AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid (PUFA)-enriched diet. Pr...AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid (PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids (SFAs) or PUFAs as well as combined with lipopolysaccharide (LPS). The expression of NOD-like receptor protein 3 (NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B (NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: High-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid (PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid (DHA) had the potential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a high-fat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but DHA decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION: Hepatic NLRP3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB.展开更多
OBJECTIVE:To investigate the impact of Yemazhui(Herba Eupatorii Lindleyani,HEL)against lipopolysaccharide(LPS)-induced acute lung injury(ALI)and explore its underlying mechanism in vivo.METHODS:The chemical constituen...OBJECTIVE:To investigate the impact of Yemazhui(Herba Eupatorii Lindleyani,HEL)against lipopolysaccharide(LPS)-induced acute lung injury(ALI)and explore its underlying mechanism in vivo.METHODS:The chemical constituents of HEL were analyzed by ultra-high performance liquid chromatographyquadrupole time-of-flight mass spectrometry method.Then,HEL was found to suppress LPS-induced ALI in vivo.Six-week-old male Sprague-Dawley rats were randomly divided into 6 groups:control,LPS,Dexamethasone(Dex),HEL low dose 6 g/kg(HEL-L),HEL medium dose 18 g/kg(HEL-M)and HEL high dose 54 g/kg(HEL-H)groups.The model rats were intratracheally injected with 3 mg/kg LPS to establish an ALI model.Leukocyte counts,lung wet/dry weight ratio,as well as myeloperoxidase(MPO)activity were determined followed by the detection with hematoxylin and eosin staining,enzyme linked immunosorbent assay,quantitative real time polymerase chain reaction,western blotting,immunohistochemistry,and immunofluorescence.Besides,to explore the effect of HEL on ALI-mediated intestinal flora,we performed 16s rRNA sequencing analysis of intestinal contents.RESULTS:HEL attenuated LPS-induced inflammation in lung tissue and intestinal flora disturbance.Mechanism study indicated that HEL suppressed the lung coefficient and wet/dry weight ratio of LPS-induced ALI in rats,inhibited leukocytes exudation and MPO activity,and improved the pathological injury of lung tissue.In addition,HEL reduced the expression of tumor necrosis factoralpha,interleukin-1beta(IL-1β)and interleukin-6(IL-6)in bronchoalveolar lavage fluid and serum,and inhibited nuclear displacement of nuclear factor kappa-B p65(NF-κBp65).And 18 g/kg HEL also reduced the expression levels of toll-like receptor 4(TLR4),myeloid differentiation factor 88,NF-κBp65,phosphorylated inhibitor kappa B alpha(phospho-IκBα),nod-like receptor family pyrin domain-containing 3 protein(NLRP3),IL-1β,and interleukin-18(IL-18)in lung tissue,and regulated intestinal flora disturbance.CONCLUSIONS:In summary,our findings revealed that HEL has a protective effect on LPS-induced ALI in rats,and its mechanism may be related to inhibiting TLR4/NF-κB/NLRP3 signaling pathway and improving intestinal flora disturbance.展开更多
Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,mo...Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,most studies have focused mainly on whether diabetes increases the NLRP3 inflammasome and associated pyroptosis in the heart of type 1 or type 2 diabetic rodent models,and whether various medications and natural products prevent the development of DCM,associated with decreased levels of cardiac NLRP3 inflammasome and pyroptosis.The direct link of NLRP3 inflammasome and associated pyroptosis to the pathogenesis of DCM remains unclear based on the limited evidence derived from the available studies,with the approaches of NLRP3 gene silencing or pharmaceutical application of NLRP3 specific inhibitors.We thus emphasize the requirement for more systematic studies that are designed to provide direct evidence to support the link,given that several studies have provided both direct and indirect evidence under specific conditions.This editorial emphasizes that the current investigation should be circumspect in its conclusion,i.e.,not overemphasizing its role in the pathogenesis of DCM with the fact of only significantly increased expression or activation of NLRP3 inflammasome and pyroptosis in the heart of diabetic rodent models.Only clear-cut evidence-based causative roles of NLRP3 inflammasome and pyroptosis in the pathogenesis of DCM can help to develop effective and safe medications for the clinical management of DCM,targeting these biomarkers.展开更多
