Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 59
Filtrar
1.
Life Sci Alliance ; 6(2)2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36450446

RESUMO

Whereas the role of the nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing protein (NLRP) 3 pathway in innate immunity has been extensively studied, little attention has been paid to its contribution to adaptive immunity. Studies in animal models and human subjects have shown the contribution of NLRP3 to the T cell compartment, and its role in B lymphocyte functions has been proposed. Here, we report that ablation of nlrp3 in mice led to altered B cell development in the bone marrow, and distorted expression of B cell subsets that play innate-like functions, that is, marginal zone B cells in the spleen and B-1a cells in the peritoneal cavity. Mechanistically, in the absence of NLRP3 expression, the transcription factor IRF4, previously found to interact with NLRP3 in the nucleus of lymphocytes, was up-regulated. NLRP3 ablation reduced the expression of the chemokine receptors CXCR4 and CCR7 in an IRF4-dependent manner, indicating that the presence of NLRP3 is critical for optimal expression of chemokine receptors on B cells. We conclude that activation of the NLRP3 inflammasome plays a role in B cell development, homing, and retention in lymphoid organs.


Assuntos
Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Humanos , Animais , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Ativação Linfocitária , Domínio Pirina , Receptores de Quimiocinas
2.
Pharm Biol ; 60(1): 2319-2327, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36416076

RESUMO

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.


Assuntos
Infarto do Miocárdio , Traumatismo por Reperfusão Miocárdica , Camundongos , Animais , Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Traumatismo por Reperfusão Miocárdica/metabolismo , Domínio Pirina , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Glicosídeos , Infarto do Miocárdio/metabolismo , Caspase 1/metabolismo
3.
J Tradit Chin Med ; 42(6): 965-971, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36378055

RESUMO

OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome (CPPS) rats by electroacupuncture (EA) of Guanyuan (CV4), Zhongji (CV3), Huiyang (BL35) and Sanyinjiao (SP6), and to explore the possible mechanism of anti-inflammatory and analgesic effects of EA. METHODS:A total of 36 Sprague-Dawley male rats were randomly divided into three groups: control, model and EA (n=12 rats/group). The CPPS model was made by injection of CFA into ventral lobes of the prostate (0.1 mL). Electric acupuncture apparatus was applied to stimulate Guanyuan (CV4), Zhongji (CV3), bilateral Huiyang (BL35) and Sanyinjiao (SP6) acupoints in EA group. The general condition of rats was observed and the prostate index (PI) was calculated. The thermal pain threshold was collected after each therapeutic course. Histopathological changes of the prostate tissue were examined by hematoxylin-eosin staining method. The expression levels of tumor necrosis factor α (TNF-α), interleukin-1ß (IL-1ß) and prostaglandin E2 (PGE2) in prostatic homogenates were measured by enzyme linked immunosorbent assay (ELISA). Moreover, the expression levels of purinergic 2X7 receptor (P2X7R), NOD-like receptor pyrin domain-containing 3 (NLRP3), caspase-1 and interleukin-18 (IL-18) mRNA were quantified by quantitative real-time polymerase chain reaction. RESULTS: Compared with control group, the PI of rats increased, and the thermal pain threshold decreased significantly in model group. The morphological structure of prostate tissues of rats in model group was severely damaged with a large number of inflammatory cells infiltration. Additionally, the levels of TNF-α, IL-1ß and PGE2 were higher, and the expressions of P2X7R, NLRP3, caspase-1 and IL-18 mRNA were higher than those in control group. After EA treatment, the PI was significantly decreased, the thermal pain threshold was significantly increased, and the tissue damage was significantly improved. The expressions of inflammatory cytokines were lower in EA group, and expression of P2X7R/NLRP3 pathway was down-regulated. CONCLUSION: The effect of EA at Guanyuan (CV4), Zhongji (CV3), Huiyang (BL35) and Sanyinjiao (SP6) can improve inflammation and pain symptoms of CPPS rats induced by Complete Freund's adjuvant (CFA). This suggests that EA at Guanyuan (CV4), Zhongji (CV3), Huiyang (BL35) and Sanyinjiao (SP6) can produce anti-inflammatory analgesia effect by preventing the activation of P2X7R/NLRP3 signal pathway, inhibit the release of inflammatory cytokines in CPPS rats, which may provide a putative novel target for the treatment of CPPS.


