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Inflammation is a key response to a variety of stimuli, and inflammatory molecules such as cytokines play a central role in the pathogenesis of many diseases, including bone and joint diseases. Proinflammatory cytokines are mainly produced by immune cells, which mediate inflammation and innate immune responses. Inflammasomes are newly recognized as important players in innate immunity. The most typical one is the NOD-like receptor protein 3 (NLRP3) inflammasome. NLRP3 detects exogenous pathogenic invasion and endogenous cell damage and responds by forming the NLRP3 inflammasome. The three main components of the NLRP3 inflammasome are NLRP3 (capturing danger signals and recruits downstream molecules), caspase-1 (inducing the maturation of cytokines IL-18 and IL-18 to mediate the release and focal death of cytokines) and apoptosis-related spot-like protein containing caspase recruitment domain (ASC, which connects NLRP3 and caspase-1). In addition, proinflammatory cytokines accelerate bone absorption and cartilage destruction, leading to bone and joint tissue destruction. Therefore, proinflammatory cytokines are involved in regulating the pathogenesis of bone and joint diseases. This paper reviewed the research progress on the activation and function of NLRP3 inflammatory corpuscles in bone and joint diseases in recent years. [ABSTRACT FROM AUTHOR]

Objective To investigate the role of sodium-hyaluronate-modified calcium peroxide nanoparticles (SH-CaO2 NPs) in inducing pyroptosis in human gastric cancer cells and its possible mechanisms. Methods Transmission electron microscopy (TEM), X-ray diffraction (XRD), infrared spectroscopy, and zeta potential test were used to confirm the synthesis of SH-CaO2 NPs. Cell scratch assay and CCK-8 assay were employed to observe the impacts of SH-CaO2 NPs on the migration and proliferation of human gastric cancer cell line HGC-27. The generation of reactive oxygen species (ROS) was observed with confocal laser scanning microscopy (CLSM) and quantified with flow cytometry in the cells after SH-CaO2 NPs treatment or with pretreatment with ROS inhibitor NAC. Furthermore, the effects of pretreatment of NLRP3 inhibitor (MCC950) and Caspase-1 inhibitor (VX765) on the proliferative activity and on expression of their own and their downstream GSDMD in HGC-27 cells were also investigated with CCK-8 assay, immunofluorescence assay and Western blotting. Results TEM images, XRD, infrared spectroscopy, and zeta potential test confirmed the successful preparation of SH-CaO2 NPs. Cell scratch assay and CCK-8 assay showed that application of SH-CaO2 NPs for 24 h significantly inhibited the proliferation of HGC-27 cells (P < 0.001), while, CLSM and flow cytometry indicated the treatment also promoted the production of ROS (P < 0.001). Pretreatment of ROS inhibitor NAC resulted in up-regulation of NLRP3, and increased expression levels of cleaved Caspase-1 and N-terminal fragment of GSDMD (P < 0.001), while pretreatment of both NLRP3 inhibitor and Caspase-1 inhibitor could reverse the process. Conclusion SH-CaO2 NPs inhibit cell viability of human gastric cancer, which may mediate the inflammatory response and pyroptosis by activating the ROS/NLRP3/Caspase-1/GSDMD signaling pathway.

Introduction: NOD-like receptor protein 3 (NLRP3) is implicated in chronic obstructive pulmonary disease (COPD) pathogenesis. Here, we explored the role of NLRP3 in cigarette smoke (CS)-induced airway inflammation in COPD. Material and methods: NLRP3 expression level was assessed with the microarray data in GEO datasets and validated in serum by ELISA from a case-control cohort. Male C57BL/6J mice were randomly divided into: saline, CS, MCC950 (a specific NLRP3 inhibitor, 10 mg/kg) and CS + MCC950 (5 mg/kg and 10 mg/kg) groups (n = 5 per group). All mice were exposed to CS or air for 4 weeks. Then, broncho-alveolar lavage (BAL) fluid and lung tissues were collected for cell counting, ELISA, HE staining and RNA sequencing with validation by real-time qPCR. Results: Compared to non-smokers, NLRP3 expression was significantly elevated in the lung tissues and sera of COPD smokers. CS remarkably induced airway inflammation in mice, characterized by an increase of inflammatory cells and proinflammatory cytokines in BAL fluid and HE inflammatory score, which were ameliorated by MCC950 treatment dose-dependently. Subsequently, 84 candidate genes were selected following RNA sequencing, and five hub genes (Mmp9, IL-1a, Cxcr2, Cxcl10, Ccr1) were then identified by PPI and MCODE analyses, which were confirmed by real-time qPCR. GO and KEGG analysis suggested that the five genes were enriched in a complicated network of inflammatory processes and signaling pathways. Conclusions: NLRP3 expression is elevated in lungs and sera of COPD smokers. Inhibition of NLRP3 significantly attenuates CS-induced airway inflammation in mice via inactivation of multiple hub genes and their related inflammatory processes and signaling pathways. [ABSTRACT FROM AUTHOR]

Excessive inflammatory response and oxidative stress (OS) play an important role in the pathogenesis of spinal cord injury (SCI). Balance of inflammation and prevention of OS have been considered an effective strategy for the treatment of SCI. Hyaluronan and proteoglycan link protein 1 (HAPLN1), also known as cartilage link protein, has displayed a wide range of biological and physiological functions in different types of tissues and cells. However, whether HAPLN1 regulates inflammation and OS during SCI is unknown. Therefore, we aimed to examine whether HAPLN1 can have a protective effect on SCI. In this study, both in vitro and in vivo SCI models were established. Nissl staining and terminal deoxynucleotidyl transferase dUTP nick end labeling staining assays were used. Western blotting and enzyme‐linked immunosorbent assay were employed to assess the expression of proteins. Our results demonstrate that the administration of HAPLN1 promoted the recovery of motor neurons after SCI by increasing the Basso mouse scale score, increasing the numbers of motor neurons, and preventing apoptosis of spinal cord cells. Additionally, HAPLN1 mitigated OS in spinal cord tissue after SCI by increasing the content of superoxide dismutase SOD and the activity of glutathione peroxidase but reducing the levels of malondialdehyde. Importantly, we found that HAPLN1 stimulated the activation of the nuclear factor erythroid 2‐related factor 2 (Nrf2)/antioxidant response element (ARE) pathway and stimulated the expression of heme oxygenase‐1 and nicotinamide adenine dinucleotide phosphate quinone oxidoreductase‐1, which mediated the attenuation of HAPLN1 in activation of the NOD‐like receptor protein 3 (NLRP3) inflammasome by reducing the levels of NLRP3, apoptosis‐associated speck‐like protein containing a caspase recruitment domain (ASC), caspase‐1, and interleukin‐1β. Correspondingly, in vitro experiments show that the presence of HAPLN1 suppressed the NLRP3 inflammasome and prevented cell injury against H2O2 in PC12 cells. These effects were mediated by the Nrf2/ARE pathway, and inhibition of Nrf2 with ML385 abolished the beneficial effects of HAPLN1. Based on these findings, we conclude that HAPLN1 inhibits the NLRP3 inflammasome through the stimulation of the Nrf2/ARE pathway, thereby suppressing neuroinflammation, enhancing motor neuronal survival, and improving the recovery of nerve function after SCI. [ABSTRACT FROM AUTHOR]

Activation of the NOD-like receptor protein 3 (NLRP3) inflammasome signaling pathway is an important mechanism underlying myocardial pyroptosis and plays an important role in inflammatory damage to myocardial tissue in patients with cardiovascular diseases (CVDs), such as diabetic cardiomyopathy, ischemia/reperfusion injury, myocardial infarction, heart failure and hypertension. Noncoding RNAs (ncRNAs) are important regulatory factors. Many Chinese medicine (CM) compounds, including their effective components, can regulate pyroptosis and exert myocardium-protecting effects. The mechanisms underlying this protection include inhibition of inflammasome protein expression, Toll-like receptor 4–NF-κB signal pathway activation, oxidative stress, endoplasmic reticulum stress (ERS), and mixed lineage kinase 3 expression and the regulation of silent information regulator 1. The NLRP3 protein is an important regulatory target for CVD prevention and treatment with CM. Exploring the effects of the interventions mediated by CM and the related mechanisms provides new ideas and perspectives for CVD prevention and treatment. [ABSTRACT FROM AUTHOR]

Objective: To investigate the protective effect of dexmedetomidine (DEX) on myocardial ischemia-reperfusion injury in diabetic mice and its possible molecular mechanism. Methods: Sixty 8-week-old SPF C57BL mice were fed with high fat for 6 weeks. At the 7th week, the type 2 diabetes model was established by intraperitoneal injection of streptozotocin (STZ) 45 mg/kg/d once a day for5 days. After modeling, the mice were randomly divided into sham operation group (sham group), ischemia-reperfusion group (I/R group), hypoxia-inducible factor-1α (HIF-1α) inhibitor 2ME2 group (2ME2 group), DEX group and DEX+2ME2 group (DM group), with 12mice in each group. In sham group, the skin was only cut and sutured. In the other four groups, the left anterior descending coronary artery was ligated through thoracotomy, and the ligating wire was released after 60 minutes of ischemia, followed by 120 minutes of reperfusion to establish the ischemia-reperfusion injury model. Blood samples were collected from the abdominal aorta at 120 min of reperfusion for determination of serum concentrations of troponin I (cTnI), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α)by ELISA. Then the mice were sacrificed and left ventricles were isolated for observation of morphology and structure of myocardium by HE staining. The expression of HIF-1α and NOD-like receptor protein 3 (NLRP3) was detected by Western blot. Cardiac function was assessed by echocardiography at 24 h of reperfusion. Results: Compared with sham group, the concentrations of cTnI, IL-1β and TNF-α were significantly increased, myocardial tissue structure disorder, myocardial fiber rupture, myocardial cell swelling, inflammatory cell infiltration were significantly increased, the expression of NLRP3 in myocardial tissue was significantly increased, and stroke volume (SV), ejection fraction (EF) % and fractional shortening (FS) % were significantly decreased in I/R, 2ME2, DEX and DM groups (P＜0.05).Compared with I/R group, the expression of HIF-1α was significantly increased, the expression of NLRP3 was significantly decreased, the concentrations of cTnI, IL-1β and TNF-α were significantly decreased, myocardial tissue structure was significantly improved, inflammatory cell infiltration was significantly decreased, SV, EF and FS were significantly increased in DEX and DM groups (P＜0.05). Compared with the DEX group, the expression of HIF-1α was significantly decreased, the expression of NLRP3 was significantly increased, the concentrations of cTnI, IL-1β and TNF-α were significantly increased, myocardial tissue structure disorder, myocardial fiber rupture, myocardial cell swelling, inflammatory cell infiltration were significantly increased, SV, EF and FS were significantly decreased in the DM group (P＜0.05). Conclusions: DEX can alleviate myocardial I/R injury in diabetic mice by up-regulating HIF-1α, inhibiting the activation of NLRP3 inflammasome and reducing inflammatory response. [ABSTRACT FROM AUTHOR]

