Inhibition of macrophage signaling pathway by bone marrow mesenchymal stem cells (BMSC) and alleviation of pulmonary alveolitis in mice exposed to silica dust / 上海预防医学

Objective To study the mechanism of bone marrow mesenchymal stem cell (BMSC)-mediated alleviation of pulmonary alveolitis in mice exposed to silica dust. Methods Thirty mice were randomly divided into 3 groups:control group, and two silica groups with or without BMSCs transplantation.Through the tracheal tube clearance, mice in control group received a single injection 20.0 μL of 0.90% sodium chloride solution by one time.Mice from in silica group and silica/BMSCs transplantation group first received a single injection of 20.0 μL silica dust suspension (mass concentration 250 g/L); followed by either 500.0 μL of 0.90% sodium chloride solution or by 500.0 μL of BMSCs suspension (cell density 1×109/L) through tail vein infusion 6 hours later.Mice were euthanized on the 3th day of the experiments.The levels of NALP3 inflammasome in lungs was determined by Western blot.Transwell system was used for co-culture of BMDM (in upper-chamber) and BMSC (in lower-chamber) co-culture.The level of cytokines IL-1β in BMDM cultural supernatant was detected by enzyme linked immunosorbent assay after stimulated by SiO2 stimulation.The levels of NALP3 inflammasome of in BMDM was determined by Western blot. Results The levels of pro-IL-1β, IL-1β, pro-caspase-1, caspase-1 in lungs of silica/BMSCs transplantation group were lower than that in silica group (P < 0.01).In the experiment in vitro, the concentrations of IL-1β in SiO2 exposed BMSC/BMDM co-culture group were lower than the SiO2 exposure only groups (P < 0.05).Meanwhile, the levels of pro-IL-1β, IL-1β, pro-caspase-1, caspase-1 in BMDM was lower than that in silica group (P < 0.01).The level of these proteins didn′t change while when the cell-free supernatant of BMSC culture was directly added. Conclusion The BMSC could inhibit NALP3 inflammasome of macrophages stimulated by SiO2.

Mechanism of Anti-chronic Alcoholic Liver Injury in Rats of Tibetan Medicine Lagotis brachystachys Extracts by TLR2/MyD88/NF-κB and NALP3 Signaling Pathway / 中国实验方剂学杂志

Objective::To established the model of chronic alcoholic liver injury in rats by long-term(8 weeks) alcoholic gavage, to study the effects of Tibetan medicine Lagotis brachystachys extracts on Toll-like receptor(TLR)2/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B (NF-κB)and NOD like receptor protein 3(NALP3) signaling pathways and study preliminary the mechanism of action of chronic alcoholic liver injury. Method::Sixty male Sprague-Dawley rats were randomly divided into normal group, model group, bifendate positive drug group (0.1 g·kg-1) and L. brachystachys low, medium and high-dose groups (0.5, 1, 2 g·kg-1), the corresponding drugs were given at 10 mL·kg-1 in each morning, and the 56 degree Liquor was administered by the afternoon gradient alcoholic gavage method.After 8 weeks, the levels of serum aspartate transaminase (AST), serum alanineaminotransfease(ALT), serum total cholesterol(TC), triglyceride(TG), interleukin-1β(IL-1β), and the liver levels of L-glutathione(GSH)were measured. The expression of TLR2, MyD88, NF-κB and NALP3 protein in liver were detected by Western blot.Hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue. Result::Compared with normal group, the serum levels of AST, ALT, TC, TG and IL-1β in model group were significantly increased (P<0.05, P<0.01). Compared with model group, the serum AST, ALT, TC, TG and IL-1β levels were decreased in the various doses of L. brachystachys, and the high dose group was particularly effective (P<0.05, P<0.01). Compared with normal group, the GSH level in the liver homogenate of model group decreased significantly, and the difference was not statistically significant. The levels of TLR2, MyD88, NF-κB and NALP3 in the liver tissue of model group were significantly increased (P<0.05, P<0.01). The GSH levels in the liver and the protein expression of TLR2, MyD88, NF-κB and NALP3 were decreased in L. brachystachys group (P<0.05, P<0.01). The liver pathological section showed that L. brachystachys can improve the pathological changes of rat liver tissue. Conclusion::L. brachystachys can protect liver from alcohol-induced chronic liver injury in rats. The mechanism was related to TLR2/MyD88/NF-κB and NALP3 signaling pathway.