Assuntos
Dor Crônica , Eletroacupuntura , Ratos , Masculino , Animais , Interleucina-18 , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Domínio Pirina , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/genética , Dinoprostona , Dor Pélvica/genética , Dor Pélvica/terapia , Adjuvante de Freund , Transdução de Sinais , Citocinas , RNA Mensageiro , Caspases
4.
Sci Rep ; 12(1): 19528, 2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36376416

RESUMO

NOD-like receptor pyrin domain containing 3 (NLRP3) is a microbial and danger signal sensor that acts as a regulator of inflammation via activation of Caspase-1 (CASP1) and has been identified as a major contributor to human liver diseases. The present study was conducted to investigate the association between NLRP3 and the progression of hepatitis C virus (HCV)-related liver disease. Serum NLRP3 levels were analyzed in 49 patients with chronic HCV infection and 18 healthy controls and liver tissues from 34 patients were examined to assess the protein expression of NLRP3 and its activation marker CASP1 using immunohistochemical staining. The results showed that the median serum NLRP3 levels was significantly higher in HCV-infected patients compared with healthy controls (1040 pg/ml vs 695 pg/ml respectively, P < 0.001) and were positively correlated with hepatic NLRP3 and CASP1 expression (r = 0.749, P < 0.001 and r = 0.557, P = 0.001 respectively). The NLRP3 levels in serum and the liver significantly increased with worsening liver pathology and showed positive correlations with serum aminotransferases levels, HCV viremia, and albumin-bilirubin score (P < 0.05). The receiver operating characteristic curve analysis revealed a high diagnostic performance of serum NLRP3 in determining the extent of liver necroinflammation, fibrosis, and steatosis (area under the curve = 0.951, 0.971, and 0.917 respectively, P < 0.001). In conclusion, NLRP3 plays an important role in liver disease progression during HCV infection via CASP1 activation and might be a promising therapeutic target. Serum NLRP3 could be an additional biomarker for liver inflammation and fibrosis.


Assuntos
Hepacivirus , Hepatite C Crônica , Humanos , Caspase 1/metabolismo , Fibrose , Hepacivirus/metabolismo , Inflamassomos/metabolismo , Fígado/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteínas NLR/metabolismo , Domínio Pirina
5.
Int J Mol Sci ; 23(21)2022 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-36361886

RESUMO

The nucleotide-binding domain (NOD)-, leucine-rich repeat (LRR)-, and pyrin domain (PYD)-containing protein 3, NLRP3, is a multiprotein complex belonging to the innate immune system that can be activated by pathogens or danger-associated molecular patterns [...].


Assuntos
Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Domínio Pirina
6.
Chem Biol Interact ; 365: 110122, 2022 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-36002070

RESUMO

Psoriasis is a common chronic autoinflammatory/autoimmune skin disease associated with elevated pro-inflammatory cytokines. The pivotal role of interleukin (IL)-1ß and nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome in the pathogenesis of psoriasis has been widely described. Accordingly, the suppression of NLRP3-dependent IL-1ß release is a potential therapy for psoriasis. Repurposing marketed drugs is a strategy for identifying new inhibitors of NLRP3 inflammasome activation. Herein, chlorquinaldol (CQD), a historic antimicrobial agent used as a topical treatment for skin and vaginal infections, was found to have a distinct effect by inhibiting NLRP3 inflammasome activation at concentrations ranging from 2 to 6 µM. CQD significantly suppressed apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) oligomerization, NLRP3-ASC interaction, and pyroptosis in macrophages. The levels of cleaved IL-1ß and caspase-1 were reduced by CQD in the cell lysates of macrophages, suggesting that CQD acted on upstream of pore formation in the cell membrane. Mechanistically, CQD reduced mitochondrial reactive oxygen species production but did not affect the nuclear factor-κB (NF-κB) pathway. Intraperitoneal administration of CQD (15 mg/kg) for 6 days was found to improve the skin lesions in the imiquimod-induced psoriatic mouse model (male C57BL/6 mice), while secretion of pro-inflammatory cytokines (IL-17 and IL-1ß) and keratinocyte proliferation were significantly suppressed by CQD. In conclusion, CQD exerted inhibitory effects on NLRP3 inflammasome activation in macrophages and decreased the severity of psoriatic response in vivo. Such findings indicate that the repurposing of the old drug, CQD, is a potential pharmacological approach for the treatment of psoriasis and other NLRP3-driven diseases.


