RÉSUMÉ
OBJECTIVE: To explore the effect of miR-370-3p on LPS triggering, in particular its involvement in disease progression by targeting the TLR4-NLRP3-caspase-1 cellular pyroptosis pathway in macrophages. METHODS: Human macrophage RAW264.7 was divided into 6 groups: control, LPS, LPS + inhibitor-NC, LPS + miR-370-3p inhibitor, LPS + mimics-NC and LPS + miR-370-3p mimics. RT-qPCR was used to detect the expression level of miR-370-3p and analyzed comparatively. CCK-8 and flow cytometry assays were used to detect cell viability and apoptosis. ELISA assay was used to detect the levels of IL-1ß and TNF-α in the supernatant of the cells. The WB assay was used to detect TLR4, NLRP3, Caspase-1 and GSDMD levels. RESULTS: After LPS induction, macrophage miR-370-3p levels decreased, cell viability decreased, and apoptosis increased. At the same time, the levels of TLR4, NLRP3, Caspase-1 and GSDMD increased in the cells, and the levels of IL-1ß and TNF-α increased in the cell supernatant. Compared with the LPS group, the significantly higher expression level of miR-370-3p in the cells of the LPS + miR-370-3p mimics group was accompanied by significantly higher cell viability, significantly lower apoptosis rate, significantly lower levels of TLR4, NLRP3, Caspase-1, and GSDMD in the cells, and significantly lower levels of IL-1ß and TNF-α in the cell supernatant. CONCLUSION: MiR-370-3p may be involved in anti-infective immune responses by targeting and inhibiting the macrophage TLR4-NLRP3-caspase-1 cellular pyroptosis pathway.
Sujet(s)
Caspase-1 , Lipopolysaccharides , Macrophages , microARN , Protéine-3 de la famille des NLR contenant un domaine pyrine , Pyroptose , Récepteur de type Toll-4 , microARN/génétique , microARN/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Récepteur de type Toll-4/métabolisme , Récepteur de type Toll-4/génétique , Lipopolysaccharides/pharmacologie , Macrophages/métabolisme , Macrophages/immunologie , Macrophages/effets des médicaments et des substances chimiques , Humains , Caspase-1/métabolisme , Caspase-1/génétique , Souris , Cellules RAW 264.7 , Animaux , Transduction du signal , Interleukine-1 bêta/métabolisme , Survie cellulaire/effets des médicaments et des substances chimiques , Infections bactériennes/immunologieRÉSUMÉ
OBJECTIVE: To investigate the changes and clinical significance of NOD like receptor protein 3 (NLRP3) inflammasomes and related factors in patients with spinal fractures complicated with acute spinal cord injury (SCI). METHODS: Eighty-six spinal fracture patients complicated with acute SCI admitted to hospital from June 2019 to March 2022 were selected as SCI group, There were 48 males and 38 females, with an average age of (43.48±6.58) years old. And 100 healthy volunteers who underwent physical examination during the same time were selected as control group, including 56 males patients and 44 females patients, with an average age of (45.13±6.43) years old. Peripheral blood mononuclear cell (PBMC) were collected, and the mRNA expressions of NLRP3 and Caspase-1 were detected. Serum was collected and the levels of interleukin (IL)- 1ß, IL-18 were detected. According to Frankel's grade, the SCI group was divided into complete injury patients and incomplete injury patients, and according to the Japanese Orthopedic Society (JOA) grade, the SCI group was divided into good prognosis group and poor prognosis group. The difference of NLRP3, Caspase-1, IL-1ß, IL-18 among groups were compared, the influencing factors for poor prognosis in SCI patients was analyzed by Logistic regression. RESULTS: The mRNA expression levels of NLRP3 (1.41±0.33) and Caspase-1 (1.44±0.35) in PBMC and the levels of IL-1ß(45.34±13.22) pg·ml-1, IL-18(40.95±8.77) pg·ml-1 in serum of SCI group were higher than those of the control group[(1.00±0.19), (1.00±0.16), (16.58±4.24) pg·ml-1, (12.57±3.68) pg·ml-1] (P<0.05). The mRNA expression levels of NLRP3(1.63±0.34) and Caspase-1 (1.67±0.27) in PBMC and the levels of IL-1ß(51.09±11.10) pg·ml-1, IL-18 (47.65±7.93) pg·ml-1 in serum of patients with complete injury in the SCI group were higher than those of patients with incomplete injury [(1.31±0.27), (1.34±0.33), (42.85±13.36) pg·ml-1, (38.05±7.48) pg·ml-1](P<0.05). The mRNA expression levels of NLRP3 (1.66±0.31) and Caspase-1 (1.72±0.31)in PBMC and the levels of IL-1ß(51.21±11.31) pg·ml-1, IL-18 (45.70±7.25) pg·ml-1 in serum, the proportion of complete injury(21 patients), and the proportion of spinal cord edema or bleeding of patients(15 patients) with poor prognosis in the SCI group were higher than those of patients with good prognosis[(1.28±0.26), (1.37±0.36), (42.79±13.25) pg·ml-1ã(38.90±8.63) pg·ml-1, 5ã20 cases](P<0.05). Complete injury and the mRNA expression of NLRP3 in PBMC were the influencing factors for poor prognosis in the SCI group (P<0.05). CONCLUSION: The activation of NLRP3 inflammasomes in patients with spinal fractures complicated with acute SCI is associated with worsening injury and poor prognosis, and NLRP3 expression can serve as a marker for evaluating prognosis.