Objective To observe the preventive and therapeutic effect of intraperitoneal and subcutaneous injection of allopregnanolone (Allo) on postoperative delirium (POD) in aged male mice and to explore its possible mechanism. Methods Ninety-six 6-month-old male C57BL/6J mice were randomly divided into groups A, B, C and control group, with 24 in each. Mice in the control group were fed normally, and mice in the groups A, B and C underwent internal fixation of tibial fracture of right lower limb (POD animal model preparation method is commonly used). Mice in the group A were intraperitoneally injected with 8 mg/kg allopregnenolone at 1 h after surgery, and then were subcutaneously injected with 8 mg/kg allopregnenolone at 6, 24 and 48 h after surgery, respectively. Mice in the group B were intraperitoneally injected with 2-hydroxypropyl-β-cyclodextrin at 1 h after surgery, and subcutaneously injected with 2-hydroxypropyl-β-cyclodextrin at 6, 24 and 48 h after surgery, respectively. Mice in the group C were not given any drugs after surgery. Morris water maze test was used to evaluate the spatial learning and memory ability of mice before surgery and at the 1, 3 and 7 days after surgery, respectively. After the test, six mice were randomly killed in each group. Hippocampal tissues were taken to detect inflammatory factors and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK), nuclear factor κB (NF-κB), and NOD-like receptor protein 3 (NLRP3) pathway-related proteins in the hippocampal tissues of mice in each group. Results Compared with the control group, the escape latency of mice in group C was longer and the number of platform crossing was smaller on the 1st, 3rd and 7th days after operation (all P<0. 05). Compared with the group C, the escape latency of mice in the group A was shorter and the number of platform crossing was lager on day 1, 3 and 7 after operation (all P<0. 05). Compared with the control group, the expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), p-p38MAPK, NF-κB, and NLRP3 in the hippocampal tissues of group C were higher on day 1, 3 and 7 after surgery (all P<0. 05). Compared with the group C, the expression levels of TNF-α, IL-1β, p-p38MAPK, NF-κB and NLRP3 in the hippocampal tissues of the group A were lower on day 1, 3 and 7 after surgery (all P<0. 05). Conclusion Intraperitoneal and subcutaneous injection of allopregnenolone can reduce POD after internal fixation of tibial fracture of right lower limb in aged male mice probably by inhibiting the expression levels of TNF-α, IL-1β, p-p38MAPK, NF-κB, and NLRP3 in the hippocampal tissues. [ABSTRACT FROM AUTHOR]

It is well-established that cardiovascular disease and depression are highly comorbid. This study aimed to assess the possible role of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the high-sensitivity C-reactive protein (hsCRP) in patients with incident myocardial infarction in the presence or absence of depression. Sixty-eight consecutive patients with incident ST-elevation myocardial infarction and twenty healthy subjects were included. The patients were assessed using the Structured Clinical Interview for DSM-5 Disorders—Clinician Version during their 1–4-day-long hospitalization and were divided into two groups: with and without comorbid depression. Blood samples for the determination of NLRP3, interleukin-18 (IL-18), interleukin-1β (IL-1β), and hsCRP levels were analyzed using ELISA. NLRP3, IL-1β, IL-18, and hsCRP levels were significantly higher in myocardial infarction patients compared to the healthy group (p = 0.02, p < 0.001, p < 0.001, and p < 0.001, respectively). No significant difference was found between the myocardial groups with and without depression. However, in the logistic regression analysis, the NLRP3 variable in myocardial infarction patients was found to have a significant contribution to the likelihood of depression (p = 0.015, OR = 1.72, and CI = 1.11–2.66). The likelihood of depression is associated with increasing NLRP3 levels in myocardial infarction patients. However, this potential role should be further explored in a larger sample. [ABSTRACT FROM AUTHOR]

Objective To investigate the regulation and mechanism of Herkinorin preconditioning on NOD-like receptor protein 3 (NLRP3) in brain tissue of transient middle cerebral artery occlusion (tMCAO) rats. Methods Rats were randomly divided into sham operation group (sham), model group (tMCAO) and Herkinorin group (Herkinorin). Before modeling, rats in Herkinorin group were intraperitoneally given Herkinorin (10 mg/kg) once a day for a week. The rats were evaluated by neurologic score, TTC staining, TUNEL staining 24 h after reperfusion. The expression levels of IκBα, p65, p-p65, pro-IL-1β, IL-1β, pro-caspase1, caspase1 p20 and NLRP3 were detected by Western blot. The binding level of IκBα to β-arrestin2 was detected by co-immunoprecipitation (Co-IP). Results Compared with sham group, rats in tMCAO group had higher behavioral score (P＜0.01), increased cerebral infarction percentage and increased cerebral edema rate (P＜0.01). The level of cell apoptosis was increased in ischemic penumbra (P＜0.01). The expression levels of IκBα and p65 were decreased (P＜0.01) and the expression levels of p-p65, IL-1β, caspase1 p20 and NLRP3 were increased in ischemic penumbra (P＜0.01). The binding level of IκBα to β-arrestin2 was decreased in ischemic penumbra (P＜0.01). Compared with tMCAO group, rats in Herkinorin group had lower behavioral score (P＜0.01), lower cerebral infarction percentage and lower cerebral edema rate (P＜0.01). The level of cell apoptosis was decreased in ischemic penumbra(P＜0.01). The expression of IκBα and p65 was increased (P＜0.01) and the expression of p-p65, IL-1β, caspase1 p20 and NLRP3 was decreased in ischemic penumbra (P＜0.01). The binding of IκBα to β-arrestin2 was increased in ischemic penumbra (P＜0.01). Conclusions Herkinorin may negatively regulate NF-κB/NLRP3 pathway to alleviate cerebral ischemia-reperfusion injury of tMCAO rats.

Sepsis-induced myocardial dysfunction (SIMD) is one of the serious health-affecting problems worldwide. At present, the mechanisms of SIMD are still not clearly elucidated. The NOD-like receptor protein 3 (NLRP3) inflammasome has been assumed to be involved in the pathophysiology of SIMD by regulating multiple biological processes. NLRP3 inflammasome and its related signaling pathways might affect the regulation of inflammation, autophagy, apoptosis, and pyroptosis in SIMD. A few molecular specific inhibitors of NLRP3 inflammasome (e.g., Melatonin, Ulinastatin, Irisin, Nifuroxazide, and Ginsenoside Rg1, etc.) have been developed, which showed a promising anti-inflammatory effect in a cellular or animal model of SIMD. These experimental findings indicated that NLRP3 inflammasome could be a promising therapeutic target for SIMD treatment. However, the clinical translation of NLRP3 inhibitors for treating SIMD still requires robust in vivo and preclinical trials.

To investigate the effect of propofol pretreatment on microglial pyroptosis in the ischemic penumbra of rats subjected to cerebral ischemia-reperfusion. 90 rats were randomly divided into Sham group, model group, nod-like receptor protein 3 inhibitor group, propofol low dose group and high dose group. Pathological changes in the hippocampal region of the ischemic penumbra of brain tissue were observed by hematoxylin and eosin staining; interleukin-1 beta and interleukin-18 levels were detected in the ischemic penumbra of cerebral tissue by enzyme-linked immunosorbent assay; immunofluorescence double staining was performed to detect the immunopositive co-expression of nod-like receptor protein 3, aspartate specific caspase 1 and ionized calcium adaptor protein 1 in the ischemic penumbra of cerebral tissue. Compared with the model group, the nod-like receptor protein 3 inhibitor group, propofol low dose group and propofol high dose group groups also had significantly lower neurological deficit scores, cerebral infarction volumes, and interleukin-1 beta and interleukin-18 levels, ionized calcium adaptor protein 1/nod-like receptor protein 3, ionized calcium adaptor protein 1/caspase 1 immunopositive co-expression, nod-like receptor protein 3, pro-caspase 1, P10, P20 and interleukin-1 beta protein expressions in the ischemic penumbra were significantly reduced (p<0.05), the pathological damage was improved to various degrees, and the number of nerve cells was increased. The neuroprotective effect of propofol pretreatment on cerebral ischemia-reperfusion injury may be related to the inhibition of microglial pyroptosis mediated by nod-like receptor protein 3 inflammasome. [ABSTRACT FROM AUTHOR]