Assuntos
Clorquinaldol , Dermatite , Psoríase , Animais , Proteínas de Transporte/metabolismo , Caspase 1/metabolismo , Clorquinaldol/efeitos adversos , Citocinas/metabolismo , Feminino , Imiquimode/toxicidade , Inflamassomos/metabolismo , Interleucina-1beta/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nucleotídeos/efeitos adversos , Nucleotídeos/metabolismo , Psoríase/induzido quimicamente , Psoríase/tratamento farmacológico , Domínio Pirina
7.
Drug Des Devel Ther ; 16: 1793-1809, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35719213

RESUMO

Purpose: Idiopathic pulmonary fibrosis is a chronic and irreversible fibrotic interstitial pneumonia of unknown etiology and therapeutic strategies are limited. Emerging evidence suggests that the continuous activation of the central nucleotide-binding oligomerization-, leucine-rich repeat-, and pyrin domain-containing protein 3 (NLRP3) inflammasome is involved in the pathogenesis of pulmonary fibrosis. Ginsenoside Rb1 (G-Rb1) is the most abundant component in the traditional Chinese herb ginseng and has anti-inflammatory and anti-fibrotic activities. The purpose of this study was to explore whether G-Rb1 exerts anti-inflammatory and anti-fibrotic activities in vivo and in vitro by suppressing the activation of the NLRP3 inflammasome and NF-κB pathway. Methods: Forty-eight male C57BL/6 mice were randomly divided into four groups (n=12/group) as follows: control, bleomycin (BLM), BLM/G-Rb1, and G-Rb1. A pulmonary fibrosis model was developed via an intratracheal injection of BLM. Six mice from each group were euthanized on days 3 and 21. The degree of pulmonary fibrosis was examined by histological evaluation and assessing α-smooth muscle actin levels. THP-1 cells were differentiated into macrophages, and stimulated by lipopolysaccharide and adenosine triphosphate. Activation of the NLRP3 inflammasome and NF-κB pathway was determined by Western blotting. Interleukin-1 beta and interleukin-18 levels were measured by ELISA. MRC-5 cells were cultured in the conditioned medium of the treated macrophages, after which markers of myofibroblasts were determined by Western blotting. Results: G-Rb1 ameliorated BLM-induced pulmonary inflammation and fibrosis in mice, and suppressed NLRP3 inflammasome activation and the NF-κB pathway in lung tissues. Moreover, interleukin-1 beta secreted after NLRP3 inflammasome activation in macrophages promoted fibroblast differentiation. G-Rb1 inhibited lipopolysaccharide- and adenosine triphosphate-induced NLRP3 inflammasome activation in macrophages and disturbed the crosstalk between macrophages and fibroblasts. Conclusion: G-Rb1 ameliorates BLM-induced pulmonary inflammation and fibrosis by suppressing NLRP3 inflammasome activation and the NF-κB pathway. Hence, G-Rb1 is a potential novel therapeutic drug for idiopathic pulmonary fibrosis.


Assuntos
Fibrose Pulmonar Idiopática , Pneumonia , Trifosfato de Adenosina/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Bleomicina/efeitos adversos , Fibrose , Ginsenosídeos , Fibrose Pulmonar Idiopática/tratamento farmacológico , Inflamassomos/metabolismo , Interleucina-1beta/metabolismo , Leucina/uso terapêutico , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nucleotídeos/metabolismo , Pneumonia/induzido quimicamente , Pneumonia/tratamento farmacológico , Pneumonia/metabolismo , Domínio Pirina , Transdução de Sinais
8.
Methods Mol Biol ; 2523: 197-207, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35759199

RESUMO

Pattern recognition receptors of innate immune cells allow the recognition of invariant microbial structures. The nucleotide-binding oligomerization domain-like receptors (NLRs) comprise 22 members, divided into 3 subfamilies. Homotypic pyrin domain (PYD) interactions were shown to mediate the interaction of inflammasome forming NLRPs with the adaptor protein ASC, bridging the interaction to caspase-1 and resulting in caspase-1-induced cytokine maturation and pyroptotic cell death. Here we describe a NLRP3PYD-mediated ASC polymerization assay that reconstitutes the transition from the NLRP3PYD nucleation seed to ASC adaptor filament elongation with recombinant proteins.


Assuntos
Proteína 3 que Contém Domínio de Pirina da Família NLR , Domínio Pirina , Caspase 1/metabolismo , Proteínas do Citoesqueleto/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Ligação Proteica
9.
J Biochem Mol Toxicol ; 36(7): e23059, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35384154