Sujet(s)
Caspase-1 , Inflammasomes , Interleukine-18 , Interleukine-1 bêta , Protéine-3 de la famille des NLR contenant un domaine pyrine , Traumatismes de la moelle épinière , Fractures du rachis , Humains , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Mâle , Femelle , Traumatismes de la moelle épinière/complications , Traumatismes de la moelle épinière/sang , Adulte , Adulte d'âge moyen , Interleukine-18/sang , Interleukine-1 bêta/sang , Interleukine-1 bêta/génétique , Caspase-1/sang , Fractures du rachis/sang , Fractures du rachis/complications , Agranulocytes/métabolisme , Pronostic , Pertinence cliniqueRÉSUMÉ
Objective: This study aimed to evaluate the role of absent in melanoma 2 (AIM2) inflammasome-mediated pyroptosis in the pathogenesis of acute gouty arthritis (AGA) and asymptomatic hyperuricemia(AHU). Methods: A cohort of 30 AGA patients, 30 AHU individuals, and 30 healthy controls (HC) was assembled. Demographic and biochemical data, along with blood samples, were collected. Serum double-stranded DNA (dsDNA) levels were quantified using a fluorescent assay. Transcriptomic and proteomic analysis of AIM2, Caspase-1, GSDMD, IL-1ß, and IL-18 in peripheral blood mononuclear cells was performed using qRT-PCR and Western blot. Enzyme-linked immunosorbent assay (ELISA) was employed to measure serum IL-1ß and IL-18. Spearman correlation analysis was utilized to assess relationships between variables. Results: Both AGA and AHU groups demonstrated elevated metabolic indicators and serum levels of dsDNA, IL-1ß, and IL-18 compared to the HC group. AGA patients exhibited higher inflammatory markers than the AHU group. In the AGA group, there was a significant increase in the mRNA and protein levels of AIM2, Caspase-1, GSDMD, IL-1ß, and IL-18 (P<0.05 to P<0.001). The AHU group showed higher AIM2, Caspase-1, GSDMD, and IL-18 mRNA levels than the HC group (P<0.001 to P<0.01), with a non-significant increase in AIM2, GSDMD, and IL-1ß proteins (P>0.05). In contrast, Caspase-1 and IL-18 proteins were significantly higher in the AHU group (P<0.05). Notable correlations were observed between AIM2 protein expression and levels of Caspase-1 and GSDMD in both AGA and AHU groups. In the AGA group, AIM2 protein correlated with IL-1ß, but not in the AHU group. The AIM2 protein in the AHU group was positively associated with IL-18, with no such correlation in the AGA group. Conclusion: AIM2 inflammasome may play a role in the inflammatory processes of AGA and AHU and that its activation may be related to the pyroptosis pathway.
Sujet(s)
Goutte articulaire , Protéines de liaison à l'ADN , Hyperuricémie , Inflammasomes , Pyroptose , Humains , Mâle , Inflammasomes/métabolisme , Goutte articulaire/immunologie , Goutte articulaire/sang , Goutte articulaire/métabolisme , Adulte d'âge moyen , Hyperuricémie/sang , Hyperuricémie/immunologie , Femelle , Protéines de liaison à l'ADN/génétique , Protéines de liaison à l'ADN/métabolisme , Adulte , Interleukine-18/sang , Sujet âgé , Études cas-témoins , Marqueurs biologiques/sang , Caspase-1/métabolismeRÉSUMÉ
Cardiovascular complications pose a significant burden in type 2 diabetes mellitus (T2DM), driven by the intricate interplay of chronic hyperglycemia, insulin resistance, and lipid metabolism disturbances. Myocardial ischemia/reperfusion (MI/R) injury during cardiopulmonary bypass (CPB) exacerbates cardiac vulnerability. This study aims to probe the role of Caspase-1-dependent pyroptosis in global ischemia/reperfusion injury among T2DM rats undergoing CPB, elucidating the mechanisms underlying heightened myocardial injury in T2DM. This study established a rat model of T2DM and compared Mean arterial pressure (MAP), heart rate (HR), and hematocrit (Hct) between T2DM and normal rats. Myocardial cell morphology, infarction area, mitochondrial ROS and caspase-1 levels, NLRP3, pro-caspase-1, caspase-1 p10, GSDMD expressions, plasma CK-MB, cTnI, IL-1ß, and IL-18 levels were assessed after reperfusion in both T2DM and normal rats. The role of Caspase-1-dependent pyroptosis in myocardial ischemia/reperfusion injury during CPB in T2DM rats was examined using the caspase-1 inhibitor VX-765 and the ROS scavenger NAC. T2DM rats demonstrated impaired glucose tolerance but stable hemodynamics during CPB, while showing heightened vulnerability to MI/R injury. This was marked by substantial lipid deposition, disrupted myocardial fibers, and intensified cellular apoptosis. The activation of caspase-1-mediated pyroptosis and increased reactive oxygen species (ROS) production further contributed to tissue damage and the ensuing inflammatory response. Notably, myocardial injury was mitigated by inhibiting caspase-1 through VX-765, which also attenuated the inflammatory cascade. Likewise, NAC treatment reduced oxidative stress and partially suppressed ROS-mediated caspase-1 activation, resulting in diminished myocardial injury. This study proved that Caspase-1-dependent pyroptosis significantly contributes to the inflammation and injury stemming from global MI/R in T2DM rats under CPB, which correlate with the surplus ROS generated by oxidative stress during reperfusion.
Sujet(s)
Pontage cardiopulmonaire , Caspase-1 , Diabète de type 2 , Lésion de reperfusion myocardique , Pyroptose , Espèces réactives de l'oxygène , Animaux , Lésion de reperfusion myocardique/métabolisme , Lésion de reperfusion myocardique/étiologie , Lésion de reperfusion myocardique/anatomopathologie , Pontage cardiopulmonaire/effets indésirables , Caspase-1/métabolisme , Diabète de type 2/métabolisme , Diabète de type 2/complications , Rats , Mâle , Espèces réactives de l'oxygène/métabolisme , para-Aminobenzoates/pharmacologie , Rat Sprague-Dawley , Diabète expérimental/métabolisme , Diabète expérimental/complications , Interleukine-18/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , DipeptidesRÉSUMÉ
PROBLEM: Vulvovaginal candidiasis (VVC) is a common mucosal fungal infection, and Candida albicans is the main causative agent. The NLRP3 inflammasome plays an important role in VVC, but the underlying mechanism is unknown. METHOD OF STUDY: Vaginal epithelial cells were divided into three groups: control, C. albicans strain SC5314 (wild-type, WT), and WT+ Matt Cooper Compound 950 (MCC950, a specific NLRP3 inhibitor). After human vaginal epithelial cells were pretreated with 1 µmol/L MCC950 for 2 h, C. albicans (MOI = 1) was cocultured with the human vaginal epithelial cells for 12 h. The cell supernatants were collected, LDH was detected, and the IL-1ß and IL-18 levels were determined by ELISA. The expression of the pyroptosis-related proteins NLRP3, Caspase-1 p20 and GSDMD was measured by Western blotting analysis. The protein expression of the pyroptosis-related N-terminus of GSDMD (GSDMD-N) was detected by immunofluorescence. RESULTS: In this study, we showed that the WT C. albicans strain induced pyroptosis in vaginal epithelial cells, as indicated by the LDH and proinflammatory cytokine levels and the upregulated levels of the pyroptosis-related proteins NLRP3, Caspase-1 p20, and GSDMD-N. MCC950 reversed the changes in the expression of these proteins and proinflammatory cytokines in vaginal epithelial cells. CONCLUSION: C. albicans activated the NLRP3 inflammasome to induce vaginal epithelial cell pyroptosis. MCC950 inhibited the NLRP3 inflammasome, reduced vaginal epithelial cell pyroptosis, and decreased the release of inflammatory cytokines.