Anti-inflammatory Effect of Total Saponin Fraction from Dioscorea nipponica Makino on Gouty Arthritis and Its Influence on NALP3 Inflammasome / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of Chinese herbal medicine Dioscorea nipponica for the treatment of monosodium urate crystals-induced gouty arthritis (GA) in rats.@*METHODS@#Sixty male Wistar rats were divided into 6 groups: normal, model, indomethacin and three total saponin (900, 300 and 100 mg/kg) groups. The liver, kidney and serum levels of lysosomal enzymes, antioxidant capacities, and inflammatory factors were measured. In addition, the mRNA and protein levels of the NALP3 inflammasome components in the mononuclear cells of rats' peripheral blood were analyzed using real-time polymerase chain reaction and Western blotting methods, respectively.@*RESULTS@#Total saponins groups could reduce the activities of β-galactosidase, β-N acetyl glucosamine enzyme, β-glucuronidase, acid phosphatase, and malonaldehyde as well as the contents of TNF-α, IL-1β and IL-8 (all P<0.05). They could also increase the activities of glutathione peroxidase and total superoxide dismutase (both P<0.05). Further studies showed that total saponins groups of high, middle and low doses could all increase the mRNA and protein levels of caspase-1, adapter apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) and NALP3 in the mononuclear cells of peripheral blood (all P<0.05).@*CONCLUSION@#Dioscorea nipponica may treat GA by regulating lysosomal enzymes, antioxidant capacities and the NALP3 inflammasome.

The function and mechanism of NALP3 inflammasome in interstitial cystitis/bladder pain syndrome / 中华泌尿外科杂志

Objective To acknowledge the NALP3 inflammasome expression and significance in the interstitial cystitis/bladder pain syndrome (IC/PBS).Methods The urine of 16 IC/BPS patients and 16 normal persons was collected to measure the IL-1β content by ELISA.Bladder tissue of 16 IC/BPS patients and para-carcinoma tissue of 16 bladder cancer patients were collected.And the levels of NALP3,caspase1 and IL-1β were detected by Western Blot.60 female rats were randomly divided into control group(bladder was infused with 0.5 ml saline),hyaluronidase group [bladder was infused with 0.5 ml hyaluronidase (4 mg/ml)],NALP3 antagonist group [bladder was infused with 0.5 ml hyaluronidase (4 mg/ml) and Glyburide(10 mg/kg)] and mucosal protectant group [bladder was infused with 0.5 ml hyaluronidase (4 mg/ml) and sodium hyaluronate(0.8 mg/ml)] to carried out the animal experiment,and 15 rats in each group.The models were created by long-term (1 month) intermittent intravesical hyaluronidase infusion.Voiding patterns were investigated by cystometry.Toluidine blue staining was used to detected mast cell’s changes.The levels of NALP3,caspase-1 and IL-1β were determined by Western Blot,HE staining was to detect tissue inflammation of the bladder,and the severity of pain was examined by Von-frey brush by using the strength of 0.07、0.4、1.0 g.The comparison between the chemotaxis of 200 ng,400 ng IL-1β and 200ng SCF IL-1β to mast cells was checked by Transwell experiment.Results The expressions of IL-1β in IC/PBS patients was increased in IC/PBS group than normal control group [(381 ± 112) μg/L vs.(98 ± 40) μg/L,P ＜0.01].The expressions of NALP3,Caspase-1 and IL-lβ had increased in the IC/PBS group than normal group(0.22 ±0.08 vs.0.11 ±0.02,0.25 ±0.03 vs.0.10 ±0.01,0.19 ±0.04 vs.0.11 ± 0.02,P ＜ 0.05)by Western Blot.In the IC/PBS rats,compared with the control group,the intercontraction intervals [(120.0 ± 15.6) s vs.(447.3 ± 24.6) s] and bladder capacity [(0.34 ± 0.02) ml vs.(1.33 ± 0.04) ml] of the model group were significantly decreased (both P ＜ 0.05).In mucosal protectant group and NALP3 antagonist group,the intercontraction intervals [(323 ± 16.3)s,(280 ± 12.5)s] and bladder capacity [(1.14 ± 0.05) ml,(0.84 ± 0.04) ml] were increased compared with control group (P ＜ 0.05).The amount of mast cell in model group were significantly increased than control group (3.4 ±0.8 vs.0.4 ± 0.2,P ＜ 0.05) while in mucosal protectant group (1.8 ± 0.5) and NALP3 antagonist group (1.5 ± 0.7) were decreased compared with control group (P ＜ 0.05).The protein levels in modle group of NALP3 (5.91 ±0.33 vs.1.00 ±0.12),caspase-1 (6.75 ±0.42 vs.1.00 ±0.22) and IL-1β(7.12 ±0.45 vs.1.00 ± 0.18)were increased than control group.In mucosal protectant group and NALP3 antagonist group,theNALP3 (2.921 ±0.21,2.07±0.18),caspase-1 (3.28 ±0.31,2.25 ±0.19) and IL-1β(3.33± 0.41,1.98 ±0.21) were decreased compared with control group.VonFrey pain score in model group were significantly increased than control group(0.07 g:7.5 ± 1.8 vs.2.1 ± 0.5,0.4 g:9.2 ± 1.9 vs.5.2 ± 1.1,1.0g:15.4±3.8 vs.6.8±1.5,P＜0.05) and VonFrey pain score(0.07 g:2.4±0.3,2.8± 0.7;0.4 g:5.2 ±0.4,6.5 ±1.3;1.0 g:6.4 ±0.8,7.3 ±1.1;P＜0.05) in NALP3 antagonist group were significantly decreased.In vitro,Transwell experimental results showed that 400 ng IL-1β of mast cell chemotaxis is similar with that of the 200 ng SCF (3 800 ±400 vs.4 800 ±500,P ＞0.05).Conclusions The levels of NALP3/Caspase-1/IL-1β in the urine of patients with IC/PBS were significantly higher than those in normal control group.NALP3 is activated in chronic cystitis rat model,and related to pain and frequent urination.This may be related to the down-regulation of expression of NALP3,caspase-1,IL-1β,and other inflammatory mediators,and blocking the chemotactic effects of IL-1 β on mast cells.