RESUMO

The kidney is highly vulnerable to cadmium-evoked oxidative injury. Galangin is a natural flavone with reported antioxidant properties. This study investigated the potential modulating activity of galangin against cadmium-induced nephrotoxicity and explored the underlining mechanisms. Western blot analysis, spectrophotometric, ELISA, and histopathological techniques were employed. The results revealed that galangin suppressed tubular injury and improved glomerular function in the cadmium-intoxicated rats as evidenced by downregulation of kidney injury molecule-1, serum creatinine, and blood urea nitrogen. Galangin reduced cadmium-evoked inflammatory response and oxidative stress as indicated by reduced levels of interleukin-1 beta and TNF-α, decreased DNA damage, and improved antioxidant potential of the renal tissues. Mechanistically, galangin suppressed the nucleotide-binding domain-like receptor pyrin domain containing 3 inflammasome and efficiently decreased caspase-1 activity in the cadmium-intoxicated rats. Equally important, it inhibited the cadmium-induced nuclear translocation of nuclear factor kappa B and upregulated nuclear factor erythroid 2-related factor 2 signaling. The results highlight the ability of galangin to attenuate cadmium-evoked nephrotoxicity and support its therapeutic implementation although clinical investigations are warranted.


Assuntos
Inflamassomos , NF-kappa B , Animais , Antioxidantes/farmacologia , Cádmio/toxicidade , Flavonoides , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Nucleotídeos , Domínio Pirina , Ratos
10.
Chem Biol Drug Des ; 100(2): 218-229, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35434894

RESUMO

Punicalagin is recorded to be a potent anti-inflammatory drug, while its effect on inflammation existing in ventilator-induced lung injury (VILI) requires further verification. Rats were pretreated with punicalagin, followed by VILI modeling. Lung histopathological examination was performed with hematoxylin-eosin staining accompanied by the lung injury score. The lung wet/dry (W/D) weight ratio and total bronchoalveolar lavage fluid (BALF) protein level were measured. After transfection with protease-activated receptor-2 (PAR2) overexpression plasmids, mouse alveolar epithelial MLE-12 cells were treated with punicalagin and then subjected to cyclic stretching. Punicalagin's cytotoxicity to MLE-12 cells were measured by MTT assay. The levels of inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6), PAR2, NLR family pyrin domain containing-3 (NLRP3), and apoptosis-associated speck-like protein containing a CARD (ASC) in the BALF, lung tissues or cells were analyzed by enzyme-linked immune-sorbent assay (ELISA), qRT-PCR or/and western blot. Punicalagin treatment attenuated VILI-induced lung histopathological changes and counteracted VILI-induced increases in the lung injury score, W/D weight ratio and total protein level in BALF. Also, punicalagin treatment counteracted in vivo VILI/cyclic stretching-induced increases in the levels of PAR2, inflammatory cytokines, NLRP3, and ASC. PAR2 overexpression potentiated the cyclic stretching-induced effects, while punicalagin treatment revoked this PAR2 overexpression-induced potentiation effect. In turn, PAR2 overexpression partly resisted the punicalagin treatment-induced counteractive effects on the cyclic stretching-induced effects. Punicalagin suppresses inflammation in VILI through PAR2 inhibition-induced inhibition of NLRP3 inflammasome activation.


Assuntos
Taninos Hidrolisáveis , Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Receptor PAR-2 , Lesão Pulmonar Induzida por Ventilação Mecânica , Animais , Citocinas/metabolismo , Taninos Hidrolisáveis/farmacologia , Inflamassomos/metabolismo , Inflamação/tratamento farmacológico , Pulmão/metabolismo , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Domínio Pirina , Ratos , Receptor PAR-2/antagonistas & inibidores , Receptor PAR-2/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/tratamento farmacológico , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/patologia
11.
Am J Pathol ; 192(6): 837-846, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35351468

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a dramatic disease without cure. The US Food and Drug Administration-approved drugs, pirfenidone and nintedanib, only slow disease progression. The clinical investigation of novel therapeutic approaches for IPF is an unmet clinical need. Nucleotide-binding oligomerization domain-like receptor or NOD-like receptors are pattern recognition receptors capable of binding a large variety of stress factors. NLR family pyrin domain-containing protein 3 (NLRP3), once activated, promotes IL-1ß, IL-18 production, and innate immune responses. Multiple reports indicate that the inflammasome NLRP3 is overactivated in IPF patients, leading to increased production of class I IL and collagens. Similarly, data from animal models of pulmonary fibrosis confirm the role of NLRP3 in the development of chronic lung injury and pulmonary fibrosis. This report provides a review of the evidence of NLRP3 activation in IPF and of NLRP3 inhibition in different animal models of fibrosis, and highlights the recent advances in direct and indirect NLRP3 inhibitors.