Sujet(s)
Candida albicans , Candidose vulvovaginale , Cellules épithéliales , Inflammasomes , Protéine-3 de la famille des NLR contenant un domaine pyrine , Pyroptose , Vagin , Femelle , Humains , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Candidose vulvovaginale/immunologie , Candidose vulvovaginale/microbiologie , Candidose vulvovaginale/métabolisme , Cellules épithéliales/immunologie , Cellules épithéliales/métabolisme , Inflammasomes/métabolisme , Inflammasomes/immunologie , Candida albicans/immunologie , Vagin/microbiologie , Vagin/immunologie , Vagin/anatomopathologie , Interleukine-18/métabolisme , Interleukine-1 bêta/métabolisme , Indènes , Furanes/pharmacologie , Caspase-1/métabolisme , Composés hétérocycliques avec 4 noyaux ou plus/pharmacologie , Protéines de liaison aux phosphates/métabolisme , Cellules cultivées , SulfonamidesRÉSUMÉ
Inflammation through activation of caspase-1, seems to play a role in pulmonary hypertension induced by alveolar hypoxia. Whether alveolar hypoxia induces caspase-1-mediated inflammation and influx of leukocytes in other organs than the lungs, is not known. Our aim was to explore sites of caspase-1-related inflammation in alveolar hypoxia. Wild type (WT) mice were exposed to environmental hypoxia or room-air, and organs were analyzed. Right heart catheterization was performed after 14 days of alveolar hypoxia in WT mice and mice transplanted with WT or caspase-1-/- bone marrow. Hypoxia induced leukocyte accumulation and increased caspase-1 protein in the lungs, not in other organs. WT mice transplanted with WT or caspase-1-/- bone marrow showed no difference in pulmonary leukocyte accumulation or development of pulmonary hypertension after alveolar hypoxia. Caspase-1 and IL-18 were detected in bronchial epithelium in WT mice, and hypoxia induced IL-18 secretion from bronchial epithelial cells. IL-18 stimulation generated IL-6 mRNA in monocytes. Phosphorylated STAT3 was increased in hypoxic lungs, not in other organs. Alveolar hypoxia induces caspase-1 activation and leukocyte accumulation specific to the lungs, not in other organs. Caspase-1 activation and IL-18 secretion from bronchial epithelial cells might initiate hypoxia-induced inflammation, leading to pulmonary hypertension.
Sujet(s)
Caspase-1 , Hypoxie , Inflammasomes , Interleukine-18 , Poumon , Souris de lignée C57BL , Animaux , Mâle , Inflammasomes/métabolisme , Souris , Caspase-1/métabolisme , Caspase-1/génétique , Poumon/métabolisme , Poumon/anatomopathologie , Interleukine-18/métabolisme , Interleukine-18/génétique , Hypoxie/métabolisme , Inflammation/métabolisme , Inflammation/anatomopathologie , Alvéoles pulmonaires/métabolisme , Alvéoles pulmonaires/anatomopathologie , Facteur de transcription STAT-3/métabolisme , Facteur de transcription STAT-3/génétique , Souris knockout , Hypertension pulmonaire/métabolisme , Hypertension pulmonaire/étiologie , Hypertension pulmonaire/anatomopathologieRÉSUMÉ
Bronchopulmonary dysplasia (BPD) is a common serious complication of premature babies. No effective means control it. Hyperoxia damage is one of the important mechanisms of BPD. The reaserach confirmed pyroptosis existed in BPD. Dexmedetomidine is a new, high-specific α2 receptor agonist. Previous research foundation found that dexmedetomidine has a protective effect on BPD. To investigate how dexmedetomidine improves hyperoxic lung injury in neonatal mice by regulating pyroptosis. Neonatal rats were randomly divided into four groups: normal control group, hyperoxic injury group, air plus dexmedetomidine group, and hyperoxia plus dexmedetomidine group. After seven days the lungs of rats in each group were extracted, and the wet-to-dry weight ratio of the lung was measured. The lung injury in rats was observed using hematoxylin-eosin staining. Additionally, the expression and localization of nucleotide-binding oligomerization domain-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), and gasdermin D (GSDMD) proteins were examined in the lungs of rats using immunofluorescence staining. The mRNA levels of NLRP3, ASC, caspase-1, and interleukin 18 (IL-18) in the lungs of rats were determined using real-time PCR. Moreover, the protein levels of NLRP3, ASC, caspase-1/cleaved caspase-1, interleukin 1beta (IL-1ß), IL-18, and tunor necrosis factor alpha (TNF-α) were detected in lungs of rats using Western blot. The extent of mitochondrial damage in lung tissues of each group was observed by transmission electron microscopy. The lung tissue injury of the neonatal rats was significantly improved in the hyperoxia plus dexmedetomidine group compared to the hyperoxic injury group. Furthermore, the expressions of pyroptosis-related proteins such as NLRP3, ASC, cleaved-caspase-1, and GSDMD were significantly decreased, along with the expressions of inflammatory factors in lung tissues. By inhibiting the NLRP3/caspase-1/GSDMD pyroptosis pathway, dexmedetomidine reduces the activation and release of inflammatory factors and provides a protective effect against hyperoxic lung injury in neonatal mice.