Role and significance of Febuxostat on NALP3 inflammasome in chronic gouty arthritis / 实用医学杂志

Objective To study the effect of Febuxostat on NALP3 inflammasome in chronic gouty arthri-tis. Methods A total of 89 patients with chronic gouty arthritis and 50 healthy cases were enrolled in this study and 89 patients were divided into Benzbromarone group,Allopurinol group,Febuxostat group and placebo group. The expression of NALP3 inflammasome mRNA and the protein were detected by RT-PCR and Western blot. Re-sults The levels of Uric Acid,NALP3,ASC and caspase-1 mRNA in chronic gouty arthritis patients were higher than those in healthy cases before treatment(#P = 0.000);the level of NALP3 in benzbromarone and allopurinol group had no change after treatment(*P<0.05).The levels of NALP3 mRNA and caspase-1 mRNA in Febuxostat group were lower but the level of ASC mRNA was higher than those in other groups after treatment(*P < 0.05). Conclusions NALP3 inflammatory may be associated with chronic gouty arthritis. Febuxostat can effectively re-duce the level of Uric Acid,and affect the function of NALP3 inflammasome.

Effect of total saponin of Dioscorea on NALP3 inflammasome signaling pathway with acute gouty in rats / 中国药理学通报

Aim Toinvestigatetheeffectsoftotalsap-onin of Dioscorea (TSD)on rats with monosodium u-rate crystal-induced acute gouty arthritis (AGA)and mechanisms.Methods Totally72Wistarratswere randomly devided into six groups,Each group was giv-en corresponding drug before,then rat acute gouty ar-thritis model was made by injection of monosodium u-rate in the ankle joint cavity.The gait,articular swell-ing degree and physiological changes of rats were ob-served.The concentration of TNF-α,IL-1β,IL-18 in serum were detected by ELISA.The levels of pro-IL-1β, NALP3, ASC, pro-caspase-1, and cleaved caspase-1 were detected by Western blot.Results AllTSDgroupsandcolchicinesignificantlychangedthe gait of rats and TSD high and middle groups signifi-cantly reduced joint swelling and diminished the patho-logical changes.The levels of TNF-α,IL-1β,IL-18 in serum were significantly decreased,and the levels of pro-IL-1β,NALP3,ASC,pro-caspase-1 and cleaved caspase-1 were apparently reduced in TSD high and middlegroups.Conclusion TSDpossessesanti-goutfunction and the mechanism may be related to sup-pressing the NALP3 inflammasome activation and in-hibiting the cytokine production.