The inflammasome is a multimeric protein complex located in the cytoplasm that is activated by many factors and subsequently promotes the release of proinflammatory factors such as interleukin (IL)-1β and IL-18, resulting in a series of inflammatory responses that ultimately lead to the occurrence of various diseases. The Nod-like receptor protein 3 (NLRP3) inflammasome is the most characteristic type and the most widely studied among many inflammasomes. Activation of the NLRP3 inflammasome is closely related to the occurrence of many diseases, such as Alzheimer's disease. At present, a large number of studies have focused on the mechanisms underlying the activation of the NLRP3 inflammasome. Plenty of articles have reported the activation of the NLRP3 inflammasome by various ions, such as K+ and Na+ reflux and Ca2+ influx. However, few articles have reviewed the effects of various ion channels on the activation of the NLRP3 inflammasome and the relationship between the diseases caused by these proteins. This article mainly summarizes the relationship between intracellular and extracellular ion activities and ion channels and the activation of the NLRP3 inflammasome. We also provide a general summary of the diseases of each system caused by NLRP3 activation. We hope that more research will provide options for the treatment of diseases driven by the NLRP3 inflammasome. [ABSTRACT FROM AUTHOR]

Context Globally, the morbidity and mortality of cardiovascular diseases remain high. Didymin, a flavonoid glycoside, has long been used as a dietary antioxidant.Objective To determine the role of didymin in myocardial infarction (MI), and its possible myocardial protective mechanism.Materials and methods C57/BL6 mice (aged 6–8 weeks, n = 40) were divided into five groups: sham group, ischaemia–reperfusion (I/R) group, I/R + didymin (1 mg/kg) group, I/R + didymin (2 mg/kg) group and I/R + didymin (4 mg/kg) group. Didymin was administered intragastrically daily before I/R for 5 consecutive days. H9C2 cells were divided into five groups: control group, H/R group, H/R + didymin (3 μM) group, H/R + didymin (10 μM) group and H/R + didymin (30 μM) group. H9C2 cells were treated with didymin for 24 h before hypoxia/reoxygenation (H/R).Results In vivo, didymin reduced the pathological damage and fibrosis of myocardial tissues, decreased the levels of lactate dehydrogenase, creatine kinase, connective tissue growth factor, collagen I and collagen III. Moreover, didymin reduced myocardial apoptosis, inhibited NLRP3, ASC and caspase-1 expression, and alleviated the inflammatory response. In vitro, didymin reduced MI, apoptosis, inflammation and the levels of NLRP3, ASC and caspase-1 in H9C2.Discussion and conclusions Didymin prevented the deterioration of MI by inhibiting NLRP3 inflammasome in vivo and in vitro, and may be a potential natural drug for the treatment of MI. Our study provides the scientific basis for further research of didymin.

Objective To investigate the regulation and mechanism of Herkinorin preconditioning on NOD-like receptor protein 3 (NLRP3) in brain tissue of transient middle cerebral artery occlusion (tMCAO) rats. MethodsRats were randomly divided into sham operation group (sham), model group (tMCAO) and Herkinorin group (Herkinorin). Before modeling, rats in Herkinorin group were intraperitoneally given Herkinorin (10 mg/kg) once a day for a week. The rats were evaluated by neurologic score, TTC staining, TUNEL staining 24 h after reperfusion. The expression levels of IκBα, p65, p-p65, pro-IL-1β, IL-1β, pro-caspase1, caspase1 p20 and NLRP3 were detected by Western blot. The binding level of IκBα to β-arrestin2 was detected by co-immunoprecipitation (Co-IP). Results Compared with sham group, rats in tMCAO group had higher behavioral score (P＜0.01), increased cerebral infarction percentage and increased cerebral edema rate (P＜0.01). The level of cell apoptosis was increased in ischemic penumbra (P＜0.01). The expression levels of IκBα and p65 were decreased (P＜0.01) and the expression levels of p-p65, IL-1β, caspase1 p20 and NLRP3 were increased in ischemic penumbra (P＜0.01). The binding level of IκBα to β-arrestin2 was decreased in ischemic penumbra (P＜0.01). Compared with tMCAO group, rats in Herkinorin group had lower behavioral score (P＜0.01), lower cerebral infarction percentage and lower cerebral edema rate (P＜0.01). The level of cell apoptosis was decreased in ischemic penumbra (P＜0.01). The expression of IκBα and p65 was increased (P＜0.01) and the expression of p-p65, IL-1β, caspase1 p20 and NLRP3 was decreased in ischemic penumbra (P＜0.01). The binding of IκBα to β-arrestin2 was increased in ischemic penumbra (P＜0.01). Conclusions Herkinorin may negatively regulate NF-κB/NLRP3 pathway to alleviate cerebral ischemia-reperfusion injury of tMCAO rats. [ABSTRACT FROM AUTHOR]

The present study identified a novel upstream long chain non-coding (lncRNA) NEAT1/miR-141-3p/HTRA1 axis that regulated the activation of NLR family pyrin domain containing 3 (NLRP3) inflammasome to modulate endometriosis (EM) development. Specifically, clinical data suggested that the expression of NLRP3 and apoptosis-associated speck-like protein containing CARD (ASC), the cleavage of caspase-1 and gasdermin D (GSDMD), and the production of inflammatory cytokines (interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and IL-18) were all significantly increased in the ectopic endometrium (EE) tissues, compared to the normal endometrium (NE) tissues. Then, through analyzing the datasets from GEO database (GSE2339, GSE58178, and GSE7305) using the GEO2R bioinformatics tools, we verified that HtrA Serine Peptidase 1 (HTRA1) was especially enriched in the EE tissues compared to the NE tissues. To further confirm the biological functions of HTRA1, HTRA1 was overexpressed or downregulated in primary human endometrial stromal cells (hESCs) isolated from NE tissues or EE tissues, respectively. The results showed that upregulation of HTRA1 activated NLRP3 inflammasome-mediated pyroptotic cell death and cellular inflammation in NE-derived hESCs, whereas silencing of HTRA1 played an opposite role in EE-derived hESCs. In addition, the lncRNA NEAT1/miR-141-3p axis was screened as the upstream regulator of HTRA1. Mechanistically, lncRNA NEAT1 sponged miR-141-3p to positively regulate HTRA1 in a competing endogenous RNA (ceRNA) mechanisms-dependent manner. The recovery experiments in hESCs from NE and EE tissues confirmed that lncRNA NEAT1 overexpression promoted NLRP3 inflammasome-mediated pyroptotic cell death through regulating the miR-141-3p/HTRA1 axis. Taken together, this study firstly uncovered the underlying mechanisms by which a novel lncRNA NEAT1/miR-141-3p/HTRA1-NLRP3 pathway contributed to the development of EM, which provided novel diagnostic and therapeutic biomarkers for this disease. [ABSTRACT FROM AUTHOR]

We aimed to evaluate the cerebroprotective eff ects of Yizhitongmai Granule and its decomposed recipes on vascular dementia (VD) rats via the Nod-like receptor protein 3 (NLRP3) infl ammasome pathway. Sixty rats were randomly divided into Sham, VD Model, Yizhitongmai Recipe, Bushen Recipe, Tongluo Recipe and positive control groups (n=10). From 12 d aft er operation, Yizhitongmai Recipe, Bushen Recipe and Tongluo Recipe groups were gavaged with corresponding drug liquid. Th e drugs were administered at 2 mL once a day for 28 consecutive days. Th e reactive oxygen species (ROS), superoxide dismutase (SOD), total antioxidant capacity (T-AOC) and lactate dehydrogenase (LDH) in hippocampal tissues were detected using biochemical methods. Tumor necrosis factor-α (TNF-α), interleukin-18 (IL-18) and IL-1β were detected by enzyme-linked immunosorbent assay. Western blotting was performed to detect the expression levels of neuronal growth-associated protein-43 (GAP43), synaptophysin (SYN), aquaporin 4 (AQP4), NLRP3 and Caspase-1. Compared with the Model group, the number of apoptotic cells, levels of ROS, LDH, TNF-α, IL-18, IL-1β, NLRP3 and Caspase-1 decreased, and the levels of SOD, T-AOC, GAP43, SYN and AQP4 increased in the Yizhitongmai Recipe, Bushen Recipe and Tongluo Recipe groups (P0.05). Compared with the Bushen Recipe and Tongluo Recipe groups, the Yizhitongmai Recipe group had fewer apoptotic cells, decreased levels of ROS, LDH, TNF-α, IL-18, IL-1β, NLRP3 and Caspase-1, and increased levels of SOD, T-AOC, GAP43, SYN and AQP4 (P

Significance: The thioredoxin system comprises thioredoxin (Trx), thioredoxin reductase (TrxR), and nicotinamide adenine dinucleotide phosphate, besides an endogenous Trx inhibitor, the thioredoxin-interacting protein (TXNIP). The Trx system plays critical roles in maintaining the redox homeostasis in the central nervous system (CNS), in which oxidative stress damage is prone to occurrence due to its high-energy demand. Recent Advances: Increasing studies have demonstrated that the expression or activity of Trx/TrxR is usually decreased and that TXNIP expression is increased in patients with CNS diseases, including neurodegenerative diseases, cerebral ischemia, traumatic brain injury, and depression, as well as in their cellular and animal models. The compromise of Trx/TrxR enhances the susceptibility of neurons to related pathological state. Increased TXNIP not only enhances the inhibition of Trx activity, but also activates the NOD-like receptor protein 3 inflammasome, resulting in neuroinflammation in the brain. Critical Issues: In this review, we highlight the sources of oxidative stress in the CNS. The expression and function of the Trx system are summarized in different CNS diseases. This review also mentions that some inducers of Trx show neuroprotection in CNS diseases. Future Directions: Accumulating evidence has demonstrated the important roles of the Trx system in CNS diseases, suggesting that the Trx system may be a promising therapeutic target for CNS diseases. Further study should aim to develop the most effective inducers of Trx and specific inhibitors of TXNIP and to apply them in the clinical trials for the treatment of CNS diseases. Antioxid. Redox Signal. 38, 425–441. [ABSTRACT FROM AUTHOR]