Sujet(s)
Animaux nouveau-nés , Dexmédétomidine , Hyperoxie , Lésion pulmonaire , Poumon , Protéine-3 de la famille des NLR contenant un domaine pyrine , Pyroptose , Rat Sprague-Dawley , Animaux , Dexmédétomidine/pharmacologie , Dexmédétomidine/usage thérapeutique , Hyperoxie/métabolisme , Hyperoxie/complications , Hyperoxie/traitement médicamenteux , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Poumon/effets des médicaments et des substances chimiques , Poumon/anatomopathologie , Poumon/métabolisme , Pyroptose/effets des médicaments et des substances chimiques , Lésion pulmonaire/métabolisme , Lésion pulmonaire/prévention et contrôle , Lésion pulmonaire/anatomopathologie , Lésion pulmonaire/traitement médicamenteux , Rats , Protéines de liaison aux phosphates/métabolisme , Protéines adaptatrices de signalisation CARD/métabolisme , Caspase-1/métabolisme , Interleukine-18/métabolisme , Agonistes des récepteurs alpha-2 adrénergiques/pharmacologie , Agonistes des récepteurs alpha-2 adrénergiques/usage thérapeutique , Mâle , GasderminesRÉSUMÉ
Objective: To investigate the effects of zinc oxide nanoparticles (ZnO-NPs) on neutrophil hypoxia and pyroptosis through nucleotide binding of oligomeric domain-like receptor protein 3 (NLRP3) inflammasome, and to analyze the role of pyroptosis on respiratory tract inflammotion induced by ZnO-NPs. Methods: In October 2022, primary cultured neutrophils were obtained from the abdominal aortic blood of SPF adult healthy SD rats. The neutrophils were treated with ZnO-NPs solution (0, 5, 10, 20 µg/ml) at different concentrations, and hypoxia group (5% O(2)) was set up. Hypoxia and reactive oxygen species (ROS) levels were detected by flow cytometry, and the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved Caspase-1 were measured by Western blot. The activity of lactic dehydrogenase (LDH) in the cell supernatant was measured by coloration, and the content of interleukin-1 beta (IL-1ß) in cell culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA) . Results: Compared with the control group, hypoxia and ROS levels of neutrophils in hypoxia group and ZnO-NPs groups were significantly increased (P<0.05), and NLRP3, ASC, cleaved Caspase-1 protein expression levels, LDH activity and IL-1ß content were significantly increased (P<0.05). Compared with hypoxia group, hypoxia and ROS levels of neutrophils in 5 µg/ml and 10 µg/ml ZnO-NPs groups were significantly decreased (P<0.05), NLRP3, ASC, cleaved Caspase-1 protein expression levels, LDH activity, and IL-1ß content were decreased significantly (P<0.05). There were no significant differences in hypoxia, ROS levels, ASC, cleaved Caspase-1 protein expression levels, LDH activity, and IL-1ß content between the 20 µg/ml ZnO-NPs group and the hypoxia group (P>0.05) . Conclusion: ZnO-NPs treatment may activate the NLRP3 inflammasome to induce pyroptosis of neutrophils which may be related to ROS and hypoxia.
Sujet(s)
Caspase-1 , Protéine-3 de la famille des NLR contenant un domaine pyrine , Nanoparticules , Granulocytes neutrophiles , Pyroptose , Rat Sprague-Dawley , Espèces réactives de l'oxygène , Oxyde de zinc , Animaux , Pyroptose/effets des médicaments et des substances chimiques , Rats , Granulocytes neutrophiles/effets des médicaments et des substances chimiques , Granulocytes neutrophiles/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Espèces réactives de l'oxygène/métabolisme , Caspase-1/métabolisme , Interleukine-1 bêta/métabolisme , Inflammasomes/métabolisme , Inflammasomes/effets des médicaments et des substances chimiques , Cellules cultivées , Mâle , Hypoxie/métabolisme , Hypoxie cellulaireRÉSUMÉ
BACKGROUND: Caspase-1 is a crucial component in the inflammasome activation cascade. This study evaluated the potential of serum caspase-1 level as an inflammatory biomarker in patients with adult-onset Still's disease (AOSD). METHODS: The study included 51 consecutive patients diagnosed with AOSD based on the Yamaguchi criteria, 66 patients with rheumatoid arthritis (RA) as disease control, and 36 healthy controls (HCs). Serum caspase-1 concentrations were measured using enzyme-linked immunosorbent assay. The serum 69 cytokine levels were analyzed using a multisuspension cytokine array in patients with AOSD, and a cluster analysis of each cytokine was performed to determine specific molecular networks. RESULTS: Patients with AOSD had significantly increased serum caspase-1 levels versus patients with RA (p < 0.001) and HCs (p < 0.001). Additionally, serum caspase-1 demonstrated significant positive correlations with AOSD disease activity score (Pouchot score, r = 0.59, p < 0.001) and serum ferritin (r = 0.54, p < 0.001). Furthermore, among patients with AOSD, significant correlations existed between serum caspase-1 and inflammatory cytokines, including interleukin-18. Immunoblot analysis detected the cleaved form of caspase-1 (p20) in the serum of untreated patients with AOSD, not in those from patients with inactive AOSD receiving immunosuppressive treatments. CONCLUSIONS: Caspase-1 is a useful biomarker for AOSD diagnosis and monitoring. Caspase-1 activation could be correlated with the inflammatory component of AOSD, specifically through proinflammatory cytokine induction via inflammasome activation cascades.
Sujet(s)
Marqueurs biologiques , Caspase-1 , Maladie de Still débutant à l'âge adulte , Humains , Maladie de Still débutant à l'âge adulte/sang , Maladie de Still débutant à l'âge adulte/diagnostic , Caspase-1/sang , Femelle , Mâle , Adulte , Adulte d'âge moyen , Marqueurs biologiques/sang , Interleukine-18/sang , Études cas-témoins , Cytokines/sang , Polyarthrite rhumatoïde/sang , Polyarthrite rhumatoïde/diagnosticRÉSUMÉ
BACKGROUND: Despite advances in breast carcinoma therapies, drug resistance mechanisms as anti-apoptosis and anti-pyroptosis limit the application of these therapies. This work assesses the immunohistochemical (IHC) expression of Caspase1 and EGFR in breast carcinoma and analyzes their clinicopathological associations as prognostic markers and potential therapeutic targets. Caspase1/EGFR expression patterns are utilized to specify breast carcinoma patients who may benefit from these therapies. METHODS: After reviewing the hematoxylin and eosin-stained slides and the routine breast carcinoma IHC stains (estrogen receptors, progesterone receptors, HER2/NEU, Ki-67) by two pathologists and preparation of tissue microarray blocks, anti-Caspase-1 and EGFR IHC staining was performed using Horseradish Peroxidase (HRP) technique. Intensity and percentage-based scoring was applied dividing the 153 included breast carcinomas into Caspase1-negative and positive expression groups; and EGFR low and overexpression groups. Groups were statistically analyzed in relation to age, tumor size, histological and molecular subtype, grade, nodal status, metastasis/recurrence, TNM stage and Ki-67 proliferation index. Kaplan-Meier's analysis was used to compare disease-free survival (DFS) and overall survival (OS). Combined patterns based on Caspase1 and EGFR expression status were created to stratify patients into prognostic groups. RESULTS: Caspase1 was positive in 54.2% of breast carcinomas and its positivity was significantly associated with smaller tumor size, absence of metastasis/recurrence, luminal A and B molecular subtypes and longer OS (p<0.05). EGFR overexpression was detected in 32.7% of carcinomas and was significantly associated with larger tumor size, TNBLBC and a shorter OS (p<0.05). Caspase1-negative/EGFR-overexpression pattern comprised 14.4% of carcinomas and had the worst prognostic associations including larger tumor size, metastasis/recurrence, TNBLBC subtype and shortest OS (p=0.002, 0.002, 0.004 and ≤0.001 respectively). Conclusions: Combined Caspase1/EGFR IHC expression may provide a tool for selection of patients who benefit from combined EGFR-inhibitors with miR-155-5p down-regulators or photodynamic therapy via induction of apoptosis/pyroptosis in EGFR-overexpression carcinomas through enhanced Caspase1 signaling.
