Jeju seaweeds suppress lipopolysaccharide-stimulated proinflammatory response in RAW 264.7 murine macrophages

Objective:To investigate the anti-inflammatory effects of Jeju seaweeds on macrophage RAW264.7 cells under lipopolysaccharide(LPS)stimulation.Methods:Ethyl acetate fractions were prepared from five different types of Jeju seaweeds,Dictyopteris divaricata(D.divaricata),Dictyopteris prolifera(D.prolifefa),Prioutis cornea(P.comea,Grateloupia laceolata(G,lanceolate,and Cralcloupia filicina(G.filicina)They were screened for inhibitory effects on proinflammatory mediators and cytokines such as nitric oxide(NO),prostaglandin E,,tumor necrosis factor-a(TNF-a),and interleukin-6(11.-6).Results:Our results revealed that D.divaricata,D.prolifera,P.cornea,G.lanceolata,and G.filicina potently inhibited I.PS-stimulaled NO production(IC_(50),values were 18.0,38.36,38.43,32.81 and 37.14μg/mL,respectively).Consistent with these findings,D.divtricata,D.prolifera,P.cornea,and G.fdicina also reduced the IPS-induced and prostaglandin E,production in a concentration-dependent manner.Expectedly,they suppressed the expression of inducible NO synthase and cyclooxygenase-2 at the protein level in a dose-dependent manner in the RAW264.7 cells,as detennined by western blotting.In addition,the levels of TNF-a and IL-6,released into the medium,were also reduced by D.divaricata,D.prolifera,P.cornea,G,lanceolata,and G.fdicina in a dose-dependent manner(IC_(50)values for TNF-a were 16.11,28.21,84.27,45.52 and74.75μg/mL,respectively;IC_(50),values for IL-6 were 37.35,80.08,103.28,62.53 and 84.28μg/mL,respectively).The total phlorotannin content was measured by the Folin-Ciocalteu method and expressed as phloroglucinol equivalents.The content was 92.0μg/mg for D.divaricata,151.8μg/mg for D.prolifera,57.2μg/mg for P.cornea,53.0 pg/mg for G.lanceolata,and 40.2μg/mg for G.fdicina.Conclusions:Thus,these findings suggest that Jeju seaweed extracts have potential therapeutic applications for inflammatory responses.

Aloe barbadensis Miller peptide/polypeptide fraction alleviates inflammation through inhibition of proinflammatory cytokines and mediators in vitro and in rats with Freund’s adjuvant-induced hind paw edema

Objective:To evaluate the anti-inflammatory potential of peptide/polypeptide fraction of Aloe vera through in vitro and in vivo studies.Methods:The peptide/polypeptide fraction from Aloe vera was obtained through trichloroacetic acid precipitation.The anti-inflammatory property of the peptide/polypeptide fraction was tested by protein denaturation,membrane stabilization assays.The effect of the fraction on RAW 264.7 cell viability was examined by MTT assays.The nitric oxide level was determined through Griess reagent.TNF-αand IL-6 levels were estimated using ELISA kits.In vivo studies were carried out in male Wistar rats through injection of Freund’s adjuvant in the hind paw.Paw edema was measured through the Vernier scale and levels of alanine aminotransferase,aspartate transaminase,TNF-α,IL-6,and secretory phospholipase A2 were estimated through their respective kits after fourteen days of treatment.Graph Pad Prism6 was used for analyzing the results.Results:The peptide/polypeptide extract inhibited protein denaturation with an IC50 value of(218.9±15.6)μg/m L and stabilized the membrane of red blood cells with an IC50 value of(275.9±19.1)μg/m L.The extract showed no changes in cell morphology or cytotoxicity up to the concentration of 20μg/mL in MTT assays.The peptide/polypeptide fraction markedly reduced the levels of proinflammatory markers and mediators in both in vitro and in vivo studies.Conclusions:The results indicate that the peptide/polypeptide fraction of Aloe vera has antiinflammatory property through inhibition of inflammatory markers and mediators responsible for NF-κB and mitogen-activated protein kinase pathways.