El inflamasoma: mecanismos de activación / Inflammasome: activation mechanisms

La inflamación es una respuesta biológica rápida del sistema inmune en tejidos vasculares, dirigida a eliminar estímulos capaces de producir daño y a iniciar la curación y la reparación. Los complejos macromoleculares denominados inflamasomas están constituidos por un receptor NOD (NLR), un receptor de AIM2 (ausente en melanoma 2) el ALR, la proteína tipo punto asociada a apoptosis (ASC) y la procaspasa-1, los cuales pueden ser activados por variación en la concentración iónica y de ATP intracelular y extracelular, por desestabilización del fagolisosoma, por internalización de cristales insolubles y por mecanismos de oxidoreducción, lo cual permitirá la activación de la plataforma molecular y el consiguiente procesamiento de las prointerleuquinas inflamatorias a sus formas activas. En la actualidad existen dos nodos de señalización utilizados por los inflamasomas: canónica y no canónica para generar respuestas efectoras. Datos recientes vinculan al inflamasoma NLRP3, la IL-1b y a la IL-18, en el desarrollo y evolución de enfermedades tales como: ateroesclerosis, diabetes tipo II, hiperhomocisteinemia, gota, malaria e hipertensión arterial e identificaron esta cascada, como un blanco quimioterapéutico ideal para la prevención de estas patologías. En esta revisión se discutirán los mecanismos de activación y regulación del inflamasoma que estimulan, modulan y resuelven los procesos inflamatorios.

Inflammation is a rapid biologic response of the immune system in vascular tissues, directed to eliminate stimuli capable of causing damage and begin the process of repair. The macromolecular complexes known as “inflammasomes” are formed by a receptor, either NOD (NLR) or ALR, the receptor absent in melanoma 2 (AIM2). In addition, the inflammasome is formed by the speck-like protein associated to apoptosis (ASC) and procaspase-1, that may be activated by variations in the ionic and intracellular and extracellular ATP concentrations; and the loss of stabilization of the fagolisosomme by internalization of insoluble crystals and redox mechanisms. As a result, there is activation of the molecular platform and the processing of inflammatory prointerleukins to their active forms. There are two modalities of activation of the inflammasome: canonical and non-canonical, both capable of generating effector responses. Recent data associate NLRP 3, IL-1b and IL-18 in the pathogenesis of a variety of diseases, including atherosclerosis, type II diabetes, hyperhomocysteinemia, gout, malaria and hypertension. The inflammasome cascade is emerging as a new chemotherapeutic target in these diseases. In this review we shall discuss the mechanisms of activation and regulation of the inflammasome that stimulate, modulate and resolve inflammation.

pORF5 plasmid protein of Chlamydia trachomatis induces IL-1βand IL-18 production through NALP3 inflammasome activation / 中国免疫学杂志

Objective:To investigate whether pORF5 plasmid protein of Chlamydia trachomatis(Ct) induces 1L-1βand 1L-18 production in THP-1 cells,and its potential molecular mechanism.Methods:pORF5 plasmid protein was used to stimulate THP-1 cells at different concentrations(0,3,6,12,24,36 μg/ml),then the inflammatory cytokines IL-18 and IL-1βwere detected by ELISA at the time of 0,8,16,24,36 h;The mRNA expression of NALP3 inflammasome were detected by Realtime-PCR,and Caspase-1 activity was determined by Western blot analysis.THP-1 cells were transfected with siRNA targeting NALP3 and ASC gene for 24 h or pretreated with Caspase-1 inhibitor(Z-YVAD-FMK) for 30 min,and subsequently stimulated with pORF5(24 μg/ml) for 24 h,then secretion of IL-1βand IL-18 were analyzed by ELISA.Results: The pORF5 plasmid protein induced THP-1 cells to secrete IL-1βand IL-18 by dose-and time-dependent manners,production of IL-1βand IL-18 reached their peaks(491 pg/ml and 186 pg/ml) at concentration of 24 μg/ml,and the peak amount of IL-1βand IL-18 occurred at 24 h and 16 h post-stimulation respectively.pORF5 plasmid protein in-creased mRNA expression of NALP3 inflammasome and activated Caspase-1 in THP-1 cells.NALP3 siRNA,ASC siRNA and Z-YVAD-FMK reduced pORF5-induced IL-1βand IL-18 production when compared with control groups(P<0.05).Conclusion:pORF5 plasmid protein could induce THP-1 cells to produce IL-1βand IL-18 through NALP3 inflammasome activation,which may play an important role in the pathogenesis in Ct infection.