Objective: To investigate the protective effect and mechanism of polydatin (PD) against gouty nephropathy (GN) in mice. Methods: Twenty‐four mice were randomly divided into three groups: the control group (no treatment), the GN group (300 mg/kg hypoxanthine + 150 mg/kg potassium oxonate), and the GN + PD group (300 mg/kg hypoxanthine + 150 mg/kg potassium oxonate + 50 mg/kg PD). Histological changes in the kidneys and the levels of uric acid (UA), blood urea nitrogen (BUN), and serum creatinine (SCr) in the sera were measured. In addition, the expression of gasdermin D (GSDMD) protein in renal tubular epithelial cells, and the expression of NOD‐like receptor protein 3 (NLRP3), GSDMD, and caspase‐1 proteins in the kidney tissues were determined by immunohistochemistry, immunofluorescence, and Western blot. Results: In vitro, PD inhibited the expression of NLRP3, caspase‐1, and GSDMD and protected the renal tubular epithelial cells from pyroptosis. In vivo, PD treatment significantly ameliorated the pathological changes in kidney tissue, and reversed the decrease of serum UA and BUN in GN model mice. The expression of NLRP3, GSDMD, and caspase‐1 proteins was also decreased in the PD‐treated GN mice. Conclusion: The results suggest that PD has a protective effect on mice with GN, which may be related to the downregulation of NLRP3, GSDMD, and caspase‐1 proteins and the inhibition of renal tubular epithelial cells pyroptosis. [ABSTRACT FROM AUTHOR]

Background: Regulatory T (Treg) cells are a class of anti-inflammatory lymphocyte subpopulations with a potential protective effect against atherosclerosis, whereas T helper 17 (Th17) cells have been reported to possess proatherogenic activity. It was believed that disturbed circulating Treg/Th17 balance was associated with the onset and progression of atherosclerosis. This study is designed to probe the regulative action of serum Nod-like receptor protein 3 (NLRP3) on the Treg/Th17 balance in patients with atherosclerosis. Methods: Fifty-two patients with coronary atherosclerosis and stenosis degrees of more than 50% were assigned to the coronary artery disease (CAD) group, and an equal number of people without coronary atherosclerosis were assigned to the control group (assessed by coronary angiography). Peripheral blood mononuclear cells (PBMCs) from two group patients were extracted and cultivated. The calculation of the Treg/Th17 ratio and quantitative analysis of the Treg and Th17 cell frequencies were performed through flow cytometry. Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was executed for the quantitative mRNA detection of the fork head- winged helix transcription factor (Foxp3) and the retinoic acid-related orphan nuclear receptor C (RORC) in PBMCs. Enzyme-linked immunosorbent assays were applied to measure the serum level of NLRP3, interleukin (IL)-10, IL-1β, IL-17A, IL-23, and transforming growth factor (TGF)-β1. Additionally, the connection between serum Treg/Th17 ratio and NLRP3 levels was analyzed using the Pearson correlation coefficient. Results: The baseline parameters, including sex, age, or blood biochemical indices had no difference in both groups (p > 0.05). The CAD group showed higher Th17 cell frequency, lower Treg cell frequency, and a lower Treg/Th17 ratio when compared to the control (p < 0.05). Consistent with the variation in the T-cell subset ratio, in patients with atherosclerosis, the Th17- cell-related transcription factor RORC showed a markedly higher mRNA level (p < 0.05), conversely, the mRNA expression of the Treg cell-related transcription factor Foxp3 was notably reduced (p < 0.05). Similarly, the serum levels of NLRP3, IL-17A, IL-1, and IL-23 were significantly enhanced in CAD group but IL-10 and TGF-β1 were reduced (p < 0.05). Additionally, a negative correlation was found between NLRP3 and the Treg/Th17 ratio (r = --0.69, p < 0.001). Conclusions: Due to the potential impact on the serum Treg/Th17 ratio, NLRP3 may act as an aggravator in the onset and progression of atherosclerotic disease. [ABSTRACT FROM AUTHOR]

Circulating oxidized linoleic acid (LA) metabolites (OXLAMs) are increased in patients with nonalcoholic steatohepatitis (NASH) and their levels correlate with disease severity. However, the mechanisms by which OXLAMs contribute to NASH development are incompletely understood. We tested the hypothesis that LA or OXLAMs provided directly through the diet are involved in the development of hepatic injury. C57BL/6 mice were fed an isocaloric high-fat diet containing low LA, high LA, or OXLAMs for 8 weeks. The livers of OXLAM-fed mice showed lower triglyceride concentrations, but higher FA oxidation and lipid peroxidation in association with increased oxidative stress. OXLAM-induced mitochondrial dysfunction was associated with reduced Complex I protein and hepatic ATP levels, as well as increased mitochondrial biogenesis and cytoplasmic mitochondrial DNA. Oxidative stress increased thioredoxin-interacting protein (TXNIP) in the liver and stimulated the activation of mitochondrial apoptosis signal-regulating kinase 1 (ASK1) leading to apoptosis. We also found increased levels of NOD-like receptor protein 3 (NLRP3) inflammasome components and Caspase-1 activation in the livers of OXLAM-fed mice. In vitro, OXLAMs induced hepatocyte cell death, which was partly dependent on Caspase-1 activation. This study identified key mechanisms by which dietary OXLAMs contribute to NASH development, including mitochondrial dysfunction, hepatocyte cell death, and NLRP3 inflammasome activation.

Chronic kidney disease (CKD) is a major public health problem with a developing incidence and prevalence. As a consequence of the growing number of patients diagnosed with renal dysfunction leading to the development of CKD, it is particularly important to explain the mechanisms of its underlying causes. In our paper, we discuss the molecular mechanisms of the development and progression of CKD, focusing on oxidative stress, the role of the immune system, neutrophil gelatinase-associated lipocalin, and matrix metalloproteinases. Moreover, growing evidence shows the importance of the role of the gut–kidney axis in the maintenance of normal homeostasis and of the dysregulation of this axis in CKD. Further, we discuss the therapeutic potential and highlight the future research directions for the therapeutic targeting of CKD. However, additional investigation is crucial to improve our knowledge of CKD progression and, more importantly, accelerate basic research to improve our understanding of the mechanism of pathophysiology. [ABSTRACT FROM AUTHOR]

Myocardial ischemia-reperfusion injury (MIRI) results in the aggravation of myocardial injury caused by rapid recanalization of the ischemic myocardium. In the past few years, there is a growing interest in investigating the complex pathophysiological mechanism of MIRI for the identification of effective targets and drugs to alleviate MIRI. Currently, pyroptosis, a type of inflammatory programmed death, has received greater attention. It is involved in the MIRI development in combination with other mechanisms of MIRI, such as oxidative stress, calcium overload, necroptosis, and apoptosis, thereby forming an intertwined association between different pathways that affect MIRI by regulating common pathway molecules. This review describes the pyroptosis mechanism in MIRI and its relationship with other mechanisms, and also highlights non-coding RNAs and non-cardiomyocytes as regulators of cardiomyocyte pyroptosis by mediating associated pathways or proteins to participate in the initiation and development of MIRI. The research progress on novel small molecule drugs, clinical drugs, traditional Chinese medicine, etc. for regulating pyroptosis can play a crucial role in effective MIRI alleviation. When compared to research on other mature mechanisms, the research studies on pyroptosis in MIRI are inadequate. Although many related protective drugs have been identified, these drugs generally lack clinical applications. It is necessary to further explore and verify these drugs to expand their applications in clinical setting. Early inhibition of MIRI by targeted regulation of pyroptosis is a key concern that needs to be addressed in future studies. [ABSTRACT FROM AUTHOR]

Diabetes mellitus can cause a decrease in tear secretion and tear film composition, resulting in a series of pathological changes such as the stability of tear film, the increased risk of inflammation, peripheral neuropathy and impairment of meibomian glands, leading to dry eye. The imbalance of inflammatory mediators in diabetic patients and the injury of ocular surface are the main factors that aggravate dry eye. Nod-like receptor protein 3 inflammasome was related to the occurrence and development of various systemic diseases. Evaluate the difference between nod-like receptor protein 3 inflammasome and diabetic dry eye parameters in diabetic dry eye patients, analyze the trend of correlation and discuss its mechanism. All subjects received teardrop break time, tear Schirmer’s I test, fluorescein staining score and objective scatter index measurement. The level of nod-like receptor protein 3, interleukin-1 beta, interleukin-6, interleukin-10, interleukin-18 in tears meanwhile nod-like receptor protein 3, caspase-1, tumor necrosis factor alpha, interleukin-1β, interleukin-6, interleukin-10, interleukin-18, superoxide dismutase, monoamine oxidase type B in serum was measured by double antibody sandwich enzyme-linked immunosorbent assay. In addition, the promoter gene expression levels of nod-like receptor protein 3, caspase-1, toll-like receptor 4, tumor necrosis factor alpha were determined by real-time quantitative real time polymerase chain reaction. BUT in diabetic dry eye patients was shortened, tear Schirmer’s I test and fluorescein staining scores were reduced and the levels of nod-like receptor protein 3, interleukin1β, interleukin-6 and interleukin-18 in tear increased significantly while the change of interleukin-10 was nonstatistical significance. Nod-like receptor protein 3, caspase-1, tumor necrosis factor alpha, interleukin-1β, interleukin-18 in serum were increased significantly and superoxide dismutase, monoamine oxidase type B were decreased statistically, meanwhile, levels of interleukin-6, interleukin-10 has no statistical difference. The results of real-time quantitative real time polymerase chain reaction showed that levels of nod-like receptor protein 3, caspase-1, toll-like receptor 4, tumor necrosis factor alpha were statistically increased also. Pearson correlation analysis showed that there was a strong positive correlation between nod-like receptor protein 3 levels and diabetic dry eye. The level of nod-like receptor protein 3 in diabetic dry eye increased significantly. There was a positive correlation between the clinical parameters and nod-like receptor protein 3 levels in diabetic dry eye, suggesting that regulation of nod-like receptor protein 3 level may provide new ideas for preventing and treating diabetic dry eye. [ABSTRACT FROM AUTHOR]