Sujet(s)
Marqueurs biologiques tumoraux , Tumeurs du sein , Caspase-1 , Récepteurs ErbB , Humains , Femelle , Tumeurs du sein/anatomopathologie , Tumeurs du sein/métabolisme , Tumeurs du sein/mortalité , Récepteurs ErbB/métabolisme , Pronostic , Adulte d'âge moyen , Marqueurs biologiques tumoraux/métabolisme , Caspase-1/métabolisme , Taux de survie , Adulte , Études de suivi , Carcinome canalaire du sein/métabolisme , Carcinome canalaire du sein/anatomopathologie , Sujet âgé , Récepteur ErbB-2/métabolisme , Invasion tumorale , Immunohistochimie , Récidive tumorale locale/anatomopathologie , Récidive tumorale locale/métabolismeRÉSUMÉ
Pathogen infection of host cells triggers an inflammatory cell death termed pyroptosis via activation of inflammatory caspases. However, blockade of immune signaling kinases by the Yersinia virulence factor YopJ triggers cell death involving both apoptotic caspase-8 and pyroptotic caspase-1. While caspase-1 is normally activated within inflammasomes, Yersinia-induced caspase-1 activation is independent of known inflammasome components. We report that caspase-8 is an essential initiator, while caspase-1 is an essential amplifier of its own activation through two feed-forward loops involving caspase-1 auto-processing and caspase-1-dependent activation of gasdermin D and NLPR3. Notably, while Yersinia-induced caspase-1 activation and cell death are inflammasome-independent, IL-1ß release requires NLPR3 inflammasome activation. Mechanistically, caspase-8 is rapidly activated within multiple foci throughout the cell, followed by assembly of a canonical inflammasome speck, indicating that caspase-8 and canonical inflammasome complex assemblies are kinetically and spatially distinct. Our findings reveal that functionally interconnected but distinct death complexes mediate pyroptosis and IL-1ß release in response to pathogen blockade of immune signaling.
Sujet(s)
Caspase-1 , Caspase 8 , Inflammasomes , Interleukine-1 bêta , Protéines de liaison aux phosphates , Pyroptose , Transduction du signal , Yersinia , Interleukine-1 bêta/métabolisme , Caspase 8/métabolisme , Animaux , Caspase-1/métabolisme , Inflammasomes/métabolisme , Yersinia/métabolisme , Protéines de liaison aux phosphates/métabolisme , Souris , Humains , Protéines bactériennes/métabolisme , Protéines et peptides de signalisation intracellulaire/métabolisme , Yersinioses/immunologie , Yersinioses/microbiologie , Yersinioses/métabolisme , GasderminesRÉSUMÉ
BACKGROUND: Psoriasis refers to a highly prevalent and immunologically mediated dermatosis with considerable deterioration in life quality. Wogonin, a sort of flavonoid, has been mentioned to elicit protective activities in skin diseases. However, whether Wogonin is implicated in the treatment of psoriasis and its specific mechanisms are not fully understood. AIM: The present work attempted to elaborate the role of Wogonin during the process of psoriasis and to concentrate on the associated action mechanism. METHODS: Cell counting kit-8 (CCK-8) method was initially applied to assay the viability of human keratinocyte HaCaT cells treated by varying concentrations of Wogonin. To mimic psoriasis in vitro, HaCaT cells were exposed to M5 cytokines. CCK-8 and 5-Ethynyl-2'-deoxyuridine assays were adopted for the measurement of cell proliferation. Inflammatory levels were examined with enzyme-linked immunosorbent assay. Immunofluorescence staining tested nucleotide-binding oligomerization domain (NOD)-like receptor family pyrin domain containing 3 (NLRP3) and Caspase-1 expressions. Western blot examined the protein expressions of proliferation-, inflammation-, pyroptosis-associated factors, and NLRP3. RESULTS: Wogonin treatment antagonized the proliferation, inflammatory response, and NLRP3/caspase-1/Gasdermin-D (GSDMD)-mediated pyroptosis in M5-challenged HaCaT cells. Besides, NLRP3 elevation partially abrogated the effects of Wogonin on M5-induced proliferation, inflammatory response, and NLRP3/caspase-1/GSDMD-mediated pyroptosis in HaCaT cells. CONCLUSION: In a word, Wogonin might exert anti-proliferation, anti-inflammatory and anti-pyroptosis activities in M5-induced cell model of psoriasis and the blockade of NLRP3/Caspase-1/GSDMD pathway might be recognized as a potential mechanism underlying the protective mechanism of Wogonin in psoriasis, suggesting Wogonin as a prospective anti-psoriasis drug.
Sujet(s)
Caspase-1 , Prolifération cellulaire , Flavanones , Kératinocytes , Protéine-3 de la famille des NLR contenant un domaine pyrine , Pyroptose , Transduction du signal , Humains , Flavanones/pharmacologie , Pyroptose/effets des médicaments et des substances chimiques , Kératinocytes/effets des médicaments et des substances chimiques , Kératinocytes/métabolisme , Prolifération cellulaire/effets des médicaments et des substances chimiques , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Caspase-1/métabolisme , Transduction du signal/effets des médicaments et des substances chimiques , Protéines et peptides de signalisation intracellulaire/métabolisme , Psoriasis/traitement médicamenteux , Psoriasis/métabolisme , Psoriasis/anatomopathologie , Inflammation/métabolisme , Inflammation/traitement médicamenteux , Cellules HaCaT , Lignée cellulaire , Gasdermines , Protéines de liaison aux phosphatesRÉSUMÉ
Infection with Glaesserella parasuis, the primary pathogen behind Glässer's disease, is often associated with diverse clinical symptoms, including serofibrinous polyserositis, arthritis, and meningitis. Autophagy plays a dual role in bacterial infections, exerting either antagonistic or synergistic effects depending on the nature of the pathogen. Our previous studies have demonstrated that autophagy serves as a defense mechanism, combating inflammation and invasion caused by infection of highly virulent G. parasuis. However, the precise mechanisms remain to be elucidated. Pathogens exhibit distinct interactions with inflammasomes and autophagy processes. Herein, we explored the effect of autophagy on inflammasomes during G. parasuis infection. We found that G. parasuis infection triggers NLRP3-dependent pro-CASP-1-IL-18/IL-1ß processing and maturation pathway, resulting in increased release of IL-1ß and IL-18. Inhibition of autophagy enhances NLRP3 inflammasome activity, whereas stimulation of autophagy restricts it during G. parasuis infection. Furthermore, assembled NLRP3 inflammasomes undergo ubiquitination and recruit the autophagic adaptor, p62, facilitating their sequestration into autophagosomes during G. parasuis infection. These results suggest that the induction of autophagy mitigates inflammation by eliminating overactive NLRP3 inflammasomes during G. parasuis infection. Our research uncovers a mechanism whereby G. parasuis infection initiates inflammatory responses by promoting the assembly of the NLRP3 inflammasomes and activating NLRP3-CASP-1, both of which processes are downregulated by autophagy. This suggests that pharmacological manipulation of autophagy could be a promising approach to modulate G. parasuis-induced inflammatory responses.