Polyionic vaccine adjuvants: another look at aluminum salts and polyelectrolytes

Adjuvants improve the adaptive immune response to a vaccine antigen by modulating innate immunity or facilitating transport and presentation. The selection of an appropriate adjuvant has become vital as new vaccines trend toward narrower composition, expanded application, and improved safety. Functionally, adjuvants act directly or indirectly on antigen presenting cells (APCs) including dendritic cells (DCs) and are perceived as having molecular patterns associated either with pathogen invasion or endogenous cell damage (known as pathogen associated molecular patterns [PAMPs] and damage associated molecular patterns [DAMPs]), thereby initiating sensing and response pathways. PAMP-type adjuvants are ligands for toll-like receptors (TLRs) and can directly affect DCs to alter the strength, potency, speed, duration, bias, breadth, and scope of adaptive immunity. DAMP-type adjuvants signal via proinflammatory pathways and promote immune cell infiltration, antigen presentation, and effector cell maturation. This class of adjuvants includes mineral salts, oil emulsions, nanoparticles, and polyelectrolytes and comprises colloids and molecular assemblies exhibiting complex, heterogeneous structures. Today innovation in adjuvant technology is driven by rapidly expanding knowledge in immunology, cross-fertilization from other areas including systems biology and materials sciences, and regulatory requirements for quality, safety, efficacy and understanding as part of the vaccine product. Standardizations will aid efforts to better define and compare the structure, function and safety of adjuvants. This article briefly surveys the genesis of adjuvant technology and then re-examines polyionic macromolecules and polyelectrolyte materials, adjuvants currently not known to employ TLR. Specific updates are provided for aluminum-based formulations and polyelectrolytes as examples of improvements to the oldest and emerging classes of vaccine adjuvants in use.

Expression change of NALP3/ASC/caspase-1 in renal tissues of FSGS / 重庆医学

Objective To investigate the expression change of renal NLR family pyrin domain containing-3 protein(NALP3) in-flammasome in the nephrotic syndrome(NS) patients with focal segmental glomerulosclerosis(FSGS) and its relation with the tubu-lointerstitial pathogenic injury degree ,expression of inflammatory factors and clinical biochemical indexes .Methods Immunohisto-chemistry was used to detect the expressions of NALP3/ASC/caspase-1 and their downstream effector molecule IL-1β,IL-18 in re-nal tubular epithelial cells .The tubulointerstitial injury score and the activated macrophages F4/80 in renal interstitium of the FSGS patients and NS patiens were evaluated .The serum creatinine ,urea ,total protein ,albumin ,24 h urine protein and estimated glomer-ular filtration rate(eGFR) were observed .The correlation of tubulointerstitial injury with NALP3/ASC/caspase-1 ,IL-1β,IL-18 were respectively analyzed .Results The expression of NALP3/ASC/caspase-1 ,IL-1β,IL-18 in the renal tissue of the FSGS pa-tients was significantly increased compared with that in the control group (P<0 .01) .NALP3/ASC/capspase-1 expression was pos-itively correlated with the expression of IL-1β,IL-18(P< 0 .01) .NALP3/ASC/caspase-1 ,IL-1β,IL-18 expression was positively correlated with renal tubulointerstitial injury and the F4/80 expression intensity(P<0 .01) .NALP3/ASC/caspase-1 ,IL-1β,IL-18 was significantly positively correlated with 24 h urine protein and Scr ,and negatively correlated with the eGFR (P<0 .05) ,but had no obvious correlation with plasma urea ,plasma total protein and albumin concentrations .Conclusion The NALP3 inflammasome might participate in the pathogenic mechanism of FSGS through the activation of its downstream inflammatory factor of IL-1β,IL-18 ,the more higher its expression degree ,the more severe the renal tissue injury ,whether which could be served as the warning in-dex needs the further clinical verification .