Introduction Inflammatory bowel diseases(IBD),such as Crohn’s disease(CD)and ulcerative colitis(UC),are a group of chronic inflammatory disorders of the gastrointestinal tract[1-2].The symptoms of IBD include abdomin...Introduction Inflammatory bowel diseases(IBD),such as Crohn’s disease(CD)and ulcerative colitis(UC),are a group of chronic inflammatory disorders of the gastrointestinal tract[1-2].The symptoms of IBD include abdominal pain,diarrhea,and bloody stool.IBD affects a patient’s quality of life severely,due in part to its frequent recurrence.Colorectal cancer(CRC)is a malignancy in the colon or rectum with symptoms including bloody stool,changes in展开更多
Background:Dermatomyositis(DM)and polymyositis(PM)are common inflammatory myopathies whose immunopathogenic mechanisms remain poorly understood.The NOD-like receptor family,pyrin domain containing 3(NLRP3)inflammasome...Background:Dermatomyositis(DM)and polymyositis(PM)are common inflammatory myopathies whose immunopathogenic mechanisms remain poorly understood.The NOD-like receptor family,pyrin domain containing 3(NLRP3)inflammasome is a type of cytoplasmic multiprotein inflammasome and is responsible for the activation of inflammatory reactivations.Responding to a wide range of exogenous and endogenous microbial or sterile stimuli,NLRP3 inflammasomes can cleave pro-caspase-1 into active caspase-1,which processes the pro-infammatory cytokines pro-interleukin(IL)-1βand pro-IL-18 into active and secreted IL-1βand I L-18.The NLRP3 inflammasome is implicated in infectious and sterile inflammatory diseases.However,it remains unclear whether it is involved in the pathogenesis of DM/PM,which we aim to address in our research.Methods:In this study,22 DM/PM patients and 24 controls were recruited.The protein and RNA expression of IL-113,IL-18,NLRP3,and caspase-1 in serum and muscle samples were tested and compared between the two groups.Results:The serum IL-1βand IL-18 levels were significantly higher in DM/PM patients than those in the controls by enzyme linked immunosorbent assay(EL1SA,DM vs.control,25.02±8.29 ng/ml vs.16.49±3.30 ng/ml,P〈0.001;PM vs.control,26.49±7.79 ng/ml vs.16.49±3.30 ng/ml,P〈0.001).Moreover,the real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR)showed that DM/PM patients exhibited higher RNA expression of IL-lβ,IL-18,and NLRP3 in the muscle(for IL-1β,DM vs.control,P 0.0012,PM vs.control,P=0.0021;for IL-18,DM vs.control,P=0.0045,PM vs.control,P 0.0031;for NLRP3,DM vs.control,P=0.0017,PM vs.control,P 0.0006).Moreover,the protein expression of NLRP3 and caspase-1 in muscle samples of DM/PM patients were also significantly elevated compared to that in the muscles of the controls.Conclusions:Our findings demonstrate that the NLRP3 inflammasome is implicated in the pathogenesis of DM/PM.High NLRP3 expression led to elevated levels of IL-l13 and IL-18 and could be one of the factors promoting disease progress.展开更多
Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms invo...Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms involving skin, joints, central nervous system, and eyes. It encompasses a spectrum of three clinically overlapping autoinflammatory syndromes including familial cold autoinflammatory syndrome, Muckle-Wells syndrome （MWS）, and neonatal-onset multisystem inflammatory disease. CAPS is associated with gain-of-function missense mutations in NOD-like receptor family pyrin domain-containing protein 3 （NLRP3）, the gene encoding NLRP3. Moreover, most mutations leading to MWS occurred in exon 3 ofNLRP3 gene. Here, we reported a novel mutation occurred in exon 1 ofNLRP3 gene in an MWS patient and attempted to explore the pathogenic mechanism. Methods： Genetic sequence analysis of NLRP3 was performed in an MWS patient who presented with periodic lever, arthralgia, and multiform skin lesions. NLRP3 was also analyzed in this patient＇s parents and 50 healthy individuals. Clinical examinations including X-ray examination, skin biopsy, bone marrow aspiration smear, and blood test of C-reactive protein （CRP）, erythrocyte sedimentation rate （ESR）, serum levels oflL-1β, immunoglobulin E （lgE）, antineutrophil cytoplasmic antibodies, antinuclear antibodies, and extractable nuclear antigen were also analyzed. The protein structure of mutant NLRP3 inflammasome was calculated by SWISS-MODEL software. Proteins of wild type and mutant components ofNLRP3 inflammasome were expressed and purified, and the interaction abilities between these proteins were tested