Copyright of Kafkas Universitesi Veteriner Fakultesi Dergisi is the property of University of Kafkas, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

NLRP3 inflammasome activation is a central process in initiating gout flares. The unique conformational rearrangement of the P2X7 receptor (P2X7R) upon ATP binding is critical for the activation of the NLRP3 inflammasome. However, studies on allosteric modulation of P2X7R in gout treatment are limited. Here, we aimed to investigate the therapeutic implications of targeting P2X7R in gout by designing a P2X7R allosteric inhibitor and validating the inhibitory function on NLRP3 inflammasome activation. Through virtual screening, we identified Z1456467176 (N-{3-[(2-aminoethyl) sulfamoyl] phenyl}-2-methyl-3-[3- (trifluoromethyl) phenyl] propanamide hydrochloride) bound to the drugbinding pocket as a potential antagonist of P2X7R. In functional assays, ATPor BzATP-induced P2X7R function was assessed in vitro in HEK-293T cells overexpressing hP2X7R (dye uptake assay) and macrophages (IL-1β release assay). Z1456467176 exhibited a stable and significant P2X7R inhibitory effect. Importantly, in MSU crystal-induced gout, the presence and involvement of ATP were confirmed. Z1456467176 blocked ATP-induced activation of the NLRP3- caspase-1-IL-1β pathway and exerted promising effects in reducing gouty joint inflammation in rats. In addition, molecular docking and molecular dynamics simulation studies showed that the P27XR protein conformation was remodeled by Z1456467176 binding. Collectively, our results provide a potent P2X7R allosteric inhibitor that facilitates the remission of MSU crystalinduced gout inflammation by inhibiting NLRP3 inflammasome activation, suggesting that allosteric inhibition of P2X7R represents a new direction in gout treatment. [ABSTRACT FROM AUTHOR]

Objective To investigate the neuroprotective mechanism of nilotinib in epileptic rats and the effect of COX-2/NLRP3/IL-18 signaling pathway. Methods Forty-eight rat epilepsy models were successfully established by intraperitoneal injection of pentylenetetrazole (PTZ) and then were randomly divided into the model group, SC-58125 (COX-2 inhibitor) group, nilotinib group, and nilotinib+SC-58125 group, with 12 in each group; another 12 normal rats were taken as the control group. The rats in the SC-58125 group were given 10 mg/kg SC-58125 by gavage every day, the rats in the nilotinib group were given 25 mg/kg nilotinib by gavage every day, and the rats in the nilotinib + SC-58125 group were given 25 mg/kg nilotinib and 10 mg/kg SC-58125 by gavage every day, and the rats in the control group and model group were given the same amount of solvent by gavage, once a day, for 4 consecutive weeks. One hour after the last administration, except for the control group, rats in the other groups were injected with PTZ again to observe the seizures of the rats, and the final seizure level, seizure latency and duration were recorded. Then, 6 rats were randomly selected from each group, the brains were collected through decapitation, and the hippocampal tissues were isolated. HE staining was used to observe the histopathological changes of hippocampus, and Western blotting was used to detect the expression of COX-2/ NLRP3/IL-18 pathway-related proteins in hippocampus. Results The rats in the control group had no seizures; the rats in the model group showed obvious seizures after the last injection of PTZ, and neuronal damage in hippocampal CA1 area was seen. Compared with the model group, the final seizure score decreased and the seizure duration of the rats in the SC-58125 group and the nilotinib group was shorter, the seizure latency was significantly prolonged, and the neuronal damage in the hippocampal CA1 area was alleviated (all P<0. 05). Compared with the control group, the expression levels of COX-2, NLRP3, Cleaved Caspase-1, IL-1β, and IL-18 in the hippocampus of the model group increased (all P<0. 05). Compared with the model group, the expression levels of COX-2, NLRP3, Cleaved Caspase-1, IL-1β and IL-18 in the the SC-58125 group and nilotinib group decreased (all P<0. 05). The expression levels of COX-2, NLRP3, Cleaved Caspase-1, IL-1β and IL-18 in the nilotinib + SC-58125 group were lower than those in the SC-58125 and nilotinib groups (all P< 0. 05). Conclusion Nilotinib may reduce PTZ-induced seizures and hippocampal neuron damage in rats by inhibiting the COX-2/NLRP3/IL-18 signaling pathway. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Knee osteoarthritis (KOA) is a major cause of disability in elderly individuals. Dicoumarol is a coumarin-like compound derived from sweet clover [Melilotus officinalis (L.) Pall]. It has been suggested that dicoumarol exhibits various types of pharmacological activities, including anticoagulant, antitumor and antibacterial effects. Due to its various biological activities, dicoumarol has a potential protective effect against OA. Therefore, the present study aimed to assess the effects of dicoumarol on knee osteoarthritis. In the present study, dicoumarol was found to protect rat synoviocytes from lipopolysaccharide (LPS)-induced cell apoptosis. Western blot analysis showed that dicoumarol significantly reduced the protein expression levels of fibrosis-related markers and inflammatory cytokines (Tgfb, Timp, Col1a, Il1b and Il18). The inhibitory rates of these proteins were all >50% (P<0.01) compared with those in the LPS and ATP-induced group. Consistently, the mRNA expression levels of these markers and cytokines were decreased to normal levels by dicoumarol after the treatment of rat synovial fibroblasts with LPS and ATP. Mechanistic studies demonstrated that dicoumarol did not affect NF-κB signaling, but it did directly interact with NOD-like receptor protein 3 (NLRP3) to promote its protein degradation, which could be reversed by MG132, but not NH4Cl. The protein half-life of NLRP3 was accelerated from 26.1 to 4.3 h by dicoumarol. Subsequently, dicoumarol could alleviate KOA in vivo; knee joint diameter was decreased from 11.03 to 9.93 mm. Furthermore, the inflammation and fibrosis of the knee joints were inhibited in rats. In conclusion, the present findings demonstrated that dicoumarol could impede the progression of KOA by inhibiting NLRP3 activation, providing a potential treatment strategy for KOA. [ABSTRACT FROM AUTHOR]

Kidney ischemia-reperfusion injury (IRI) is the major cause of poor prognosis after kidney transplantation and partial nephrectomy. Besides, it is also a critical pathophysiological process of acute kidney injury. Consequently, the prevention and treatment of kidney IRI are of significance to improve clinical prognosis of recipients undergoing kidney transplantation. However, the mechanism underlying IRI is complex, and the exact mechanism remains elusive. Inflammation, as one of the main pathogenesis of IRI, plays a significant role in IRI-induced kidney injury. Nuclear factor (NF)-κB, as a rapid response transcription factor, has been proven to be involved in the regulation of inflammation during kidney IRI. Therefore, in this article, the structure of NF-κB, the activation pattern of NF-κB signaling pathway, the regulatory mechanisms of NF-κB upstream and downstream signaling pathways in kidney IRI were reviewed, and the role of NF-κB signaling pathway in kidney IRI was investigated, aiming to provide novel clinical ideas for the prevention and treatment of kidney IRI. [ABSTRACT FROM AUTHOR]

Pyroptosis is a highly specific type of inflammatory programmed cell death that is mediated by Gasdermine (GSDM). It is characterized by inflammasome activation, caspase activation, and cell membrane pore formation. Diabetic cardiomyopathy (DCM) is one of the leading diabetic complications and is a critical cause of fatalities in chronic diabetic patients, it is defined as a clinical condition of abnormal myocardial structure and performance in diabetic patients without other cardiac risk factors, such as hypertension, significant valvular disease, etc. There are no specific drugs in treating DCM despite decades of basic and clinical investigations. Although the relationship between DCM and pyroptosis is not well established yet, current studies provided the impetus for us to clarify the significance of pyroptosis in DCM. In this review, we summarize the recent literature addressing the role of pyroptosis and the inflammasome in the development of DCM and summary the potential use of approaches targeting this pathway which may be future anti-DCM strategies. [ABSTRACT FROM AUTHOR]

Pyroptosis is a highly specific type of inflammatory programmed cell death that is mediated by Gasdermine (GSDM). It is characterized by inflammasome activation, caspase activation, and cell membrane pore formation. Diabetic cardiomyopathy (DCM) is one of the leading diabetic complications and is a critical cause of fatalities in chronic diabetic patients, it is defined as a clinical condition of abnormal myocardial structure and performance in diabetic patients without other cardiac risk factors, such as hypertension, significant valvular disease, etc. There are no specific drugs in treating DCM despite decades of basic and clinical investigations. Although the relationship between DCM and pyroptosis is not well established yet, current studies provided the impetus for us to clarify the significance of pyroptosis in DCM. In this review, we summarize the recent literature addressing the role of pyroptosis and the inflammasome in the development of DCM and summary the potential use of approaches targeting this pathway which may be future anti-DCM strategies.