Protective mechanism of NALP3-siRNA on rat renal tubular epithelial cells from hypoxia/reoxygenation injury / 中华肾脏病杂志

Objective To explore the mechanism of protecting cells from hypoxia/ reoxygenation (H/R) injury by constructing specific small interference RNA (siRNA) to inhibit NALP3 expression in rat renal tubular epithelial cells (NRK-52E).Methods (1) To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 1 h,2 h,4 h,8 h,16 h and 24 h.The activity of lactae dehydrogenase (LDH) in the culture medium,cell count and cell viability,the expression of NALP3 were determined by biochemical method,trypan blue exclusion and Western blotting.(2) The siRNA was transfected into NRK-52E.The irrespective siRNA transfected group wasused as control.NALP3 expression was examined by Western blotting.(3) The cells were divided into 4 groups:control group,H/R group,irrespective siRNA transfected group and NALP3-siRNA transfected group.To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 4 h.And the expression of NALP3 was determined by Western blotting.(4)Cellular apoptosis was examined by Annexin V/PI staining and flow cytometry.NF-κB DNA binding activity,IκB-α,Bcl-2 and Bax expression were examined by EMSA and Western blotting.Results (1)Compared with the control group,the activity of LDH significantly increased,cell count and cell viability significantly decreased (all P＜0.05).The expression of NALP3 significantly increased and peaked at 4 h after H/R.(2)The specific siRNA could efficiently inhibit NALP3 expression in NRK-52E.Compared with the irrespective siRNA transfected group,the protein expression of NALP3 was significantly down-regulated in NALP3 siRNA transfected group (P＜0.05).(3)After hypoxia 1 h and reoxygenation 4 h,the activity of LDH and the expression of NALP3 increased.Compared with the irrespective siRNA transfected group,LDH concentration in media and the expression of NALP3 significantly decreased in NALP3-siRNA transfected group.(4)After hypoxia 1 h and reoxygenation 4 h,NF-κB DNA binding activity was increased,IκB-α phosphorylation and degradation,Bcl-2 and Bax were significantly up-regulated.However,compared with the irrespective siRNA transfected group,NF -κB DNA binding activity,IκB-α degradation and Bax/Bcl-2 were significantly decreased (P＜0.05) in NALP3-siRNA transfected group.At the same time,the ratio of apoptosis was significantly increased in three groups than that in control.Compared with the irrespective siRNA transfected group,the ratio of apoptosis in NALP3-siRNA transfected group was significantly decreased (P＜0.05).Conclusions H/R induces the expression of NALP3 in NRK-52E.The synthesized siRNA can inhibit the expression of NALP3 and protect NRK-52E from hypoxia/reoxygenation injury.The mechanism may be via inhibiting the activation of NF-κB,modulating expression of Bcl-2 and Bax,as well as decreasing cell apoptosis.

Expression of NALP3 in the Spleen of Mice with Portal Hypertension / 华中科技大学学报（医学）（英德文版）

This study examined the mRNA expression of NALP3 in the spleen of the mice with hypersplenism due to portal hypertension(PH).The mouse hypersplenism models were established by oral administration of tetrachloromethane(2 mL/kg/week for 12 weeks by oral gavage).All the mice were randomly divided into a control group and an experimental group.The blood routine test was conducted,spleen index was calculated and spleen was histologically examined.Portal vein sera were taken for detection of the level of uric acid.The mRNA expressions of NALP3 and IL-1β in the spleen were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).The results showed that the platelet count was significantly lower in the experimental group[(674±102)× 109/L]than in the control group[(1307± 181)× 109/L](P<0.05),while the spleen index was significantly higher[(9.83±1.36)μg/g]in the experimental group than in the control group[(4.11±0.47)μg/g](P<0.05).The histopathological changes of spleen followed the pattern of congestive splenomegaly.No significant difference was found in the uric acid level in the portal vein between the control group and the experiment group.The mRNA expressions of NALP3 and IL-1β were up-regulated significantly in the spleen in the experimental group as compared with those in the control group(P<0.05).It was concluded that NALP3 and IL-1β may play important roles in the pathogenesis of hypersplenism.