by surface plasmon resonance （SPR） assay. Results： X-ray examination showed no abnormality in the patient＇s knees. Laboratory tests indicated an elevation of CRP （233.24 nag/L） and ESR （67 mm/h） when the patient had fever. Serum IL-1β increased to 24.37 pg/ml, and serum lgE was higher than 2500.00 IU/ml. Other blood tests were normal. Bone marrow aspiration smear was normal. A novel point mutation c.92A〉T in exon 1 of NLRP3 gene was identified, which caused a p.D31V mutation in pyrin domain （PYD） of NLRP3. SPR assay showed that this point mutation may strengthen the interaction between the PYD of NLRP3 and the PYD of the apoptosis-associated speck-like protein. The mutation c.92A〉T in exon 1 of the NLRP3 gene was not lbund in the patient＇s parents and 50 healthy individuals. Conclusions： The rnutation c.92A〉T in exon 1 of the NLRP3 gene is a novel mutation associated with MWS. The p.D31V mutation might promote the activation ofNLRP3 inflammasome and induce MWS in this patient.展开更多
Objective:To investigate the effect of curcumol on NOD-like receptor thermoprotein domain 3(NLRP3)inflammasomes,and analyze the mechanism underlying curcumol against liver fibrosis.Methods:Thirty Kunming mice were div...Objective:To investigate the effect of curcumol on NOD-like receptor thermoprotein domain 3(NLRP3)inflammasomes,and analyze the mechanism underlying curcumol against liver fibrosis.Methods:Thirty Kunming mice were divided into a control group,a model group and a curcumol group according to a random number table,10 mice in each group.Mice were intraperitoneally injected with 40% carbon tetrachloride(CCl4:peanut oil,2:3 preparation)at 5 m L/kg for 6 weeks,twice a week,for developing a liver fibrosis model.The mice in the control group were given the same amount of peanut oil,twice a week for 6 weeks.The mice in the curcumol group were given curcumol(30 m L/kg)intragastrically,and the mice in the model and control groups were given the same amount of normal saline,once a day for 6 weeks.Changes in liver structure were observed by hematoxylin and eosin(HE)and Masson staining.Liver function,liver fiber indices,and the expression of interleukin(IL)-10 and tumor necrosis factor-α(TNF-α)levels were determined by automatic biochemical analyzer and enzyme linked immunosorbent assay kit.Immunoblotting and reverse transcription-quantitative PCR(RT-qPCR)were performed to detect the expression of NLRP3 inflammasome-related molecules,TGF-β and collagen.Results:HE and Masson staining results showed that the hepatocytes of the model group were arranged irregularly with pseudo-lobular structure and a large amount of collagen deposition.The mice in the curcumol group had a significant decrease in liver function and liver fibers indices compared with the model group(P<0.05);RT-qPCR and Western blot results reveal that,in the curcumol group,the mRNA and protein expression levels of NLRP3,IL-1β,Caspase 1 and gasdermin D decreased significantly compared with the model group(P<0.05);immunohistochemical results showed that in the curcumol group,the protein expression levels of NLRP3 and IL-1β decreased significantly compared with the model group(P<0.05).Conclusion:A potential anti-liver fibrosis mechanism of curcumol may be associated with the inhibition of NLRP3 inflammasomes and decreasing the downstream inflammatory response.展开更多
Background: Idiosyncratic drug-induced liver injury(IDILI) is a serious side effect of drugs, Epimedii Folium(EF) is unequivocally implicated in idiosyncratic liver injury onset, potentially due to its ability to pert...Background: Idiosyncratic drug-induced liver injury(IDILI) is a serious side effect of drugs, Epimedii Folium(EF) is unequivocally implicated in idiosyncratic liver injury onset, potentially due to its ability to perturb the NOD-like receptor family pyrin domain containing 3(NLRP3) inflammasome. Fructus Ligustri Lucidi(FLL), a frequently used medicinal combination with EF, has not yet been investigated for its ability to ameliorate EF-associated hepatotoxicity. Aims and Objectives: Study on the mechanism of compatibility of FLL to alleviate liver injury caused by EF. Materials and Methods: Western blot was used to determine the expression of related proteins, ELISA was used to detect the secretion of related inflammatory factors IL-1β, IL-18, IL-6 and TNF-α, liver injury indexes were detected and liver pathological tissue staining was used to evaluate the liver injury. Results: Our results demonstrated that EF exerted a particular augmenting effect on the stimulation of the NLRP3 inflammasome mediated by nigericin or ATP, whereas FLL suppressed the NLRP3 inflammasome stimulation. Furthermore, an equal EF to FLL ratio significantly reduced the stimulatory effects of EF. Moreover, EF has the potential to induce hepatic injury and augment pro-inflammatory cytokine synthesis in rats subjected to LPS. However, when combined with FLL, the detrimental effects of EF were mitigated. Conclusions: FLL possesses the capacity to attenuate EF-associated hepatotoxicity by suppressing EF-triggered NLRP3 inflammasome activation. Thus, FLL holds promise for improving the clinical safety profile of EF, shedding light on the potential of compatibility and detoxification theories in traditional Chinese medicine.展开更多