Assuntos
Fibrose Pulmonar Idiopática , Inflamassomos , Animais , Proteínas de Transporte/metabolismo , Humanos , Fibrose Pulmonar Idiopática/tratamento farmacológico , Inflamassomos/metabolismo , Interleucina-1beta/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Domínio Pirina
12.
Bioengineered ; 13(3): 6740-6749, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35246004

RESUMO

Sepsis brain injury (SBI) is a major cause of death in critically ill patients. The present study aimed to investigate the role of emodin in SBI development. Human astrocyte 1321N1 cells were stimulated with 100 ng/mL lipopolysaccharide (LPS) to establish an SBI model in vitro. Flow cytometry was performed to measure the cell pyroptosis. The protein expression levels of syndecan-1 (SDC-1), NLR family pyrin domain containing 3 (NLRP3), Caspase-1, and the N-terminal fragment of gasdermin D (GSDMD-N) were measured using Western blotting. Interleukin (IL)-1ß, IL-6, IL-10, and tumor necrosis factor (TNF)-α levels in cells were measured using enzyme-linked immunosorbent assay kits. The N6-methyladenosine (m6A) modification was analyzed using the methylated RNA immunoprecipitation assay. NLRP3 activator, nigericin, was used to overexpress NLRP3. LPS treatment significantly enhanced the pyroptosis in 1321N1 cells, increased the levels of TNF-α, IL-1ß, and IL-6, and decreased the levels of IL-10. The protein expression levels of NLRP3, SDC-1, GSDMD-N, and Caspase-1 were also increased. Emodin treatment decreased the levels of TNF-α, IL-1ß, IL-6, NLRP3, SDC-1, GSDMD-N, and Caspase-1, while increasing the levels of IL-10 in LPS-treated 1321N1 cells. Nigericin reversed the effects of emodin. Furthermore, emodin upregulated m6A levels in NLRP3 by increasing the expression of methyltransferase-like 3 (METTL3). Meanwhile, knockdown of METTL3 reversed the effects of emodin on the mRNA expression and stability of NLRP3. Therefore, emodin inhibits the inflammation and pyroptosis of LPS-treated 1321N1 cells by inactivating METTL3-mediated NLRP3 expression.


Assuntos
Emodina , Lipopolissacarídeos , Caspase 1/metabolismo , Caspase 1/farmacologia , Emodina/farmacologia , Humanos , Inflamação/metabolismo , Interleucina-10/farmacologia , Interleucina-6 , Lipopolissacarídeos/farmacologia , Metiltransferases/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nigericina/farmacologia , Domínio Pirina , Piroptose/fisiologia , Fator de Necrose Tumoral alfa
13.
Transpl Immunol ; 71: 101548, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35122957

RESUMO

BACKGROUND: Andrographolide (AD) has been reported to play a potential anti-arthritic role by facilitating the proliferation and inhibiting the apoptosis of chondrocytes. However, the molecular mechanism underlying the protective role of AD in osteoarthritis (OA) remains to be elucidated. METHODS: OA mice model was established via anterior cruciate ligament transection (ACLT) operation. OA cell model was established through treating mice primary chondrocytes with LPS (1 µg/mL, 24 h). Enzyme-linked immunosorbent assay (ELISA) was performed to measure the concentrations of inflammatory cytokines in the supernatant. Cell proliferation was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 5-Ethynyl-2'-deoxyuridine (EdU) assay. Cell apoptosis was evaluated by flow cytometry. The intermolecular interaction was verified by dual-luciferase reporter assay. RESULTS: AD administration reduced the infiltration of inflammatory cells in the synovial tissues of ankle joint and suppressed the inflammatory response in OA mice model in vivo. Lipopolysaccharide (LPS) stimulation suppressed the proliferation and induced the apoptosis and inflammation of chondrocytes, and AD treatment protected chondrocytes from LPS-induced dysfunction. Circular RNA (circRNA) Rap guanine nucleotide exchange factor 1 (circ_Rapgef1) overexpression attenuated AD-mediated protective effects in OA cell model. Circ_Rapgef1/microRNA-383-3p (miR-383-3p)/Nod-like receptor pyrin domain 3 (NLRP3) axis was identified in this study for the first time. Circ_Rapgef1 overexpression-mediated effects were partly reversed by the overexpression of miR-383-3p in chondrocytes. NLRP3 silencing partly overturned miR-383-3p knockdown-mediated effects in chondrocytes. Circ_Rapgef1 overexpression up-regulated the expression of NLRP3 partly by targeting miR-383-3p in chondrocytes. CONCLUSION: Circ_Rapgef1 suppressed AD-mediated protective effects in OA partly by regulating miR-383-3p/NLRP3 signaling.


