and non-parametric test were used for non-normally distributed data and count data. The Pearson correlation analysis method was used for analyzing the correlation between each group of data. Results: Based on the BSI scores, patients were divided into three categories as mild (32%), moderate (29%), and severe (39%). Our results showed that the values of NEUT-SFL were higher in bronchiectasis patients compared to healthy controls. The levels of NEUT-SFL positively correlated with the high BSI scores in patients (P = 0.037, r = 0.23) and negatively correlated with the lung function in these patients (r = -0.35, P = 0.001). The area under the ROC curve was 0.813, the best cut-off was 42.145, indicating that NEUT-SFL values >42.145 can potentially predict the severity of bronchiectasis. Conclusions: The values of NEUT-SFL in the peripheral blood can be used for predicting the severity of bronchiectasis.

OBJECTIVES To determine the role of gasdermin E (GSDME)-mediated pyroptosis in the pathogenesis and progression of rheumatoid arthritis (RA), and explore the potential therapeutic targets for RA. METHODS The expression and activation of caspase3 and GSDME in the synovium, macrophages, and monocytes of RA patients was detected by immunohistochemistry, immunofluorescence, and western blot analysis. The correlation of activated GSDME with RA disease activity was evaluated. The pyroptotic ability of monocytes from RA patients was also tested. In addition, the effect of TNF-α on caspase3/GSDME-mediated pyroptosis of monocytes and macrophages was investigated. Furthermore, mice lacking Gsdme were subjected to collagen-induced arthritis, and the incidence and severity of arthritis was assessed. RESULTS Monocytes and synovial macrophages from RA patients showed increased expression of activated caspase3, GSDME, and GSDME-N. The expression of GSDME-N in monocytes from RA patients correlated positively with the disease activity. Monocytes from RA patients with higher GSDME levels were more susceptible to pyroptosis. Furthermore, TNF-α induced pyroptosis in monocytes and macrophages by activating the caspase3/GSDME pathway. The use of a caspase3 inhibitor and silencing of GSDME significantly blocked TNF-α-induced pyroptosis. Gsdme deficiency effectively alleviated arthritis in a collagen-induced arthritis mice model. CONCLUSIONS These results support a pathogenic role of GSDME in RA and provide an alternative mechanism for RA pathogenesis involving TNF-α, which activates GSDME-mediated pyroptosis of monocytes and macrophages in RA. In addition, targeting GSDME might be a potential therapeutic approach for RA.

目的:探讨CXCR2拮抗剂AZD5069能否通过抑制中性粒细胞迁移治疗类风湿关节炎(RA).方法:将45只雄性DBA/1J小鼠随机分为对照组(n=15)、模型组(n=15)、AZD5069组(n=15);除对照组外,其余两组均给予牛Ⅱ型胶原蛋白+弗氏佐剂建立胶原诱导型关节炎(CIA)模型.观察记录小鼠关节炎评分.二次免疫后第二天给与药物治疗,治疗3周后处死小鼠检测关节病理、中性粒细胞浸润、外周血炎症因子等指标.结果:与对照组相比,模型组小鼠关节炎发病率明显升高(P＜0.05);与模型组相比,AZD5069组小鼠关节炎发病率明显降低(P＜0.05).与对照组相比,模型组小鼠关节炎评分明显升高(P＜0.05);与模型相比,AZD5069组小鼠关节炎评分明显降低(P＜0.05).对照组小鼠踝关节间隙正常,滑膜无增生,无炎症细胞浸润,软骨面光滑无破坏;模型组小鼠踝关节间隙狭窄,滑膜增生明显,可见大量炎症细胞浸润,软骨侵蚀破坏;与模型组相比,AZD5069组小鼠踝关节间隙未见明显狭窄,滑膜未见明显增生,炎症细胞浸润明显减轻,软骨未见明显破坏.与对照组相比,模型组小鼠血清中炎症因子TNF-α、IL-1β的水平明显升高(P＜0.05);与模型组相比,AZD5069组小鼠血清中炎症因子TNF-α、IL-1β的水平明显降低(P＜0.05).与对照组相比,模型组小鼠踝关节中性粒细胞明显增多,可见大量中性粒细胞浸润;与模型组相比,AZD5069组小鼠踝关节中性粒细胞明显减少,中性粒细胞浸润情况明显减轻.结论:CXCR2拮抗剂AZD5069可通过抑制中性粒细胞迁移治疗类风湿关节炎,为开发新型治疗RA药物提供理论基础.