目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细...目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细胞分为8组:(i)正常对照组:正常培养的神经元细胞;(ii)氧-糖剥夺/复氧(OGD/R)组:采用氧-糖剥夺/复氧法模拟脑缺血再灌注损伤;(iii)OGD/R+SGB组:OGD/R联合麻醉药0.5%布比卡因用于体外模拟SGB;(iv)OGD/R+SGB+TUG1过表达阴性对照组:OGD/R联合布比卡因并联合TUG1过表达阴性对照质粒转染细胞;(v)OGD/R+SGB+TUG1过表达组:OGD/R联合布比卡因并联合TUG1过表达质粒转染细胞;(vi)OGD/R+SGB+TUG1过表达+MCC950组:OGD/R联合布比卡因、TUG1过表达质粒转染及NLRP3抑制剂MCC950处理细胞;(vii)OGD/R+TUG1过表达组:OGD/R联合TUG1过表达质粒转染细胞;(viii)OGD/R+MCC950组:OGD/R联合NLRP3抑制剂MCC950处理细胞。进一步通过实时定量PCR(Quantitative Real Time PCR,qRTPCR)实验检测细胞中lncRNATUG1的表达;利用Western blot法检测细胞中NLRP3、微管相关蛋白1轻链3(LC3)-I、LC3-II、自噬相关基因5(Atg5)、苄氯素1(beclin1)、自噬接头蛋白(p62)、溶酶体相关膜蛋白1(LAMP1)的表达水平;利用透射电镜(TEM)检测自噬溶酶体的数量;并用酶联免疫吸附测定(ELISA)法检测细胞培养上清中白细胞介素(IL)-1β、IL-6、IL-18、肿瘤坏死因子(TNF)-α的含量。结果与正常对照组相比,OGD/R组lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05)。与OGD/R组相比,OGD/R+SGB组的上述指标均显著下调(P<0.05)。与OGD/R+SGB+TUG1过表达阴性对照组相比,OGD/R+SGB+TUG1过表达组的lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05),然而,加入NLRP3的抑制剂MCC950后,除lncRNA TUG1外其余指标均显著下调(^(均)P<0.05)。另外,与OGD/R组比,OGD/R+TUG1过表达组的上述指标进一步上调(^(均)P<0.05)。与OGD/R组比,OGD/R+MCC950组则抑制了除lncRNA TUG1外的其余指标(^(均)P<0.05)。结论星状神经节阻滞通过调节lncRNA TUG1-NLRP3轴有效减轻体外脑缺血再灌注损伤引起的炎症反应和自噬溶酶体形成,提示其可能作为治疗缺血性脑损伤的潜在策略。展开更多
Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic...Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic effects of polydatin(20 and 40 mg/kg)were evaluated in this asthmatic mouse model.To assess the underlying mechanisms,Bronchial Epithelium Adenovirus 12-SV402B(BEAS-2B)cells were cocultured with Tohoku Hospital for Pediatrics-1(THP-1)macrophages,in which toll-like receptor 4(TLR4)was either overexpressed or knocked down,and subsequently stimulated with lipopoly-saccharide(LPS)and ATP.THP-1 cells underwent a 1-h pretreatment with polydatin(50 and 100μmol/L),Class Lipid Inhibitor-095(CLI-095,TLR4 inhibitor,1μg/mL),or A438079(P2X7R antagonist,10μmol/L)prior to LPS/ATP challenge.Results:Findings from Western blotting,enzyme-linked immunosorbent assay,flow cytometry,real-time polymerase chain reaction,and immunofluorescence assays demonstrated that modulating TLR4 expression significantly altered interleukin-1β(IL-1β)secretion from THP-1 macrophages and mitochondrial reactive oxygen species(mtROS)production in BEAS-2B ECs.In the mouse asthma model,polydatin significantly alleviated airway inflammation,oxidative stress,and apoptosis,likely by interfering with TLR4/P2X7R-mediated signaling and suppressing the activation of the NOD-like receptor protein inflammasome.Additionally,polydatin significantly reduced IL-1βand IL-18 levels and inhibited the infiltration of macrophages and eosinophils.Correspondingly,polydatin significantly attenuated TLR4/P2X7R signaling in THP-1 cells stimulated with ATP and LPS,thereby reducing IL-1βand IL-18 secretion,calcium influx,mtROS production,and apoptosis in BEAS-2B ECs.Conclusions:Polydatin is a promising therapeutic candidate for asthma,possibly by targeting macrophage-epithelium cross-talk via the TLR4/P2X7R axis.Future formulations as capsules or sprays may effectively alleviate airway inflammation and remodeling.展开更多
基金Supported by The National Natural Science Foundation of ChinaNO.81170374 and NO.81470842 to Hua J
文摘AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid (PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids (SFAs) or PUFAs as well as combined with lipopolysaccharide (LPS). The expression of NOD-like receptor protein 3 (NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B (NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: High-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid (PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid (DHA) had the potential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a high-fat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but DHA decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION: Hepatic NLRP3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB.