Assuntos
MicroRNAs , Osteoartrite , Animais , Apoptose , Diterpenos , Lipopolissacarídeos/farmacologia , Camundongos , MicroRNAs/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteínas NLR , Osteoartrite/tratamento farmacológico , Domínio Pirina
14.
J Biol Chem ; 298(3): 101566, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35007535

RESUMO

ASC is an essential adaptor of the inflammasome, a micrometer-size multiprotein complex that processes proinflammatory cytokines. Inflammasome formation depends on ASC self-association into large assemblies via homotypic interactions of its two death domains, PYD and CARD. ASCb, an alternative splicing isoform, activates the inflammasome to a lesser extent compared with ASC. Thus, it has been postulated that adaptor isoforms differentially regulate inflammasome function. At the amino acid level, ASC and ASCb differ only in the length of the linker connecting the two death domains. To understand inflammasome regulation at the molecular level, we investigated the self-association properties of ASC and ASCb using real-time NMR, dynamic light scattering (DLS), size-exclusion chromatography, and transmission electron microscopy (TEM). The NMR data indicate that ASC self-association is faster than that of ASCb; a kinetic model for this oligomerization results in differing values for both the reaction order and the rate constants. Furthermore, DLS analysis indicates that ASC self-associates into more compact macrostructures compared with ASCb. Finally, TEM data show that ASCb has a reduced tendency to form densely packed filaments relative to ASC. Overall, these differences can only be explained by an effect of the linker length, as the NMR results show structural equivalence of the PYD and CARD in both proteins. The effect of linker length was corroborated by molecular docking with the procaspase-1 CARD domain. Altogether, our results indicate that ASC's faster and less polydisperse polymerization is more efficient, plausibly explaining inflammasome activation differences by ASC isoforms at the molecular level.


Assuntos
Proteínas Adaptadoras de Sinalização CARD , Inflamassomos , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Caspase 1/metabolismo , Domínio de Ativação e Recrutamento de Caspases , Inflamassomos/metabolismo , Simulação de Acoplamento Molecular , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Isoformas de Proteínas , Domínio Pirina
15.
Eur J Ophthalmol ; 32(5): 3058-3063, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35068231

RESUMO

PURPOSE: This study aimed to explore the pterygium formation and recurrence, by detecting the expression of Nod-like receptor pyrin domain 3 (NLRP3) and Nod-like receptor pyrin domain 6 (NLRP6) in pterygium and evaluate the correlation between NLRP3 and NLRP6 in pterygium. METHODS: In this prospective study, the expression levels of NLRP3 and NLRP6, with their related effectors, were evaluated in primary pterygium (n = 40) and recurrent pterygium (n = 32) tissue samples and compared with normal conjunctiva (n = 11) tissue samples by immunohistochemistry. RESULTS: Compared to the normal conjunctiva group, the expression levels of NLRP3, caspase-1, IL-18, and IL-1ß, were significantly higher, and NLRP6 showed an expression that was significantly lower in pterygium tissue samples (P < 0.05, respectively). Compared to the primary pterygium group, the expression levels of NLRP3, caspase-1, IL-18 and IL-1ß were significantly higher, and NLRP6 showed an expression that was significantly lower in recurrent pterygium tissue samples (P < 0.05, respectively).There was a negative correlation between NLRP3 expression and NLRP6 expression in normal conjunctival (r = -0.739, P = 0.009) and pterygium (r = -0.533, P = 0.000). CONCLUSIONS: NLRP3 and NLRP6 may be involved in the formation and recurrence of pterygium. NLRP6 may play an anti-inflammatory role in normal conjunctival tissue to maintain conjunctival homeostasis.


Assuntos
Pterígio , Caspases/metabolismo , Túnica Conjuntiva/anormalidades , Túnica Conjuntiva/metabolismo , Humanos , Interleucina-18/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteínas NLR/metabolismo , Estudos Prospectivos , Pterígio/metabolismo , Domínio Pirina , Recidiva
16.
Phytomedicine ; 95: 153865, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34856474