Regulation of the expression of proinflammatory cytokines induced by SARS-CoV-2

An outbreak of a novel coronavirus was reported in Wuhan,China,in late 2019.It has spread rapidly through China and many other countries,causing a global pandemic.Since February 2020,over 28 countries/regions have reported confirmed cases.Individuals with the infection known as coronavirus disease-19(COVID-19)have similar clinical features as severe acute respiratory syndrome first encountered 17 years ago,with fever,cough,and upper airway congestion,along with high production of proinflammatory cytokines(PICs),which form a cytokine storm.PICs induced by COVID-19 include interleukin(IL)-6,IL-17,and monocyte chemoattractant protein-1.The production of cytokines is regulated by activated nuclear factor-kB and involves downstream pathways such as Janus kinase/signal transducers and activators transcription.Protein expression is also regulated by post-translational modification of chromosomal markers.Lysine residues in the peptide tails stretching out from the core of histones bind the sequence upstream of the coding portion of genomic DNA.Covalent modification,particularly methylation,activates or represses gene transcription.PICs have been reported to be induced by histone modification and stimulate exudation of hyaluronic acid,which is implicated in the occurrence of COVID-19.These findings indicate the impact of the expression of PICs on the pathogenesis and therapeutic targeting of COVID-19.

Emodin suppresses LPS-induced proinflammatory responses and nuclear factor-B activation by disruption of lipid rafts and TLR-4 recruitment in endothelial cells

Emodin [1,3,8-Trihydroxy-6-methylanthraquinone] has been reported to exhibit vascular anti-inflammatory properties.However,the relevant anti-inflammatory mechanisms are not well understood.The present study was design to explore the molecular target(s) of emodin

Changes of proinflammatory, fluid balance, vascular permeability and oxidative stress in patients with secretory otitis media

Objective: To investigate the changes of proinflammatory, fluid balance, vascular permeability and oxidative stress in patients with secretory otitis media. Methods: A total of 42 patients with secretory otitis media treated in our hospital from September 2016 to January 2018 were selected as the observation group and 42 healthy people as the control group. The levels of proinflammatory effect [including transforming growth factor beta 1 (TGF-β1), transforming growth factor-beta 2 (TGF-β2)], fluid balance [including aquaporin-1 (AQP-1) and aquaporin-4 (AQP-4)], vascular permeability [including hyaluronic acid (HA), fibronectin (Fn) and platelet activating factor (PAF)] and oxidative stress-related indexes [including malondialdehyde (MDA) and superoxide dismutase (SOD)] were observed and compared between the two groups. Results: The levels of AQP-1, AQP-4 and Fn in the observation group were significantly lower than those in the control group. The expression levels were (4.52±0.39) g/L, (23.06±7.21) g/L and (120.59±13.07) mg/L, respectively. The levels of TGF-β1, TGF-β2, HA, PAF, MDA and SOD were significantly higher than those of the control group and the expression levels were (11.99±4.23) 毺g/L, (4.22±1.66) 毺g/L, (70.54±6.99) 毺g/L, (123.83±20.58) ng/mL, (6.30±0.44) nmol/mL and (15.85±0.78) NU/mL, respectively. The difference was statistically significant. Conclusions: Inflammatory reaction occurs in patients with otitis media with secretory otitis media. It is easy to break the liquid balance in the middle ear cavity and to increase vascular permeability and oxidative stress. The relevant indicators should be strengthened in clinical practice, so as to provide evidence for early diagnosis and treatment of the disease.

The Diagnostic Value of the Investigation of the Proinflammatory and Anti-inflammatory Cytokines Levels in the Immune System of Patients with Breast Cancer

Diagnostic value of serumal levels of proinflammatory(TNF-α,IL-1β,IL-6 and IFN-γ) and anti-inflammatory(IL-4 and IL-10) cytokines of immune system is investigated at 54 mammary glands sick by a cancer.The analysis carried out has allowed to tap the raised concentration of such proinflammatory cytokines,as IL-1β,TNF-α and IL-6 in serum of peripheric blood of MGC patients.The analysis of antiinflammatory cytokines in Serum of peripheric blood of MGC patients also has taped authentically raised value of IL-4 and IL-10.The obtained data testify to presence imbalance cellular and humoral factors of the immune system which studying will serve as the important diagnostic criterion of breast cancer.