基金Natural Science Foundation Project of Chongqing Municipality:a Metabolome-based Study on the Protective Mechanism of Yemazhui(Herba Eupatorii Lindleyani)Sesquiterpene Lactones Against Acute Lung Injury(No.cstc2021jcyj-msxmX0365)Science and Technology Research Program of Chongqing Municipal Education Commission:a Cytokine Storm-based Study of the Protective Effect of Yemazhui(Herba Eupatorii Lindleyani)Extract Intervention on COVID-19 Lung Injury(No.KJZD-K202215101)。
文摘OBJECTIVE:To investigate the impact of Yemazhui(Herba Eupatorii Lindleyani,HEL)against lipopolysaccharide(LPS)-induced acute lung injury(ALI)and explore its underlying mechanism in vivo.METHODS:The chemical constituents of HEL were analyzed by ultra-high performance liquid chromatographyquadrupole time-of-flight mass spectrometry method.Then,HEL was found to suppress LPS-induced ALI in vivo.Six-week-old male Sprague-Dawley rats were randomly divided into 6 groups:control,LPS,Dexamethasone(Dex),HEL low dose 6 g/kg(HEL-L),HEL medium dose 18 g/kg(HEL-M)and HEL high dose 54 g/kg(HEL-H)groups.The model rats were intratracheally injected with 3 mg/kg LPS to establish an ALI model.Leukocyte counts,lung wet/dry weight ratio,as well as myeloperoxidase(MPO)activity were determined followed by the detection with hematoxylin and eosin staining,enzyme linked immunosorbent assay,quantitative real time polymerase chain reaction,western blotting,immunohistochemistry,and immunofluorescence.Besides,to explore the effect of HEL on ALI-mediated intestinal flora,we performed 16s rRNA sequencing analysis of intestinal contents.RESULTS:HEL attenuated LPS-induced inflammation in lung tissue and intestinal flora disturbance.Mechanism study indicated that HEL suppressed the lung coefficient and wet/dry weight ratio of LPS-induced ALI in rats,inhibited leukocytes exudation and MPO activity,and improved the pathological injury of lung tissue.In addition,HEL reduced the expression of tumor necrosis factoralpha,interleukin-1beta(IL-1β)and interleukin-6(IL-6)in bronchoalveolar lavage fluid and serum,and inhibited nuclear displacement of nuclear factor kappa-B p65(NF-κBp65).And 18 g/kg HEL also reduced the expression levels of toll-like receptor 4(TLR4),myeloid differentiation factor 88,NF-κBp65,phosphorylated inhibitor kappa B alpha(phospho-IκBα),nod-like receptor family pyrin domain-containing 3 protein(NLRP3),IL-1β,and interleukin-18(IL-18)in lung tissue,and regulated intestinal flora disturbance.CONCLUSIONS:In summary,our findings revealed that HEL has a protective effect on LPS-induced ALI in rats,and its mechanism may be related to inhibiting TLR4/NF-κB/NLRP3 signaling pathway and improving intestinal flora disturbance.
文摘Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,most studies have focused mainly on whether diabetes increases the NLRP3 inflammasome and associated pyroptosis in the heart of type 1 or type 2 diabetic rodent models,and whether various medications and natural products prevent the development of DCM,associated with decreased levels of cardiac NLRP3 inflammasome and pyroptosis.The direct link of NLRP3 inflammasome and associated pyroptosis to the pathogenesis of DCM remains unclear based on the limited evidence derived from the available studies,with the approaches of NLRP3 gene silencing or pharmaceutical application of NLRP3 specific inhibitors.We thus emphasize the requirement for more systematic studies that are designed to provide direct evidence to support the link,given that several studies have provided both direct and indirect evidence under specific conditions.This editorial emphasizes that the current investigation should be circumspect in its conclusion,i.e.,not overemphasizing its role in the pathogenesis of DCM with the fact of only significantly increased expression or activation of NLRP3 inflammasome and pyroptosis in the heart of diabetic rodent models.Only clear-cut evidence-based causative roles of NLRP3 inflammasome and pyroptosis in the pathogenesis of DCM can help to develop effective and safe medications for the clinical management of DCM,targeting these biomarkers.
基金supported by Funds from Talents’Start-up Fund of Gannan Medical University(QD201404,LIU Zhiping)Natural Science Foundation of Jiangxi Province(20151512040424,LIU Zhi-ping)
文摘Introduction Inflammatory bowel diseases(IBD),such as Crohn’s disease(CD)and ulcerative colitis(UC),are a group of chronic inflammatory disorders of the gastrointestinal tract[1-2].The symptoms of IBD include abdominal pain,diarrhea,and bloody stool.IBD affects a patient’s quality of life severely,due in part to its frequent recurrence.Colorectal cancer(CRC)is a malignancy in the colon or rectum with symptoms including bloody stool,changes in
基金This work was supported by a grant from the National Natural Science Foundation of China(No.81271399).