Background: Electroacupuncture (EA) treatment has been found to ameliorate clinical symptoms in patients with dry eye, but its mechanisms are still not entirely clear. Objective: To study the regulation of EA on ocular surface function and the corneal reactive oxygen species (ROS)/thioredoxin-interacting protein (TXNIP)/Nod-like receptor protein 3 (NLRP3) inflammatory signaling pathway in dry eye syndrome (DES) model rats. Methods: Male Sprague-Dawley (SD) rats were randomly divided into five groups: Normal, Model, Model + EA, Model + NAC (N-actetylcysteine) and Model + NS (normal saline). The DES model was developed by subcutaneous injection of scopolamine hydrobromide with exposure to an air draft in the latter four groups. After intervention, the Schirmer I test (SIT), tear film break-up time (BUT) and ROS content were measured, the histopathological changes of corneal tissues were observed, and the mRNA and protein expression levels of TXNIP, NLRP3, apoptosis-associated Speck-like protein containing CARD (ASC), caspase-1, interleukin (IL)-1β and IL-18 were detected. Results: Compared with the Model group, the SIT and BUT increased significantly in the Model + EA group after intervention (p < 0.05), and the corneal injury was improved. Corneal ROS content declined in both Model + EA and Model + NAC groups (p < 0.05), and mRNA expression of TXNIP, NLRP3, ASC and caspase-1 also decreased (p < 0.01). Corneal protein expression of TXNIP, NLRP3, IL-1β and IL-18 decreased significantly in the Model + EA group (p < 0.01). Conclusion: Inhibiting the ROS/TXNIP/NLRP3 signaling pathway may be the mechanism underlying the role of EA in improving corneal injury in DES model rats. [ABSTRACT FROM AUTHOR]

Low-grade inflammation is a key mediator of the pathogenesis of Osteoarthritis (OA). Pulsed electromagnetic field (PEMF) can improve the symptoms of OA and potentially acts as an anti-inflammatory. The aim of this study was to investigate the effect of the PEMF on OA and its relationship with the NLRP3/Caspase-1/GSDMD signaling pathway.18 Three-month-old Sprague-Dawley (SD) rats were randomly divided into three groups (n = 6 per group): 1) OA group, 2) OA+PEMF group (OA with PEMF exposure), 3) Control group (sham operation with placebo PEMF). Rats in the OA and OA+PEMF groups were subjected to bilateral anterior cruciate ligament transection and ovariectomy. PEMF scheme: Pulse waveform, 3.82 mT, 8 Hz, 40 min/day, 5 days a week, for 12 weeks. The expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, and MMP-13 were detected by qRT-PCR and Western blot. The pathological structures of OA were monitored with Safranin O/fast green staining and hematoxylin eosin staining. Our results showed that PEMF alleviated the degree of inflammation and degeneration of cartilage in rats with OA, based on the histopathological changes and decline of the expression of IL-1β and MMP-13. Moreover, the over-expression of NLRP3, Caspase-1, and GSDMD in the cartilage of the OA rats decreased after PEMF treatment. These results suggested that PEMF could be a highly promising noninvasive strategy to slow down the progression of OA and inhibition of the NLRP3/Caspase-1/GSDMD signaling pathway might be involved in the beneficial effect of PEMF. [ABSTRACT FROM AUTHOR]

Platelet plays an important role in the progression of atherosclerosis. Recently it has been reported that myocardial infarction (MI) triggers megakaryopoiesis and thrombopoiesis in the bone marrow and leads to increased circulating platelets, which might contribute to the aggravation of atherosclerosis. However, the underlying mechanisms remain unclear. Here, we analyzed post-MI bone marrow tissue and found that MI induced an upregulation of bone marrow NOD-like Receptor Protein 3 (NLRP3) and subsequent secretion of IL-1β, an essential stimulator of megakaryopoiesis. Targeting NLRP3 using a specific inhibitor MCC950 reduced bone marrow IL-1β expression. Using bone marrow whole-mount immunofluorescence staining combined with flow cytometry, we demonstrated that MCC950 reduced megakaryocyte cellularity and maturity, and effectively attenuated the excessive platelet production after MI. Importantly, mice subjected to MI treated with MCC950 showed a higher survival rate compared with the only MI group. Taken together, this study shows that bone marrow NLRP3-IL-1β signal regulates megakaryocyte development and platelet production after myocardial infarction. It provides a new hint that pharmacological inhibition of NLRP3 might become a potential therapeutic approach for controlling excessive thrombopoiesis after MI. • Myocardial infarction activates bone marrow NLRP3-IL-1β which regulates megakaryocyte development and platelet production. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect and mechanism of artesunate on renal ischemia-reperfusion injury (IRI) in rat. Methods Twenty-five SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), low-dose artesunate group (ART-L group), high-dose artesunate group (ART-H group) and NLRP3 inflammasome inhibitor group (INF39 group), with 5 rats in each group. The levels of serum creatinine (Scr), blood urea nitrogen (BUN) and pathological damage of renal tissue were analyzed. The levels of tumor necrosis factor (TNF)-a and interleukin (IL)-6 in serum were quantitatively measured. The expression level ofIL-113 in renal tissues was determined by immunohistochemical staining. The expression levels of pyroptosis-related proteins were detected by fluorescent staining and Western blot. Results Compared with the Sham group, the levels ofS cr and BUN were higher, the renal tissue injury was aggravated, the expression levels of TNF-a, IL-6 and IL-1 13 were higher, and the expression levels of kidney injury molecule (KIM-1), pyroptosis-related protein NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase-1), G asdermin D (GSDMD) and IL-113 proteins were higher in the IRI group. Compared with the IRI group, the levels ofS cr and BUN were decreased, the renal tissue injury was mitigated, the expression levels ofT NF-a, IL-6 and IL-113 were down-regulated, and the expression levels ofK IM-1, NLRP3, Caspase-1, GSDMD and IL-113 proteins were down-regulated in the ART-L, ART-H and INF39 groups. Conclusions Artesunate may inhibit pyroptosis induced by NLR P3 inflammasome, down-regulate the expression levels of pyroptosis -related proteins, reduce the release of inflammatory factors after renal IRI and alleviate renal IRI. [ABSTRACT FROM AUTHOR]

The NOD-like receptor family pyrin domain-containing (NLRP3) inflammasomes is centrally implicated in cisplatin (CP)-induced kidney injury. Autophagy is critical for inhibiting production of NLRP3 protein that effectively reduces the inflammatory response. Ginsenoside Rg3 (SY), an active component extracted from ginseng, is reported to protect against CP-induced nephrotoxicity. However, the mechanisms underlying renoprotection by SY have not been established to date. Our results indicate that SY attenuated CP-induced apoptosis and damage in vivo and in vitro, as evidenced by increased cell viability, decreased the proportion of late apoptotic cells, elevated mitochondrial membrane potential, and ameliorated histopathological damage of the kidney. SY ameliorated CP-induced human renal tubular (HK-2) cells and kidney injury through upregulation of LC3II/I and beclin-1, inhibition of p62, NLRP3, ASC, caspase-1, and interleukin-1β. However, blockade of autophagy by 3-methyladenine reversed the suppression of SY on NLRP3 inflammasome activation and the protection of SY on HK-2 cells. Our collective results support the utility of SY as a therapeutic agent that effectively protects against CP-induced kidney injury by activating the autophagy-mediated NLRP3 inhibition pathway. [ABSTRACT FROM AUTHOR]

S-adenosylhomocysteine (SAH) is hydrolyzed by SAH hydrolase (SAHH) to homocysteine and adenosine. Increased plasma SAH levels were associated with disturbed renal function in patients with diabetes. However, the role and mechanism of SAHH in diabetic nephropathy is still unknown. In the present study, we found that inhibition of SAHH by using its inhibitor adenosine dialdehyde (ADA) accumulates intracellular or plasma SAH levels and increases high glucose-induced podocyte injury and aggravates STZ-induced diabetic nephropathy, which is associated with Nod-like receptor protein 3 (NLRP3) inflammasome activation. Inhibition or knockout of NLRP3 attenuates SAHH inhibition-aggravated podocyte injury and diabetic nephropathy. Additionally, SAHH inhibition increases thioredoxin-interacting protein (TXNIP)-mediated oxidative stress and NLRP3 inflammasome activation, but these effects were not observed in TXNIP knockout mice. Mechanistically, SAHH inhibition increased TXNIP by inhibiting histone methyltransferase enhancer of zeste homolog 2 (EZH2) and reduced trimethylation of histone H3 lysine 27 and its enrichment at promoter of early growth response 1 (EGR1). Moreover, EGR1 is activated and enriched at promoters of TXNIP by SAHH inhibition and is essential for SAHH inhibition-induced TXNIP expression. Inhibition of EGR1 protected against SAHH inhibition-induced NLRP3 inflammasome activation and oxidative stress and diabetic nephropathy. Finally, the harmful effects of SAHH inhibition on inflammation and oxidative stress and diabetic nephropathy were also observed in heterozygote SAHH knockout mice. These findings suggest that EZH2/EGR1/TXNIP/NLRP3 signaling cascade contributes to SAHH inhibition-aggravated diabetic nephropathy. Our study firstly provides a novel insight into the role and mechanism of SAHH inhibition in diabetic nephropathy.

Chronic kidney disease (CKD) is a major public health problem with a developing incidence and prevalence. As a consequence of the growing number of patients diagnosed with renal dysfunction leading to the development of CKD, it is particularly important to explain the mechanisms of its underlying causes. In our paper, we discuss the molecular mechanisms of the development and progression of CKD, focusing on oxidative stress, the role of the immune system, neutrophil gelatinase-associated lipocalin, and matrix metalloproteinases. Moreover, growing evidence shows the importance of the role of the gut–kidney axis in the maintenance of normal homeostasis and of the dysregulation of this axis in CKD. Further, we discuss the therapeutic potential and highlight the future research directions for the therapeutic targeting of CKD. However, additional investigation is crucial to improve our knowledge of CKD progression and, more importantly, accelerate basic research to improve our understanding of the mechanism of pathophysiology.