目的 探讨穿心莲内酯(AG)抑制中性粒细胞胞外诱捕网(NETs)的形成及机制,为临床进一步开发应用穿心莲内酯提供理论指导.方法 获取正常健康志愿者外周血,并从中提取中性粒细胞,分为对照组、佛波酯(PMA)组、PMA+AG(12.5、25、50 μmol/L)组、AG(12.5、25、50μmol/L)组,采用PMA诱导中性粒细胞死亡,观察并检测各组中中性粒细胞的NETs形成情况,并检测对应组细胞内的活性氧(ROS)水平.结果 与对照组比较,PMA组中性粒细胞NETs产生的荧光强度值明显增多(P＜0.001),AG(12.5、25、50μmol/L)组荧光强度与对照组比较差异无统计学意义;与PMA组比较,PMA+AG(12.5、25、50 μmol/L)组的荧光强度值均显著降低,并呈浓度依赖性,差异有统计学意义(P＜0.05).与对照组比较,PMA组中性粒细胞ROS产生增多(P＜0.001),AG(12.5、25、50μmol/L)组ROS水平与对照组比较差异无统计学意义;与PMA组比较,PMA+AG(12.5、25、50μmol/L)组ROS水平明显降低,并呈浓度依赖性,差异有统计学意义(P＜0.05).结论 AG可通过降低中性粒细胞ROS的水平抑制NETs产生,提示AG可用于与NETs相关的多种炎症性疾病的治疗.

Background: Activation of the coagulation system has been related to disease activity in some inflammatory diseases. Here, we aimed to investigate the relationship between coagulation function and the disease activity of axial spondyloarthritis (axSpA). Methods: This study retrospectively recruited 144 axSpA patients and 55 healthy controls. The patients were divided into an active group (Bath Ankylosing Spondylitis Disease Activity Index, BASDAI >= 4) and a remission group (BASDAI < 4). The coagulation, inflammatory and clinical parameters were detected. The correlations between these parameters were analyzed with Spearman's correlation analysis. Receiver operating characteristic (ROC) curve analysis was performed to compare the values of these variables in discriminating disease activity. Furthermore, binary logistic regression analysis was used to assess the risk factors for axSpA disease activity. Results: Fibrinogen (FIB) was increased in the axSpA group compared to healthy controls (P < 0.001). Additionally, FIB and D-dimer were higher in the active group than in the remission group (P < 0.05, respectively). FIB and D-dimer were positively correlated with ESR, CRP, BASDAI, Bath Ankylosing Spondylitis Functional Index (BASFI) and Bath Ankylosing Spondylitis Metrology Index (BASMI) (P < 0.05, respectively). The area under the curve (AUC) of FIB was higher than that of ESR, CRP and D-dimer. The optimal cut-off value of FIB was 3.23 g/L, with a specificity of 62.0% and sensitivity of 75.0%. FIB (OR = 4.335, 95% CI: 1.262-14.888, P = 0.020) and BASFI score (OR = 1.878, 95% CI: 1.441-2.448, P < 0.001) were independent risk factors affecting disease activity. Conclusion: Activated coagulation is closely related to the disease activity of axSpA. FIB and D-dimer might be novel indicators for monitoring the disease activity of axSpA.

Neutrophilic inflammation occurs during asthma exacerbation, and especially, in patients with steroid-refractory asthma, but the underlying mechanisms are poorly understood. Recently, a significant accumulation of neutrophil extracellular traps (NETs) in the airways of neutrophilic asthma has been documented, suggesting that NETs may play an important role in the pathogenesis. In this study, we firstly demonstrated that NETs could induce human airway epithelial cell damage in vitro. In a mouse asthmatic model of neutrophil-dominated airway inflammation, we found that NETs were markedly increased in bronchoalveolar lavage (BAL), and the formation of NETs exacerbated the airway inflammation. Additionally, a small-molecule drug necrostatin-1 (Nec-1) shown to inhibit NETs formation was found to alleviate the neutrophil-dominated airway inflammation. Nec-1 reduced total protein concentration, myeloperoxidase activity, and the levels of inflammatory cytokines in BAL. Finally, further experiments proved that the inhibition of Nec-1 on NETs formation might be related to its ability to inhibiting mixed lineage kinase domain-like (MLKL) phosphorylation and perforation. Together, these results document that NETs are closely associated with the pathogenesis of neutrophilic asthma and inhibition of the formation of NETs by Nec-1 may be a new therapeutic strategy to ameliorate neutrophil-dominated airway inflammation.