文摘Background:Dermatomyositis(DM)and polymyositis(PM)are common inflammatory myopathies whose immunopathogenic mechanisms remain poorly understood.The NOD-like receptor family,pyrin domain containing 3(NLRP3)inflammasome is a type of cytoplasmic multiprotein inflammasome and is responsible for the activation of inflammatory reactivations.Responding to a wide range of exogenous and endogenous microbial or sterile stimuli,NLRP3 inflammasomes can cleave pro-caspase-1 into active caspase-1,which processes the pro-infammatory cytokines pro-interleukin(IL)-1βand pro-IL-18 into active and secreted IL-1βand I L-18.The NLRP3 inflammasome is implicated in infectious and sterile inflammatory diseases.However,it remains unclear whether it is involved in the pathogenesis of DM/PM,which we aim to address in our research.Methods:In this study,22 DM/PM patients and 24 controls were recruited.The protein and RNA expression of IL-113,IL-18,NLRP3,and caspase-1 in serum and muscle samples were tested and compared between the two groups.Results:The serum IL-1βand IL-18 levels were significantly higher in DM/PM patients than those in the controls by enzyme linked immunosorbent assay(EL1SA,DM vs.control,25.02±8.29 ng/ml vs.16.49±3.30 ng/ml,P〈0.001;PM vs.control,26.49±7.79 ng/ml vs.16.49±3.30 ng/ml,P〈0.001).Moreover,the real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR)showed that DM/PM patients exhibited higher RNA expression of IL-lβ,IL-18,and NLRP3 in the muscle(for IL-1β,DM vs.control,P 0.0012,PM vs.control,P=0.0021;for IL-18,DM vs.control,P=0.0045,PM vs.control,P 0.0031;for NLRP3,DM vs.control,P=0.0017,PM vs.control,P 0.0006).Moreover,the protein expression of NLRP3 and caspase-1 in muscle samples of DM/PM patients were also significantly elevated compared to that in the muscles of the controls.Conclusions:Our findings demonstrate that the NLRP3 inflammasome is implicated in the pathogenesis of DM/PM.High NLRP3 expression led to elevated levels of IL-l13 and IL-18 and could be one of the factors promoting disease progress.
基金This work was supported by the grant from the National Natural Science Foundation of China （No. 81201267）.
文摘Background： Cryopyrin-associated periodic syndrome （CAPS） is a group of rare, heterogeneous autoinflammatory disease characterized by interleukin （IL）-1β-mediated systemic inflammation and clinical symptoms involving skin, joints, central nervous system, and eyes. It encompasses a spectrum of three clinically overlapping autoinflammatory syndromes including familial cold autoinflammatory syndrome, Muckle-Wells syndrome （MWS）, and neonatal-onset multisystem inflammatory disease. CAPS is associated with gain-of-function missense mutations in NOD-like receptor family pyrin domain-containing protein 3 （NLRP3）, the gene encoding NLRP3. Moreover, most mutations leading to MWS occurred in exon 3 ofNLRP3 gene. Here, we reported a novel mutation occurred in exon 1 ofNLRP3 gene in an MWS patient and attempted to explore the pathogenic mechanism. Methods： Genetic sequence analysis of NLRP3 was performed in an MWS patient who presented with periodic lever, arthralgia, and multiform skin lesions. NLRP3 was also analyzed in this patient＇s parents and 50 healthy individuals. Clinical examinations including X-ray examination, skin biopsy, bone marrow aspiration smear, and blood test of C-reactive protein （CRP）, erythrocyte sedimentation rate （ESR）, serum levels oflL-1β, immunoglobulin E （lgE）, antineutrophil cytoplasmic antibodies, antinuclear antibodies, and extractable nuclear antigen were also analyzed. The protein structure of mutant NLRP3 inflammasome was calculated by SWISS-MODEL software. Proteins of wild type and mutant components ofNLRP3 inflammasome were expressed and purified, and the interaction abilities between these proteins were tested by surface plasmon resonance （SPR） assay. Results： X-ray examination showed no abnormality in the patient＇s knees. Laboratory tests indicated an elevation of CRP （233.24 nag/L） and ESR （67 mm/h） when the patient had fever. Serum IL-1β increased to 24.37 pg/ml, and serum lgE was higher than 2500.00 IU/ml. Other blood tests were normal. Bone marrow aspiration smear was normal. A novel point mutation c.92A〉T in exon 1 of NLRP3 gene was identified, which caused a p.D31V mutation in pyrin domain （PYD） of NLRP3. SPR assay showed that this point mutation may strengthen the interaction between the PYD of NLRP3 and the PYD of the apoptosis-associated speck-like protein. The mutation c.92A〉T in exon 1 of the NLRP3 gene was not lbund in the patient＇s parents and 50 healthy individuals. Conclusions： The rnutation c.92A〉T in exon 1 of the NLRP3 gene is a novel mutation associated with MWS. The p.D31V mutation might promote the activation ofNLRP3 inflammasome and induce MWS in this patient.