Sujet(s)
Autophagie , Caspase-1 , Infections à Haemophilus , Haemophilus parasuis , Inflammasomes , Protéine-3 de la famille des NLR contenant un domaine pyrine , Animaux , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Inflammasomes/immunologie , Inflammasomes/métabolisme , Haemophilus parasuis/immunologie , Haemophilus parasuis/pathogénicité , Haemophilus parasuis/génétique , Caspase-1/métabolisme , Caspase-1/génétique , Infections à Haemophilus/médecine vétérinaire , Infections à Haemophilus/immunologie , Infections à Haemophilus/microbiologie , Suidae , Interleukine-18/métabolisme , Interleukine-18/génétique , Interleukine-1 bêta/métabolisme , Interleukine-1 bêta/génétique , Maladies des porcs/microbiologie , Maladies des porcs/immunologie , SourisRÉSUMÉ
BACKGROUND: Our previous studies have demonstrated a strong relationship betweenCutibacterium acnes(C. acnes), oxidative stress, and acne inflammation. Syringic acid (SA) is a plant widely used for its antimicrobial, anti-inflammatory, and antioxidant activities, but lacking data on acne. This study aims to investigate the effect and mechanism of SA on acne inflammation induced by C. acnes in vitro and in vivo. METHODS: After using the SA to expose HaCaT keratinocytes, we reevaluated the effect of the SA on cell viability, cell apoptosis, ROS, CAT, SOD, and other inflammatory variables in the heat-killed C. acnes-treated HaCaT cells. Next, to induce mice with acne inflammation, ICR mice were given an intradermal injection of live C. acnes into their right ears. The effect of SA on this inflammation was then examined. Moreover, we explored the mechanism of SA on PPARγ/Nrf2 and NLRP3/caspase-1/IL-1ß pathways by ELISA, immunofluorescence microscopy, and western blot assay. RESULTS: Heat-killed C. acnes triggered remarkable cell apoptosis, ROS production, interleukin (IL)-1ß, IL-18, IL-6, and TNF-α release, reduced SOD and CAT activity, and upregulated the expression of proteins in HaCaT cells, including up-regulating IL-1ß, PPARγ, Nrf2, HO-1, NQO1, NLRP3, and caspase-1, whereas SA inhibited these effects by partially impairing PPARγ activation. In addition, PPARγ silencing decreased C. acnes-induced IL-1ß secretion and the production of intracellular ROS, down-regulating the expression of Nrf2. Nrf2 activator (SFN) enhanced anti-inflammatory activity through antioxidant mechanisms, boosting intracellular ROS production, reducing SOD and CAT activity, and promoting the increase in ROS, HO-1, NQO1, and IL-1ß levels, while PPARγ inhibitor (GW662) effectively inhibited this effect in heat-killed C. acnes-treated cells. Finally, SA also exhibited notable improvements in ear redness, swelling, and the expression of PPARγ, NLRP3, and IL-1ß in vivo. CONCLUSIONS: SA inhibited C. acnes-induced inflammation via regulating the NLRP3/caspase-1/IL-1ß signaling axis by activating the PPARγ/Nrf2-antioxidant pathway, suggesting a new treatment possibility for acne vulgaris.
Sujet(s)
Acné juvénile , Anti-inflammatoires , Caspase-1 , Acide gallique , Interleukine-1 bêta , Kératinocytes , Souris de lignée ICR , Facteur-2 apparenté à NF-E2 , Protéine-3 de la famille des NLR contenant un domaine pyrine , Récepteur PPAR gamma , Transduction du signal , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Humains , Facteur-2 apparenté à NF-E2/métabolisme , Récepteur PPAR gamma/métabolisme , Animaux , Caspase-1/métabolisme , Kératinocytes/effets des médicaments et des substances chimiques , Kératinocytes/immunologie , Transduction du signal/effets des médicaments et des substances chimiques , Interleukine-1 bêta/métabolisme , Anti-inflammatoires/pharmacologie , Anti-inflammatoires/usage thérapeutique , Acné juvénile/traitement médicamenteux , Acné juvénile/microbiologie , Acné juvénile/immunologie , Souris , Acide gallique/analogues et dérivés , Acide gallique/pharmacologie , Acide gallique/usage thérapeutique , Cellules HaCaT , Antioxydants/pharmacologie , Antioxydants/usage thérapeutique , Inflammation/traitement médicamenteux , Apoptose/effets des médicaments et des substances chimiques , Lignée cellulaire , Propionibacterium acnesRÉSUMÉ
AIMS: Depression is a potentially fatal illness affecting millions of individuals worldwide, across all age groups. Neuroinflammation is a key factor in depression development. Paclitaxel (PXL), a well-known chemotherapeutic agent has been used as therapy for several types of cancer. This study aims to evaluate the ameliorative effect of low-dose PXL against lipopolysaccharide (LPS)-induced depression in rats. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were administrated a single dose of LPS (5 mg/kg, i.p.); 2 h later, rats received PXL (0.3 mg/kg, i.p. three times/week) for one week. KEY FINDINGS: Low-dose PXL alleviated LPS-induced depressive-like behavior in rats as evidenced by significantly improving behavioral changes in both forced swim test (FST) and open field test (OFT), successfully mitigated depletion of monoamines (serotonin, norepinephrine, and dopamine), in addition to markedly decreasing lipid peroxidation with antioxidant levels elevation in brain tissues. Low-dose PXL substantially decreased inflammation triggered by LPS in brain tissue via repressing the expression of NLRP3 and its downstream markers level, caspase-1 and IL-1ß jointly with a corresponding decrease in proinflammatory cytokine levels (TNF-α). Furthermore, low-dose PXL remarkably down-regulated Sphk1/S1P signaling pathway. Concurrent with these biochemical findings, there was a noticeable improvement in the brain tissue's histological changes. SIGNIFICANCE: These findings prove the role of low-dose PXL in treatment of LPS-induced neuroinflammation and depressive-like behavior through their anti-depressant, antioxidant and anti-inflammatory actions. The suggested molecular mechanism may entail focusing the interconnection among Sphk1/S1P, and NLRP3/caspase-1/IL-1ß signaling pathways. Hence PXL could be used as a novel treatment against LPS-induced depression.