Ammonia-induced oxidative stress triggered proinflammatory response and apoptosis in pig lungs

Ammonia,a common toxic gas,is not only one of the main causes of haze,but also can enter respiratory tract and directly affect the health of humans and animals.Pig was used as an animal model for exploring the molecular mechanism and dose effect of ammonia toxicity to lung.In this study,the apoptosis of type II alveolar epithelial cells was observed in high ammonia exposure group using transmission electron microscopy.Gene and protein expression analysis using transcriptome sequencing and western blot showed that low ammonia exposure induced T-cell-involved proinflammatory response,but high ammonia exposure repressed the expression of DNA repair-related genes and affected ion transport.Moreover,high ammonia exposure significantly increased 8-hydroxy-2-deoxyguanosine (8-OHdG) level,meaning DNA oxidative damage occurred.In addition,both low and high ammonia exposure caused oxidative stress in pig lungs.Integrated analysis of transcriptome and metabolome revealed that the up-regulation of LDHB and ND2 took part in high ammonia exposure-affected pyruvate metabolism and oxidative phosphorylation progress,respectively.Inclusion,oxidative stress mediated ammonia-induced proinflammatory response and apoptosis of porcine lungs.These findings may provide new insights for understanding the ammonia toxicity to workers in livestock farms and chemical fertilizer plants.

Production of proinflammatory cytokines in the human THP-1 monocyte cell line following induction by Tp0751,a recombinant protein of Treponema pallidum

The tissue destruction characteristic of syphilis infection may be caused by inflammation due to Treponema pallidum and the ensuing immune responses to the pathogen.T.pallidum membrane proteins are thought to be potent inducers of inflammation during the early stages of infection.However,the actual membrane proteins that induce inflammatory cytokine production are not known,nor are the molecular mechanisms responsible for triggering and sustaining the inflammatory cascades.In the present study,Tp0751 recombinant protein from T.pallidum was found to induce the production of proinflammatory cytokines,including TNF-α,IL-1βand IL-6,in a THP-1 human monocyte cell line.The signal transduction pathways involved in the production of these cytokines were then further investigated.No inhibition of TNF-a,IL-1β,or IL-6 production was observed following treatment with the SAPK/JNK specific inhibitor SP600125 or with an ERK inhibitor PD98059.By contrast,anti-TLR2 mAb,anti-CD14 mAb,and the p38 inhibitor SB203580 significantly inhibited the production of all three cytokines.In addition,pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-κB,profoundly inhibited the production of these cytokines.Tp0751 treatment strongly activated NF-κB,as revealed by Western blotting.However,NF-κB translocation was significantly inhibited by treatment with PDTC.These results indicated that TLR2,CD14,MAPKs/p38,and NF-κB might be implicated in the inflammatory reaction caused by T.pallidum infection.

Characteristics of Proinflammatory Cytokines and Chemokines in Airways of Asthmatics： Relationships with Disease Severity and Infiltration of Inflammatory Cells

Background：Increased proinflammatory cytokines and chemokines might contribute to infiltration of inflammatory cells and remodeling in airways of asthma.Although these molecules may be associated with asthma,there is lack of systemic evidence showing which and how important these events are in the disease.We aimed to analyze the concentrations of these molecules in the airways and relationships with disease severity and with airway infiltration of inflammatory cells in a large cohort of asthmatics （n =70,including 37 mild and 33 moderate/severe asthmatics） compared with controls （n =30）.Methods：Meso scale discovery system and commercial ELISA kits were used to measure the concentrations ofproinflammatory cytokines interleukin （IL）-1 β;tumor necrosis factor-alpha （TNF-α）;IL-6;and IL-17 and CC and CXC chemokines CCL2,CCL4,CCL 11,CCL 13,CCL17,CCL22,and CCL26 and CXCL8,CXCL9,CXCL10,and CXCL1 1 in bronchoalveolar lavage fluid of asthmatics and controls.Results：The concentrations ofIL-1,TNF-α,IL-6,CXCL8 and CXCL 10,and CCL4,CCL 11,CCL 17,and CCL22 were significantly elevated in asthmatics compared with controls （P 〈 0.05）.The concentrations of TNF-α and CXCL8,but not others,were negatively correlated with severity of disease （lung function forced expiratory volume in 1 s） （TNF-α vs.total：r =-0.359,P =0.002 vs.moderate/severe：r =-0.541,P =0.001;CXCL8 vs.total：r =-0.327,P =0.006 vs.moderate/severe：r =-0.625,P =0.0001,respectively）.In addition,concentrations of these two molecules were also correlated with the absolute numbers of infiltrating eosinophils and neutrophils in asthmatic airways.Conclusions：Increased concentrations of TNF-α and CXCL8 are associated with pathogenesis of asthma.Targeting these molecules might provide an alternative therapeutic for this disease.