基金Supported by the National Natural Science Foundation of China(No.81960751,81660705)Guangxi Young and Middle-aged Teachers’Research Ability Improvement Project(No.2020KY59009)+2 种基金Guangxi Zhuangyao Pharmaceutical Key Laboratory(No.GXZYZZ2019-1,GXZYZZ2020-07)Guangxi Natural Science Foundation Youth Project(No.2020GXNSFBA297094)Guangxi University of Traditional Chinese Medicine School-level Project Youth Fund(No.2020QN006)。
文摘Objective:To investigate the effect of curcumol on NOD-like receptor thermoprotein domain 3(NLRP3)inflammasomes,and analyze the mechanism underlying curcumol against liver fibrosis.Methods:Thirty Kunming mice were divided into a control group,a model group and a curcumol group according to a random number table,10 mice in each group.Mice were intraperitoneally injected with 40% carbon tetrachloride(CCl4:peanut oil,2:3 preparation)at 5 m L/kg for 6 weeks,twice a week,for developing a liver fibrosis model.The mice in the control group were given the same amount of peanut oil,twice a week for 6 weeks.The mice in the curcumol group were given curcumol(30 m L/kg)intragastrically,and the mice in the model and control groups were given the same amount of normal saline,once a day for 6 weeks.Changes in liver structure were observed by hematoxylin and eosin(HE)and Masson staining.Liver function,liver fiber indices,and the expression of interleukin(IL)-10 and tumor necrosis factor-α(TNF-α)levels were determined by automatic biochemical analyzer and enzyme linked immunosorbent assay kit.Immunoblotting and reverse transcription-quantitative PCR(RT-qPCR)were performed to detect the expression of NLRP3 inflammasome-related molecules,TGF-β and collagen.Results:HE and Masson staining results showed that the hepatocytes of the model group were arranged irregularly with pseudo-lobular structure and a large amount of collagen deposition.The mice in the curcumol group had a significant decrease in liver function and liver fibers indices compared with the model group(P<0.05);RT-qPCR and Western blot results reveal that,in the curcumol group,the mRNA and protein expression levels of NLRP3,IL-1β,Caspase 1 and gasdermin D decreased significantly compared with the model group(P<0.05);immunohistochemical results showed that in the curcumol group,the protein expression levels of NLRP3 and IL-1β decreased significantly compared with the model group(P<0.05).Conclusion:A potential anti-liver fibrosis mechanism of curcumol may be associated with the inhibition of NLRP3 inflammasomes and decreasing the downstream inflammatory response.
基金supported by the State Key Program of National Natural Science of China (81930110)Military Logistics Research Project on Health Special Project (23BJZ33)the Key Project at Central Government Level: The ability establishment of sustainable use for valuable Chinese medicine resources (2060302)。
文摘Background: Idiosyncratic drug-induced liver injury(IDILI) is a serious side effect of drugs, Epimedii Folium(EF) is unequivocally implicated in idiosyncratic liver injury onset, potentially due to its ability to perturb the NOD-like receptor family pyrin domain containing 3(NLRP3) inflammasome. Fructus Ligustri Lucidi(FLL), a frequently used medicinal combination with EF, has not yet been investigated for its ability to ameliorate EF-associated hepatotoxicity. Aims and Objectives: Study on the mechanism of compatibility of FLL to alleviate liver injury caused by EF. Materials and Methods: Western blot was used to determine the expression of related proteins, ELISA was used to detect the secretion of related inflammatory factors IL-1β, IL-18, IL-6 and TNF-α, liver injury indexes were detected and liver pathological tissue staining was used to evaluate the liver injury. Results: Our results demonstrated that EF exerted a particular augmenting effect on the stimulation of the NLRP3 inflammasome mediated by nigericin or ATP, whereas FLL suppressed the NLRP3 inflammasome stimulation. Furthermore, an equal EF to FLL ratio significantly reduced the stimulatory effects of EF. Moreover, EF has the potential to induce hepatic injury and augment pro-inflammatory cytokine synthesis in rats subjected to LPS. However, when combined with FLL, the detrimental effects of EF were mitigated. Conclusions: FLL possesses the capacity to attenuate EF-associated hepatotoxicity by suppressing EF-triggered NLRP3 inflammasome activation. Thus, FLL holds promise for improving the clinical safety profile of EF, shedding light on the potential of compatibility and detoxification theories in traditional Chinese medicine.