RESUMO

BACKGROUND: Tibial dyschondroplasia (TD) is a common disease characterized by proliferation and the deterioration of growth plate's chondrocytes due to widespread utilization of thiram in the agriculture and industrial sector. PURPOSE: In recent years, Nod-like receptor pyrin domain 3 (NLRP3) inflammasome has become a dilemma in the occurrence of many diseases. According to many research investigations, NLRP3 inflammasome has been linked to various diseases caused by pesticides and environmental toxins. Its involvement in such conditions opens up new treatment approaches. However, the role of the NLRP3 inflammasome in the development of TD is not fully understood under the impact of chlorogenic acid (CGA). METHODS: Chondrocytes were cultured with our previously developed methodology from growth plates. After morphological and molecular identification, chondrocytes were split into different groups to investigate the efficacy of chlorogenic acid. Cell apoptosis was determined through flow cytometry and Tunnel assay. Furthermore, RT-qPCR, immunofluorescence, and western blotting techniques were used to check marker genes and proteins expression. RESULTS: In thiram-induced TD, Bax/Bak activation persuade a parallel pathway, mediated by the NLRP3 base inflammasome. It is worth mentioning that the apoptotic executioners (caspase-3 and caspase-7) act upstream for inflammasome. Furthermore, chondrocytes' ability to undergo mitochondrial apoptosis was governed by anti-apoptotic members, e.g., Bcl-2 and Bcl-xl. Equilibrium of these anti-apoptotic proteins ensured appropriate regulation of apoptosis during the development and survival of chondrocytes. CONCLUSION: Chondrocytes have ability to undergo Bax/Bak-mediated apoptosis and generate pro-inflammatory signals, e.g., NLRP3 in thiram-induced TD. So, the Nod-like receptor pyrin domain 3 is the potential target to eliminate TD at all stages of pathology, while drugs, e.g., CGA, can significantly improve chondrocytes' survival by targeting these pro-inflammatory signals.


Assuntos
Ácido Clorogênico/farmacologia , Condrócitos/efeitos dos fármacos , Inflamassomos , Tiram , Animais , Galinhas , Proteína 3 que Contém Domínio de Pirina da Família NLR , Domínio Pirina , Proteína Killer-Antagonista Homóloga a bcl-2 , Proteína X Associada a bcl-2
17.
Anal Biochem ; 638: 114510, 2022 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-34863712

RESUMO

The NLRP3 inflammasome is a key macromolecular complex of the innate immune system that activates the inflammatory signalling cascade in response to a wide range of stimuli. Structural studies have shown that the intracellular cytosolic receptor NLRP3 oligomerizes upon stimulation and serves as a scaffold to form the ASC filaments necessary for procaspase-1 activation. Despite the abundant structural evidences on NLRP3 inflammasome, the interactions of the NLRP3 Pyrin domain and its functional relevance are poorly understood. In this study, the split luciferase complementation assay is used as an alternative approach to investigate NLRP3PYD-NLRP3PYD interactions during inflammasome formation. Since the homotypic NLRP3 interaction is mainly based on electrostatic interactions, a phosphomimetic residue (S5) at the interface of the NLRP3PYDs interactions has been mutated to show a disruptive effect on luciferase activity. According to the results presented, the designed biosensor was able to monitor the NLRP3PYD-NLRP3PYD interaction in vitro. The current reporter assay not only provides a specific NLRP3PYD-NLRP3PYD assay to study the PYD-PYD interaction in vitro, but also provides a suitable system for screening chemicals and drugs to identify activators and inhibitors of NLRP3.


Assuntos
Técnicas Biossensoriais , Inflamassomos/metabolismo , Inflamação/metabolismo , Luciferases/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Humanos , Inflamassomos/química , Proteína 3 que Contém Domínio de Pirina da Família NLR/química , Domínio Pirina
18.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(5): 788-795, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34728041

RESUMO

Alzheimer's disease(AD)is a chronic neurodegenerative disease whose cause remains unclear.The ß-amyloid plaques in the brain are one of the major pathological features of AD.However,the drugs targeting extracellular ß-amyloid plaques have failed to cure the disease.Innate immunity and neuroinflammation play a role in the pathogenesis and progression of AD.As the macrophages existing in the central nervous system,microglia are related with extracellular ß-amyloid deposition,intracellular neurofibrillary tangle formation,and neuron injury.Accumulating evidence demonstrates that the activation of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome in microglia plays a role in AD,suggesting new therapeutic target for AD in this signaling pathway.This article reviewed the studies about the activation and regulation of NLRP3 inflammasome in the pathogenesis and progression of AD as well as the development of AD therapies targeting this pathway,aiming to provide reference for further studies in this field.