Sujet(s)
Caspase-1 , Dépression , Interleukine-1 bêta , Lipopolysaccharides , Facteur de transcription NF-kappa B , Protéine-3 de la famille des NLR contenant un domaine pyrine , Paclitaxel , Rat Sprague-Dawley , Transduction du signal , Animaux , Mâle , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Lipopolysaccharides/toxicité , Transduction du signal/effets des médicaments et des substances chimiques , Dépression/traitement médicamenteux , Dépression/induit chimiquement , Dépression/métabolisme , Caspase-1/métabolisme , Interleukine-1 bêta/métabolisme , Rats , Facteur de transcription NF-kappa B/métabolisme , Paclitaxel/toxicité , Phosphotransferases (Alcohol Group Acceptor)/métabolisme , Maladies neuro-inflammatoires/traitement médicamenteux , Maladies neuro-inflammatoires/induit chimiquement , Maladies neuro-inflammatoires/métabolisme , Comportement animal/effets des médicaments et des substances chimiques , Antidépresseurs/pharmacologie , Antidépresseurs/usage thérapeutique , Anti-inflammatoires/pharmacologieRÉSUMÉ
BACKGROUND/AIM: There is lack of studies assessing the correlation between pain scales and acute phase immune response (APR) following surgery. The purpose of this work was to assess the correlation between cysteine protease caspase-1 (Casp1) blood levels and two pain scales in a cohort of 56 midline laparotomy (MLa) patients and to assess their link with other cytokines (CYTs). PATIENTS AND METHODS: Blood levels of Casp1 and other CYTs (IL-18, IL-18BP, IL-1ra, IL-6, IL-8, IL-10, IL-1ß) were measured before operation and following surgery in patients with MLa. Pain levels were assessed using the Numerical Rating Scale (NRS) and Brief Pain Inventory (BPI) scale, both preoperatively and postoperatively. RESULTS: Casp1 blood levels showed an increasing trend at postoperative day 1 (POP1) and this increase was almost significant in a linear mixed effect model (LME) analysis (p=0.06). Additionally, Casp1 blood levels were higher in patients with cancer than those with benign disease and correlated with IL-18 blood levels (r=0.24, p=0.007). Furthermore, Casp1 blood levels correlated with BPIsev (severity) score values in MLa patients (r=-0.49, p=0.048). A significant correlation was also observed between Casp1 blood levels and NRS scores in patients with MLa. CONCLUSION: This is the first report to evaluate two pain surveys (NRS and BPI) in MLa patients in relation to blood levels of Casp1 and eight CYTs. This analysis is important in confirming the significant correlation between NRS and BPI pain scales and Casp1 blood levels. Our study is also the first to demonstrate that adequate postoperative analgesia in patients with MLa provides better functional ability and improved patient satisfaction.
Sujet(s)
Caspase-1 , Laparotomie , Tumeurs , Mesure de la douleur , Douleur postopératoire , Humains , Femelle , Laparotomie/effets indésirables , Mâle , Caspase-1/sang , Adulte d'âge moyen , Études prospectives , Mesure de la douleur/méthodes , Tumeurs/chirurgie , Tumeurs/sang , Douleur postopératoire/sang , Douleur postopératoire/étiologie , Sujet âgé , Adulte , Cytokines/sangRÉSUMÉ
PURPOSE: In developed countries, endometrial cancer (EC) is the most prevalent gynecological cancer and its occurrence is associated with chronic inflammation. ATP5F1D is a subunit of ATP synthase (complex V), as well as the important component of mitochondrial electron transport chain (ETC). ETC play compelling roles in carcinogenesis. To date, little is known about the role of ATP5F1D in EC. METHODS: ATP5F1D expression was identified in EC tissues and EC cell lines. We evaluated the influence of ATP5F1D on clinical features and prognosis based on TCGA database. The effects of ATP5F1D in EC malignant progression by applying loss-of-function assays in KLE and Ishikawa cell lines were detected by EdU, CCK-8, wound healing, Transwell, and flow cytometry assays. Additionally, electron microscope, LDH release, ELISA, mitochondrial ROS measurement, and Immunofluorescence were performed to demonstrate ATP5F1D can affect the pyroptosis of EC. To observe the anti-tumor effect on ATP5F1D silencing, we established an in vivo human endometrial tumor model using nude mice. RESULTS: ATP5F1D expression was significantly upregulated in EC and was associated with favorable prognosis. ATP5F1D knockdown inhibited the proliferation, invasion, and migration of EC cells. Similarly, in nude mice, ATP5F1D knockdown suppressed the growth EC cells. Knocking down ATP5F1D lead to decrease the production of mitochondrial ROS (mtROS) and inhibited pyroptosis of EC cells. CONCLUSION: Downregulation of ATP5F1D as a new therapeutic strategy that could mediate pyroptosis via suppressing mtROS/NLRP3/caspase-1/GSDMD pathway to inhibit EC progression.