Phonopheresis Associated with Nanoparticle Gel from Phyllanthus amarus Relieves Pain by Reducing Oxidative Stress and Proinflammatory Markers in Adults with Knee Osteoarthritis

Objective: To determine the changes in serum levels of inflammatory biomarkers and antioxidant levels among the knee osteoarthritis(OA) patients after treatment with Phyllanthus amarus(PP) by nanoparticle gel phonophoresis. Methods: This study was a randomized, double-blind, placebo-control, parallel-group, clinical trial involving 30 subjects with mild-to-moderate degree of knee OA. The patients were allocated to two groups using a computer-generated random numbers, and received conventional ultrasound therapy(control group, 15 cases) and PP(treatment group, 15 cases) once daily for 10 sessions. The pain was evaluated by visual analogue scale(VAS). Serum levels of tumor necrosis factor-α(TNF-α) were determined by enzyme-linked immunosorbnent assay(ELISA). Nitric oxide(NO) was determined by modified Griess reagent. The antioxidant effects, including superoxide dismutase(SOD) and total antioxidant capacity(TAC), were also measured by ELISA assay. Results: The VAS score was significantly decreased in the treatment group compared with the control group after treatment(P<0.01). The serum concentrations of TNF-α and NO were significantly reduced in the treatment group compared with the control group(P<0.01) after treatment. However, the serum concentrations of SOD and TAC in the treatment group were significantly higher after treatment compared with the control group(P<0.01). Conclusion: PP could alleviate knee pain and significantly reduce systemic antiinflammatory effects in knee OA patients.

Drug-containing serum of Xinfeng capsules protect against H9C2 from death by enhancing miRNA-21 and inhibiting toll-like receptor 4/phosphorylated p-38(p-p38)/p-p65 signaling pathway and proinflammatory cytokines expression

OBJECTIVE: To investigate effect of drug-containing serum of Xinfeng capsules on myocardial cell growth.METHODS: Drug-containing serum of Xinfeng capsules rat models were established by intragastricly administrated Xinfeng capsules. MTT assay wasused to evaluated H9C2 cells viability. H9C2 cells were divided into normal control group, triptolide group, lipopolysaccharide(LPS) group, drug-containing serum group and mi RNA-21 inhibitor group. micro RNA-21(mi RNA-21) inhibitor was structured and transfected into H9C2 cells. Western blot and immunofluorescence assay were applied to examine toll-like receptor 4(TLR4), phosphorylated p-38(p-p38) and p-p65 expression. Quantitative real-time PCR(q RT-PCR) was used to evaluated m RNA levels of mi RNA-21. Enzyme linked immunosorbent(ELISA) was used to measure tumor necrosis factor α(TNF-α), IL-6 and IL-17 levels.RESULTS: Drug-containing serum treatment significantly increased cell viability compared to LPS treated group. q RT-PCR results indicated that mi RNA-21 levels were significantly decreased in drugcontaining serum group compared to LPS group.Early and late apoptosis in drug-containing serum group were significantly decreased compared to LPS group. Western blot and immunofluorescence assay results showed that TLR4, p-p38 and p-p65 levels in drug-containing serum group were significantly decreased compared to LPS group. ELISA findings indicated that drug-containing serum significantly decreased inflammatory cytokine levels of TNF-α, IL-6 and IL-17.CONCLUSION: Drug-containing serum of Xinfeng capsules protect against lipopolysaccharide instructed H9C2 cells from death by enhancing mi RNA-21 and inhibiting TLR4/p-p38/p-p65 signaling pathway and proinflammatory cytokines expression.

Reproductive stage associated changes in plasma fatty acid profile and proinflammatory cytokine expression in rat mammary glands