Myocardial ischemia-reperfusion injury (MIRI) results in the aggravation of myocardial injury caused by rapid recanalization of the ischemic myocardium. In the past few years, there is a growing interest in investigating the complex pathophysiological mechanism of MIRI for the identification of effective targets and drugs to alleviate MIRI. Currently, pyroptosis, a type of inflammatory programmed death, has received greater attention. It is involved in the MIRI development in combination with other mechanisms of MIRI, such as oxidative stress, calcium overload, necroptosis, and apoptosis, thereby forming an intertwined association between different pathways that affect MIRI by regulating common pathway molecules. This review describes the pyroptosis mechanism in MIRI and its relationship with other mechanisms, and also highlights non-coding RNAs and non-cardiomyocytes as regulators of cardiomyocyte pyroptosis by mediating associated pathways or proteins to participate in the initiation and development of MIRI. The research progress on novel small molecule drugs, clinical drugs, traditional Chinese medicine, etc. for regulating pyroptosis can play a crucial role in effective MIRI alleviation. When compared to research on other mature mechanisms, the research studies on pyroptosis in MIRI are inadequate. Although many related protective drugs have been identified, these drugs generally lack clinical applications. It is necessary to further explore and verify these drugs to expand their applications in clinical setting. Early inhibition of MIRI by targeted regulation of pyroptosis is a key concern that needs to be addressed in future studies.

Rupture of atherosclerotic plaques constitutes the major cause of thrombosis and acute ischemic coronary syndrome. Bone marrow‐derived mesenchymal stem cells microvesicles (BMSCs‐MVs) are reported to promote angiogenesis. This study investigated the role of BMSCs‐MVs in stabilizing atherosclerotic plaques. BMSCs‐MVs in mice were isolated and identified. The mouse model of atherosclerosis was established, and mice were injected with BMSCs‐MVs via the tail vein. The macrophage model with high glucose and oxidative damage was established and then incubated with BMSCs‐MVs. Nod‐like receptor protein 3 (NLRP3) expression, pyroptosis‐related proteins, and inflammatory factors were detected. Actinomycin D was used to inhibit the secretion of BMSCs‐MVs to verify the source of microRNA‐223 (miR‐223). The binding relationship between miR‐223 and NLRP3 was predicted and verified. BMSCs‐MVs with knockdown of miR‐223 were cocultured with bone marrow‐derived macrophages with knockdown of NLRP3, and then levels of miR‐223, NLRP3, pyroptosis‐related proteins, and inflammatory factors were detected. BMSCs‐MVs could reduce the vulnerability index of atherosclerotic plaques and intima‐media thickness in mice, and inhibit pyroptosis and inflammation. BMSCs‐MVs inhibited pyroptosis and inflammatory factors in macrophages. BMSCs‐MVs carried miR‐223 to inhibit NLRP3 expression and reduce macrophage pyroptosis, thereby stabilizing the atherosclerotic plaques. [ABSTRACT FROM AUTHOR]

Endothelial cell dysfunction is the main pathology of atherosclerosis (AS). Sirtuin 6 (SIRT6), a deacetylase, is involved in AS progression. This study aimed to investigate the impacts of SIRT6 on the pyroptosis of endothelial cells and its underlying mechanisms. ApoE-/- mice were fed a high-fat diet (HFD) to establish the AS mouse model, atherosclerotic lesions were evaluated using oil red O staining, and blood lipids and inflammatory factors were measured using corresponding kits. Human umbilical vein endothelial cells (HUVECs) were treated with oxidized low-density lipoprotein (ox-LDL) to establish the cell model, and pyroptosis was evaluated by flow cytometry, ELISA, and western blot. Immunoprecipitation (IP), co-IP, western blot, and immunofluorescence were used to detect the molecular mechanisms. The results showed that SIRT6 expression was downregulated in the blood of HFD-induced mice and ox-LDL-induced HUVECs. Overexpression of SIRT6 reduced atherosclerotic lesions, blood lipids, and inflammation in vivo and suppressed pyroptosis of HUVECs in vitro. Moreover, SIRT6 interacted with ASC to inhibit the acetylation of ASC, thus, reducing the interaction between ASC and NLRP3. Moreover, SIRT6 inhibits endothelial cell pyroptosis in the aortic roots of mice by deacetylating ASC. In conclusion, SIRT6 deacetylated ASC to inhibit its interaction with NLRP3 and then suppressed pyroptosis of endothelial cells, thus, decelerating the progression of AS. The findings provide new insights into the function of SIRT6 in AS.

Introduction: NOD-like receptor protein 3 (NLRP3) is implicated in chronic obstructive pulmonary disease (COPD) pathogenesis. Here, we explored the role of NLRP3 in cigarette smoke (CS)-induced airway inflammation in COPD., Material and Methods: NLRP3 expression level was assessed with the microarray data in GEO datasets and validated in serum by ELISA from a case-control cohort. Male C57BL/6J mice were randomly divided into: saline, CS, MCC950 (a specific NLRP3 inhibitor, 10 mg/kg) and CS + MCC950 (5 mg/kg and 10 mg/kg) groups ( n = 5 per group). All mice were exposed to CS or air for 4 weeks. Then, broncho-alveolar lavage (BAL) fluid and lung tissues were collected for cell counting, ELISA, HE staining and RNA sequencing with validation by real-time qPCR., Results: Compared to non-smokers, NLRP3 expression was significantly elevated in the lung tissues and sera of COPD smokers. CS remarkably induced airway inflammation in mice, characterized by an increase of inflammatory cells and proinflammatory cytokines in BAL fluid and HE inflammatory score, which were ameliorated by MCC950 treatment dose-dependently. Subsequently, 84 candidate genes were selected following RNA sequencing, and five hub genes (Mmp9, IL-1α, Cxcr2, Cxcl10, Ccr1) were then identified by PPI and MCODE analyses, which were confirmed by real-time qPCR. GO and KEGG analysis suggested that the five genes were enriched in a complicated network of inflammatory processes and signaling pathways., Conclusions: NLRP3 expression is elevated in lungs and sera of COPD smokers. Inhibition of NLRP3 significantly attenuates CS-induced airway inflammation in mice via inactivation of multiple hub genes and their related inflammatory processes and signaling pathways., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2024 Termedia & Banach.)

Abstract Inflammasomes are large intracellular multi-protein signalling complexes that are formed in the cytosolic compartment as an inflammatory immune response to endogenous danger signals. The formation of the inflammasome enables activation of an inflammatory protease caspase-1, pyroptosis initiation with the subsequent cleaving of the pro-inflammatory cytokines interleukin (IL)-1β and proIL-18 to produce active forms. The inflammasome complex consists of a Nod-like receptor (NLR), the adapter apoptosis-associated speck-like (ASC) protein, and Caspase-1. Dysregulation of NLRP3 inflammasome activation is involved tumor pathogenesis, although its role in cancer development and progression remains controversial due to the inconsistent findings described. In this review, we summarize the current knowledge on the contribution of the NLRP3 inflammasome on potential cancer promotion and therapy.

Circulating oxidized linoleic acid (LA) metabolites (OXLAMs) are increased in patients with nonalcoholic steatohepatitis (NASH) and their levels correlate with disease severity. However, the mechanisms by which OXLAMs contribute to NASH development are incompletely understood. We tested the hypothesis that LA or OXLAMs provided directly through the diet are involved in the development of hepatic injury. C57BL/6 mice were fed an isocaloric high-fat diet containing low LA, high LA, or OXLAMs for 8 weeks. The livers of OXLAM-fed mice showed lower triglyceride concentrations, but higher FA oxidation and lipid peroxidation in association with increased oxidative stress. OXLAM-induced mitochondrial dysfunction was associated with reduced Complex I protein and hepatic ATP levels, as well as increased mitochondrial biogenesis and cytoplasmic mitochondrial DNA. Oxidative stress increased thioredoxin-interacting protein (TXNIP) in the liver and stimulated the activation of mitochondrial apoptosis signal-regulating kinase 1 (ASK1) leading to apoptosis. We also found increased levels of NOD-like receptor protein 3 (NLRP3) inflammasome components and Caspase-1 activation in the livers of OXLAM-fed mice. In vitro, OXLAMs induced hepatocyte cell death, which was partly dependent on Caspase-1 activation. This study identified key mechanisms by which dietary OXLAMs contribute to NASH development, including mitochondrial dysfunction, hepatocyte cell death, and NLRP3 inflammasome activation.