Assuntos
Doença de Alzheimer , Doenças Neurodegenerativas , Humanos , Inflamassomos , Microglia , Proteína 3 que Contém Domínio de Pirina da Família NLR , Nucleotídeos , Domínio Pirina
19.
Hum Exp Toxicol ; 40(12_suppl): S540-S552, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34715758

RESUMO

Ischemic stroke is a leading cause of death and long-term disability worldwide. The aim of this study is to explore the potential function of ephedrine in ischemic stroke and the underlying molecular mechanism. A middle cerebral artery occlusion (MCAO) rat model was established. The potential effects of ephedrine on MCAO rats and LPS-stimulated BV2 microglial cells were evaluated. Ephedrine reduced the infarct volume, cell apoptosis, brain water content, neurological score, and proinflammatory cytokines (TNF-α and IL-1ß) production in MCAO rats. Ephedrine treatment also suppressed TNF-α and IL-1ß production and NOD-like receptor pyrin domain 3 (NLRP3) inflammasome activation in BV2 microglial cells. The expression of NLRP3, caspase-1, and IL-1ß was suppressed by ephedrine. Moreover, ephedrine treatment increased the phosphorylation of Akt and GSK3ß and nuclear NRF2 levels in LPS-treated BV2 microglial cells. Meanwhile, LY294002 attenuated the inhibitory effects of ephedrine on NLRP3 inflammasome activation and TNF-α and IL-1ß production. In addition, the level of pAkt was increased, while NLRP3, caspase-1, and IL-1ß were decreased by ephedrine treatment in MCAO rats. In conclusion, ephedrine ameliorated cerebral ischemia injury via inhibiting NLRP3 inflammasome activation through the Akt/GSK3ß/NRF2 pathway. Our results revealed a potential role of ephedrine in ischemic stroke treatment.


Assuntos
Isquemia Encefálica/prevenção & controle , Efedrina/farmacologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Inflamassomos/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Domínio Pirina , Animais , Isquemia Encefálica/metabolismo , Masculino , Fosforilação , Ratos , Ratos Sprague-Dawley
20.
J Dairy Sci ; 104(11): 11973-11982, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34454753

RESUMO

Free fatty acids (FFA)-induced hepatic inflammation agravates liver injury and metabolic dysfunction in dairy cows with ketosis or fatty liver. Under stressful conditions, autophagy is generally considered as a cell protection mechanism, but whether the FFA-induced inflammatory and stress effect on hepatocytes involves an autophagy response is not well known. Thus, the objective of this study was to investigate the effects of FFA on autophagy and the role of autophagy in the activation of NF-κB (nuclear factor kappa B) signaling and NLRP3 (NLR family pyrin domain containing 3) inflammasome in calf hepatocytes. Calf hepatocytes were isolated from 3 healthy Holstein female new-born calves (1 d of age, 30-40 kg) and exposed to various concentrations of FFA (0, 0.3, 0.6, or 1.2 mM) after treatment with or without the autophagy inhibitor chloroquine (CQ) or the autophagy activator rapamycin. Expression of autophagy markers, LC3 (microtubule-associated protein 1 light chain 3) and p62 (sequestosome 1), NF-κB signaling, and NLRP3 inflammasome-related molecules were analyzed via western blot and quantitative real-time PCR. Results revealed that 0.6 and 1.2 mM FFA activated NF-κB signaling and NLRP3 inflammasome as indicated by an elevated ratio of p-NF-κB/NF-κB, protein abundance of NLRP3 and CASP1 (caspase 1), activity of CASP1, and mRNA abundance of IL1B and IL18. In addition, hepatocyte treated with 0.6 and 1.2 mM FFA or autophagy inhibitor CQ displayed increased protein abundance of p62 and LC3-II. Moreover, there was no difference in protein abundance of p62 and LC3-II between calf hepatocytes treated with 1.2 mM FFA and 1.2 mM FFA plus CQ, indicating that FFA inhibits autophagic activity in calf hepatocytes. Treatment with CQ led to overactivation of NF-κB signaling and NLRP3 inflammasome. Furthermore, CQ plus 1.2 mM FFA aggravated FFA-induced inflammation. In contrast, induction of autophagy by rapamycin ameliorated the FFA-activated NF-κB signaling and NLRP3 inflammasome as demonstrated by a lower ratio of p-NF-κB/NF-κB, protein abundance of NLRP3 and CASP1, activity of CASP1, and mRNA abundance of IL1B and IL18. Overall, inhibition of autophagy exacerbated, whereas induction of autophagy alleviated, FFA-induced inflammatory processes in calf hepatocytes, suggesting that impairment of autophagy might be partly responsible for hepatic inflammation and subsequent liver injury in dairy cows with ketosis or fatty liver. As such, regulation of autophagy may be an effective therapeutic strategy for controlling overt inflammatory responses in vivo.


Assuntos
Inflamassomos , NF-kappa B , Animais , Autofagia , Bovinos , Ácidos Graxos não Esterificados , Feminino , Hepatócitos/metabolismo , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR , Gravidez , Domínio Pirina
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...