文摘目的探讨星状神经节阻滞(SGB)通过长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)-NOD样受体热蛋白结构域相关蛋白3(NLRP3)轴在体外脑缺血再灌注模型中对炎症反应和自噬溶酶体形成的调节作用。方法培养大鼠海马神经元细胞系H19-7,并将细胞分为8组:(i)正常对照组:正常培养的神经元细胞;(ii)氧-糖剥夺/复氧(OGD/R)组:采用氧-糖剥夺/复氧法模拟脑缺血再灌注损伤;(iii)OGD/R+SGB组:OGD/R联合麻醉药0.5%布比卡因用于体外模拟SGB;(iv)OGD/R+SGB+TUG1过表达阴性对照组:OGD/R联合布比卡因并联合TUG1过表达阴性对照质粒转染细胞;(v)OGD/R+SGB+TUG1过表达组:OGD/R联合布比卡因并联合TUG1过表达质粒转染细胞;(vi)OGD/R+SGB+TUG1过表达+MCC950组:OGD/R联合布比卡因、TUG1过表达质粒转染及NLRP3抑制剂MCC950处理细胞;(vii)OGD/R+TUG1过表达组:OGD/R联合TUG1过表达质粒转染细胞;(viii)OGD/R+MCC950组:OGD/R联合NLRP3抑制剂MCC950处理细胞。进一步通过实时定量PCR(Quantitative Real Time PCR,qRTPCR)实验检测细胞中lncRNATUG1的表达;利用Western blot法检测细胞中NLRP3、微管相关蛋白1轻链3(LC3)-I、LC3-II、自噬相关基因5(Atg5)、苄氯素1(beclin1)、自噬接头蛋白(p62)、溶酶体相关膜蛋白1(LAMP1)的表达水平;利用透射电镜(TEM)检测自噬溶酶体的数量;并用酶联免疫吸附测定(ELISA)法检测细胞培养上清中白细胞介素(IL)-1β、IL-6、IL-18、肿瘤坏死因子(TNF)-α的含量。结果与正常对照组相比,OGD/R组lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05)。与OGD/R组相比,OGD/R+SGB组的上述指标均显著下调(P<0.05)。与OGD/R+SGB+TUG1过表达阴性对照组相比,OGD/R+SGB+TUG1过表达组的lncRNA TUG1、NLRP3、Atg5、beclin1、p62、LAMP1的表达水平以及LC3-II/I比值均显著上调(^(均)P<0.05),自噬溶酶体数量增加(P<0.05),IL-1β、IL-6、IL-18、TNF-α的含量显著升高(^(均)P<0.05),然而,加入NLRP3的抑制剂MCC950后,除lncRNA TUG1外其余指标均显著下调(^(均)P<0.05)。另外,与OGD/R组比,OGD/R+TUG1过表达组的上述指标进一步上调(^(均)P<0.05)。与OGD/R组比,OGD/R+MCC950组则抑制了除lncRNA TUG1外的其余指标(^(均)P<0.05)。结论星状神经节阻滞通过调节lncRNA TUG1-NLRP3轴有效减轻体外脑缺血再灌注损伤引起的炎症反应和自噬溶酶体形成,提示其可能作为治疗缺血性脑损伤的潜在策略。
基金National Natural Science Foundation of China,Grant/Award Number:82260007Jilin Province Health Commission,Grant/Award Number:2024A062+1 种基金Jilin Provincial Department of Education,Grant/Award Number:JJKH20240698KJJilin Province Science and Technology Department,Grant/Award Number:20240404025ZP and 20240602100RC。
文摘Background:This study investigated the role of polydatin in regulating macrophage-epithelial cell(EC)interactions during asthma.An asthma model was induced in BALB/c mice using ovalbumin(20μg).Methods:The therapeutic effects of polydatin(20 and 40 mg/kg)were evaluated in this asthmatic mouse model.To assess the underlying mechanisms,Bronchial Epithelium Adenovirus 12-SV402B(BEAS-2B)cells were cocultured with Tohoku Hospital for Pediatrics-1(THP-1)macrophages,in which toll-like receptor 4(TLR4)was either overexpressed or knocked down,and subsequently stimulated with lipopoly-saccharide(LPS)and ATP.THP-1 cells underwent a 1-h pretreatment with polydatin(50 and 100μmol/L),Class Lipid Inhibitor-095(CLI-095,TLR4 inhibitor,1μg/mL),or A438079(P2X7R antagonist,10μmol/L)prior to LPS/ATP challenge.Results:Findings from Western blotting,enzyme-linked immunosorbent assay,flow cytometry,real-time polymerase chain reaction,and immunofluorescence assays demonstrated that modulating TLR4 expression significantly altered interleukin-1β(IL-1β)secretion from THP-1 macrophages and mitochondrial reactive oxygen species(mtROS)production in BEAS-2B ECs.In the mouse asthma model,polydatin significantly alleviated airway inflammation,oxidative stress,and apoptosis,likely by interfering with TLR4/P2X7R-mediated signaling and suppressing the activation of the NOD-like receptor protein inflammasome.Additionally,polydatin significantly reduced IL-1βand IL-18 levels and inhibited the infiltration of macrophages and eosinophils.Correspondingly,polydatin significantly attenuated TLR4/P2X7R signaling in THP-1 cells stimulated with ATP and LPS,thereby reducing IL-1βand IL-18 secretion,calcium influx,mtROS production,and apoptosis in BEAS-2B ECs.Conclusions:Polydatin is a promising therapeutic candidate for asthma,possibly by targeting macrophage-epithelium cross-talk via the TLR4/P2X7R axis.Future formulations as capsules or sprays may effectively alleviate airway inflammation and remodeling.