Acute lung injury (ALI) is a common critical illness in clinical anesthesia and the intensive care unit that can cause acute hypoxic respiratory insufficiency. Despite various therapeutic regimes having been investigated, there is currently no effective pharmacotherapy available to treat ALI. Previous studies have reported that the NOD-like receptor protein 3 (NLRP3) signaling pathway plays an important role in the inflammatory response and is involved in the pathogenesis of ALI. Moreover, dexmedetomidine (Dex), an α2-adrenergic receptor activating agent, has been routinely used as an adjuvant therapy in treating inflammatory diseases, including ALI. However, the precise pathological mechanisms of Dex in ALI remain to be elucidated. Thus, the present study aimed to investigate the effects of the p38/heme oxygenase 1 (HO-1) signaling pathways in the pathological mechanisms of Dex in ALI. Newborn male Sprague-Dawley rats (n=48) were randomly divided into four groups (n=12 each), and an intravenous injection of lipopolysaccharide (LPS) was used to successfully induce the ALI model, with increased pulmonary damage, cell apoptosis, interleukin-1β (IL-1β) secretion and edema fluid in lungs. Moreover, the mRNA and protein expression levels of NLRP3 were significantly upregulated, while that of HO-1 were downregulated by LPS treatment. Furthermore, the levels of phosphorylated p38 were also upregulated in ALI rats. It was demonstrated that Dex administration significantly alleviated LPS-induced ALI, downregulated the secretion of IL-1β, decreased the expression of NLRP3, inhibited the phospho-activation of p38 and increased HO-1 expression. In addition, pharmacological inhibition of p38 using the inhibitor SB20380 further enhanced the effect of Dex. Collectively, these preliminarily results identified the effects of Dex intervention on the pathogenesis of ALI via the regulation of p38/HO-1 signaling pathways, which impacted the inflammatory effects, thus providing a theoretical basis and novel evidence for the development of new targets for clinical treatment of ALI. [ABSTRACT FROM AUTHOR]

Expression levels of miR-223-3p and NLRP3 in high glucose and high fat (HGHF)-induced diabetic mice, and the mechanism on the injury of mouse cardiac microvascular endothelial cells (MCMECs) were investigated. Four-week C57BL/6J laboratory mice were selected and randomized into a control group and a model group (n=10 each). Mice in the model group were fed with HGHF diet to establish a mouse model of diabetes. Further MCMECs were purchased to construct carriers through transient transfection, and were separated into a normal group (cultured in the normal environment), a model group (not transfected), a blank carrier group (transfected with miR-NC), a miR-223-3p-mimics group, and a miR-223-3p-inhibitor group. RT-qPCR was used to detect the expression levels of miR-223-3p and NLRP3, and western blot analysis to detect the expression levels of NLRP3, apoptosis-related proteins Bax and caspase-3, and anti-apoptotic protein Bcl-2. Flow cytometry was used to observe apoptosis and TargetScan to predict the target relationship between miR-223-3p and NLRP3. Dual-luciferase reporter gene assay was used to detect the relationship between miR-223-3p and NLRP3. Compared with those in the control group, the mice in the model group had significantly lower expression of miR-223-3p. However, significantly higher mRNA and protein expression levels of NLRP3 were observed (P<0.05). After modeling, miR-223-3p overexpression downregulated the expression levels of NLRP3 mRNA, Bax and NLRP3 protein, as well as inhibited endothelial cell apoptosis (P<0.05), while the inhibition of miR-223-3p expression upregulated the expression levels and promoted apoptosis. In conclusion, miR-223-3p expression is low, however, NLRP3 is highly expressed in the heart tissue of HGHF-induced diabetic mice. miR-223-3p reduces the injury of MCMECs and inhibits endothelial cell apoptosis in mice by regulating the expression of NLRP3. [ABSTRACT FROM AUTHOR]

Selenium (Se) is important in many physiological processes, such as antioxidant processes and inflammation. The aim of our experiments was to investigate the molecular mechanism that selenomethionine could reduce the lipopolysaccharide (LPS)-induced inflammation by inhibiting the TLR4-NF-κB-NLRP3 signaling pathway. Eighty broilers were randomly and evenly divided into two groups, giving normal Se content diets (Con group, 0.2 mg Se/kg diet) and Se-rich basal diets (Se group, 0.5 mg selenomethionine/kg diet) for 90 days. Se-rich basal diets were based on 0.2 mg/kg sodium selenite contained. Five hours before euthanized, 20 broilers were randomly selected from each group and given lipopolysaccharide (200 μg/kg BW) by intraperitoneal injection, Con+LPS group and Se+LPS group, respectively. The Con group and Se group were given equal saline by intraperitoneal injection. We observed the microscopic pathological changes of liver tissue detected oxidative stress by kit and detected the expression of inflammatory factors, heat shock protein (HSP), and nod-like receptor protein 3 (NLRP3)–related genes by qRT-PCR and Western blot. With the microscope, we found the Con+LPS group had obvious inflammatory lesions such as sinusoidal congestion, but the damage was significantly alleviated in the Se+LPS group. In the Con+LPS group, the activity of GSH-Px and the content of GSH were significantly decreased compared with those in the Con group; however, they are increased in the Se group and in the Se + LPS group. Inflammatory factors (MyD88, NF-κB, TNF-α, IL-1β, IL-6, IL-12, IL-18, iNOS, and COX-2), heat shock proteins (HSP27, HSP60, HSP70, and HSP90), and the expression of NLRP3 and caspase-1 increased in the Con+LPS group compared with those in the Con group, while they were lower in the Se+LPS group than in the Con+LPS group. We concluded that selenomethionine inhibits the LPS-induced inflammation of liver tissue via suppressing the TLR4-NF-κB-NLRP3 signaling pathway. [ABSTRACT FROM AUTHOR]

The limitation of doxorubicin (DOX), which is widely used for the treatment of solid tumors and hematologic malignancies, is a vital problem in clinical application. The most serious of limit factors is cardiotoxicity. Calycosin (CA), an isoflavonoid that is the major active component in Radix astragali, has been reported in many bioactivities including antitumor, anti-inflammatory, and cardioprotection. The aim of the study was to investigate the effects and mechanisms of CA on DOX-induced cardiotoxicity in vitro and in vivo. CA increased H9c2 cell viability and reduced apoptosis induced by DOX via Bcl-2, Bax, and the PI3K-Akt signaling pathway. Moreover, CA prevented DOX-induced oxidative stress in cells by decreasing the generation of reactive oxygen species. Similarly, oxidative stress was inhibited by CA through the increased activities of antioxidant enzymes such as glutathione peroxidase, catalase, and superoxide dismutase and decreased the levels of aspartate aminotransferase, lactate dehydrogenase, and malondialdehyde in vivo. Furthermore, the levels of sirtuin 1 (Sirt1)-NOD-like receptor protein 3 (NLRP3) and related proteins were ameliorated by CA in cells and in mice hearts. When H9c2 cells were treated by Ex527 (Sirt1 inhibitor), the effect of CA on expressions of NLRP3 and thioredoxin-interacting protein was suppressed. In conclusion, the results suggested that CA might be a cotreatment with DOX to ameliorate cardiotoxicity by Sirt1-NLRP3 pathway. [ABSTRACT FROM AUTHOR]

• NP induced the decline of spatial learning and memory and motor ability in rats. • NP led to the down-regulation of miR-542-3p and pyroptosis in substantia nigra. • Up-regulation of miR-542-3p targetd TLR4 and inhibit NP-induced microglial pyroptosis. This study aimed to investigate the spatial learning/memory and motor abilities of rats and the alteration of miR-542-3p and pyroptosis in the midbrain nigrostriatal area in vivo after nonylphenol (NP) gavage and to explore the mechanism of miR-542-3p regulation of Toll-like receptor 4 (TLR4) in NP-induced pyroptosis in BV2 microglia in vitro. In vivo : Thirty-six specific-pathogen-free-grade Sprague–Dawley rats were divided into three equal groups: blank control group (treated with pure corn oil), NP group (treated with NP, 80 mg/kg body weight per day for 90 days), and positive control group [treated with lipopolysaccharide (LPS), 2 mg/kg body weight for 7 days]. In vitro : The first part of the experiment was divided into blank group (control, saline), LPS group [1 µg/ml + 1 mM adenosine triphosphate (ATP)], and NP group (40 µmol/L). The second part was divided into mimics NC (negative control) group, miR-542-3p mimics group, mimics NC + NP group, and miR-542-3p mimics + NP group. In vivo : Behaviorally, the spatial learning/memory and motor abilities of rats after NP exposure declined, as detected via Y-maze, open field, and rotarod tests. Some microglia in the substantia nigra of the NP-treated rats were activated. The downregulation of miR-542-3p was observed in rat brain tissue after NP exposure. The mRNA/protein expression of pyroptosis-related indicators (TLR4), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), gasdermin-D (GSDMD), cysteinyl aspartate–specific proteinase-1 (caspase-1), and interleukin-1β (IL-1β) in the substantia nigra of the midbrain increased after NP exposure. In vitro : ASC fluorescence intensity increased in BV2 cells after NP exposure. The mRNA and/or protein expression of pyroptosis-related indicators (TLR4, NLRP3, GSDMD, caspase-1, and IL-1β) in BV2 cells was upregulated after NP exposure. The transfection of miR-542-3p mimics inhibited NP-induced ASC expression in BV2 cells. The overexpression of miR-542-3p, followed by NP exposure, significantly reduced TLR4, NLRP3, ASC, caspase-1, and IL-1β gene and/or protein expression. This study suggested that NP exposure caused a decline in spatial learning memory and whole-body motor ability in rats. Our study was novel in reporting that the upregulation of miR-542-3p targeting and regulating TLR4 could inhibit NLRP3 inflammatory activation and alleviate NP-induced microglia pyroptosis. [Display omitted] [ABSTRACT FROM AUTHOR]

Ageratina adenophora is an invasive weed with potent toxicological effects on livestock. Oxidative stress and pyroptosis play a pivotal role in regulating animal or human health and disease. The object of this study was to determine the mechanism underlying splenic toxicity induced by A. adenophora in a mouse model. Ageratina adenophora significantly increased the levels of reactive oxygen species and malondialdehyde, but decreased the antioxidants like catalase, superoxide dismutase, glutathione and glutathione peroxidase. In addition, the activity of the antioxidant enzymes was also decreased upon A. adenophora treatment. The induction of the pyroptosis pathway was evaluated in terms of the expression levels of Nod-like receptor protein 3, nuclear factor-κB, caspase-1, gasdermin-D and interleukin-1β, all of which were significantly elevated by A. adenophora. These findings suggest that A. adenophora impairs spleen function in mice through oxidative stress damage and pyroptosis.