Sujet(s)
Caspase-1 , Tumeurs de l'endomètre , Souris nude , Mitochondrial Proton-Translocating ATPases , Protéine-3 de la famille des NLR contenant un domaine pyrine , Pyroptose , Humains , Femelle , Tumeurs de l'endomètre/anatomopathologie , Tumeurs de l'endomètre/métabolisme , Tumeurs de l'endomètre/génétique , Animaux , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Lignée cellulaire tumorale , Caspase-1/métabolisme , Mitochondrial Proton-Translocating ATPases/métabolisme , Mitochondrial Proton-Translocating ATPases/génétique , Souris , Régulation négative , Évolution de la maladie , Souris de lignée BALB C , Mitochondries/métabolisme , Transduction du signal , Espèces réactives de l'oxygène/métabolisme , Régulation de l'expression des gènes tumoraux , Gasdermines , Protéines de liaison aux phosphatesRÉSUMÉ
ETHNOPHARMACOLOGICAL RELEVANCE: Trifolium alexandrinum L. (TA), has traditionally been used in folk medicine for its anti-inflammatory properties against hyperuricemia and gout. However, the specific mechanisms of action of TA have not been thoroughly studied. AIM OF THE WORK: This study aimed to evaluate the protective effects of irradiated (TR25) and non-irradiated (TR0) Trifolium alexandrinum L. aqueous extract (TAAE), along with two isolated compounds, caffeine (CAF) and saponin (SAP), in a rat model of acute gouty arthritis (GA). MATERIALS AND METHODS: The GA model was established by injecting a monosodium urate (MSU) suspension into the knee joint. Synovial tissue pathology was assessed, and levels of TNF-α, IL-6, IL-1ß, NF-κB, mTOR, AKT1, PI3K, NLRP3, and ASC were measured by ELISA. mRNA expression of ERK1, JNK, and p-38 MAPK was detected using qRT-PCR, and Caspase-1 protein expression was assessed by immunohistochemical analysis. Knee swelling, uric acid levels, liver and kidney function, and oxidative stress markers were also evaluated. RESULTS: TAAE analysis identified 170 compounds, with 73 successfully identified using LC-HR-MS/MS, including caffeine citrate and theasapogenol B glycoside as the main constituents. The studied materials demonstrated significant protective effects against GA. TR25 administration significantly mitigated knee joint circumference compared to other treatments. It demonstrated potential in alleviating hyperuricemia, renal and hepatic impairments induced by MSU crystals. TR25 also alleviated oxidative stress and reduced levels of IL1ß, IL-6, TNF-α, and NF-κB. Weak Caspase-1 immune-positive staining was observed in the TR25 group. TR25 decreased NLRP3 and ASC expression, reducing inflammatory cytokine levels in GA. It effectively inhibited the PI3K, AKT, and mTOR signaling pathways, promoting autophagy. Additionally, TR25 suppressed ERK1, JNK, and p-38 MAPK gene expression in synovial tissue. These effects were attributed to various components in TAAE, such as flavonoids, phenolic acids, tannins, alkaloids, and triterpenes. CONCLUSION: Importantly, irradiation (25 KGy) enhanced the antioxidant effects and phtchemical contents of TAAE. Additionally, TR0, TR25, CAF, and SAP exhibited promising protective effects against GA, suggesting their therapeutic potential for managing this condition. These effects were likely mediated through modulation of the NLRP3/ASC/Caspase-1 and ERK/JNK/p-38 MAPK signaling pathways, as well as regulation of the PI3K/AKT/mTOR pathway. Further research is warranted to fully elucidate the underlying mechanisms and optimize their clinical applications.
Sujet(s)
Goutte articulaire , Facteur de transcription NF-kappa B , Protéine-3 de la famille des NLR contenant un domaine pyrine , Extraits de plantes , Animaux , Goutte articulaire/traitement médicamenteux , Goutte articulaire/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Extraits de plantes/pharmacologie , Extraits de plantes/composition chimique , Extraits de plantes/usage thérapeutique , Mâle , Facteur de transcription NF-kappa B/métabolisme , Rats , Rat Sprague-Dawley , Caspase-1/métabolisme , Transduction du signal/effets des médicaments et des substances chimiques , Anti-inflammatoires/pharmacologie , Système de signalisation des MAP kinases/effets des médicaments et des substances chimiques , Acide uriqueRÉSUMÉ
Psoraleae Fructus (PF, Psoralea corylifolia L.), a traditional medicine with a long history of application, is widely used clinically for the treatment of various diseases. However, the reports of PF-related adverse reactions, such as hepatotoxicity, phototoxic dermatitis, and allergy, are increasing year by year, with liver injury being the mostly common. Our previous studies have demonstrated that PF and its preparations can cause liver injury in lipopolysaccharide (LPS)-mediated susceptibility mouse model, but the mechanism of PF-related liver injury is unclear. In this study, we showed that PF and bavachinin, a major component of PF, can directly induce the expression of caspase-1 and interleukin-1ß (IL-1ß), indicating that PF and bavachinin can directly triggered the activation of inflammasome. Furthermore, pretreatment with NLR family pyrin domain-containing 3 (NLRP3), NLR family CARD domain containing 4 (NLRC4) or absent in melanoma 2 (AIM2) inflammasome inhibitors, containing MCC950, ODN TTAGGG (ODN) and carnosol, all significantly reversed bavachinin-induced inflammasome activation. Mechanistically, bavachinin dose-dependently promote Gasdermin D (GSDMD) post-shear activation and then induce mitochondrial reactive oxygen species (mtROS) production and this effect is markedly inhibited by pretreatment with N-Acetylcysteine amide (NAC). In addition, combination treatment of LPS and bavachinin significantly induced liver injury in mice, but not LPS or bavachinin alone, and transcriptome analysis further validated these results. Thus, PF and bavachinin can induce the activation of inflammasome by promoting GSDMD cleavage and cause hepatotoxicity in mice. Therefore, PF, bavachinin, and PF-related preparations should be avoided in patients with inflammasome activation-associated diseases.
Sujet(s)
Inflammasomes , Protéines de liaison aux phosphates , Psoralea , Pyroptose , Animaux , Pyroptose/effets des médicaments et des substances chimiques , Souris , Protéines de liaison aux phosphates/métabolisme , Protéines de liaison aux phosphates/génétique , Psoralea/composition chimique , Inflammasomes/métabolisme , Protéines et peptides de signalisation intracellulaire/métabolisme , Mâle , Lésions hépatiques dues aux substances/métabolisme , Lésions hépatiques dues aux substances/anatomopathologie , Souris de lignée C57BL , Lipopolysaccharides/toxicité , Foie/effets des médicaments et des substances chimiques , Foie/métabolisme , Foie/anatomopathologie , Flavonoïdes/pharmacologie , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Caspase-1/métabolisme , Interleukine-1 bêta/métabolisme , GasderminesRÉSUMÉ
AIMS: Adipose-derived stem cell (ADSC)-derived exosomes have been recognized for their neuroprotective effects in various neurological diseases. This study investigates the potential neuroprotective effects of ADSC-derived exosomes in sepsis-associated encephalopathy (SAE). METHODS: Behavioral cognitive functions were evaluated using the open field test, Y-maze test, and novel object recognition test. Brain activity was assessed through functional magnetic resonance imaging (fMRI). Pyroptosis was measured using immunofluorescence staining and western blotting. RESULTS: Our findings indicate that ADSC-derived exosomes mitigate cognitive impairment, improve survival rates, and prevent weight loss in SAE mice. Additionally, exosomes protect hippocampal function in SAE mice, as demonstrated by fMRI evaluations. Furthermore, SAE mice exhibit neuronal damage and infiltration of inflammatory cells in the hippocampus, conditions which are reversed by exosome treatment. Moreover, our study highlights the downstream regulatory role of the NLRP3/caspase-1/GSDMD signaling pathway as a crucial mechanism in alleviating hippocampal inflammation. CONCLUSION: ADSC-derived exosomes confer neuroprotection in SAE models by mediating the NLRP3/caspase-1/GSDMD pathway, thereby ameliorating cognitive impairment.