Mastitis is a common disease for mammals all around the world. Figuring out why mastitis mainly occurs around parturition may be helpful for dealing with the disease. Lipolytic activity and oxidative stress take place around parturition, which may leads to alteration in fatty acids profile and proinflammatory cytokine expression. Thus, the aim of the present study was to further our understanding about the high incidence of mastitis around parturition by comparison of plasma fatty acid profile and mammary inflammation indicators at different reproductive stages. A total of 47 female rats were included in the present study. After mating, all the pregnant and non-pregnant rats began to receive the same experimental diet. Blood samples were collected at day 1 and 14 of gestation as well as day 3 postpartum.Mammary samples were collected at day 14 of gestation and day 3 postpartum from pregnant and nonpregnant rats. The results showed that rats at d 3 postpartum had greater(P < 0.05) plasma concentrations of non-esterified fatty acids(NEFA), arachidonic acid(ARA) and docosahexaenoic acid(DHA) as well as ARA: eicosapentaenoic acid(EPA) ratio than those at d 14 of gestation. The mRNA abundances of interleukin-1 β(IL-1β), tumor necrosis factor-α(TNF-α),IL-8 and xanthine oxidoreductase(XOR) in mammary of the pregnant rats were greater(P < 0.05) than those in age-matched non-pregnant rats.Rats at d 3 postpartum had higher(P < 0.05) protein expression levels of IL-1β and TNF-α as well as meloperoxidase(MPO) activity and polymorphonuclear neutrophils(PMN) prevalence than those at d 1 of gestation. The rats at d 3 postpartum also had greater(P < 0.05) IL-1β and MPO activity than those at d 14 of gestation. The results indicated that elevated mammary expression of proinflammatory cytokines and XOR as well as altered fatty acid profile around parturition might facilitate the recruitment of neutrophils into mammary glands.

Spliceosomal GTPase Eftud2 regulates microglial activation and polarization

Elongation factor Tu GTP binding domain protein 2(Eftud2)is a spliceosomal GTPase that serves as an innate immune modulator restricting virus infection.Microglia are the resident innate immune cells and the key players of immune response in the central nervous system.However,the role of Eftud2 in microglia has not been reported.In this study,we performed immunofluorescent staining and western blot assay and found that Eftud2 was upregulated in microglia of a 5xFAD transgenic mouse model of Alzheimer’s disease.Next,we generated an inducible microglia-specific Eftud2 conditional knockout mouse line(CX3CR1-CreER;Eftud2^(f/f) cKO)via Cre/loxP recombination and found that Eftud2 deficiency resulted in abnormal proliferation and promoted anti-inflammatory phenotype activation of microglia.Furthermore,we knocked down Eftud2 in BV2 microglia with siRNA specifically targeting Eftud2 and found that Eftud2-mediated regulation of microglial proinflammatory/anti-inflammatory phenotype activation in response to inflammation might be dependent on the NF-κB signaling pathway.Our findings suggest that Eftud2 plays a key role in regulating microglial polarization and homeostasis possibly through the NF-κB signaling pathway.

Cryptotanshinone ameliorates cladribine-induced cognitive impairment in rats

Objective:To evaluate the neuroprotective effect of cryptotanshinone against cladribine-induced cognitive impairment in rats.Methods:Rats were administered with cladribine(1 mg/kg,p.o.)and cryptotanshinone(10 and 20 mg/kg,i.p.)for four weeks.Behavioral tests such as Morris water maze and elevated plus maze were conducted to check memory impairment caused by cladribine.On day 29,all rats were sacrificed,and the brains were separated for estimation of neuroinflammatory factors,biochemical parameters,neurotransmitters,Aβ(1-42),blood-brain barrier permeability,nuclear factor erythroid 2-related factor 2(Nrf2),and brain-derived neurotrophic factor(BDNF).Results:Treatment with cryptotanshinone dose-dependently enhanced spatial memory,improved the levels of neurotransmitter and antioxidant enzymes,and suppressed proinflammatory cytokine release.Cryptotanshinone also decreased Aβ(1-42)accumulation and increased the levels of Nrf2 and BDNF in the hippocampus.Additionally,the histopathological results showed that cryptotanshinone reduced cladribine-induced neuronal death in the hippocampus.Conclusions:Cryptotanshinone exhibits a promising neuroprotective effect against cladribine-induced cognitive impairment in preclinical studies,and may be a potential phytochemical for the treatment and management of cognitive impairment.

Apigenin ameliorates diabetic neuropathy in rats by modulating the TLR4/MyD88 signaling pathway

Objective:To determine the neuroprotective effects of apigenin against streptozotocin(STZ)-induced diabetic neuropathy(DN).Methods:To induce DN,Wistar rats(150-200 g)were administered with STZ(55 mg/kg,i.p.).Then they were randomly assigned to various groups,viz.,normal,diabetic control,insulin(10 IU/kg,s.c.),apigenin(5,10,and 20 mg/kg,p.o.),and insulin(10 IU/kg)plus apigenin(20 mg/kg,p.o.).Various behavioral,biochemical,and molecular markers[tumor necrosis factor-alpha(TNF-α),interleukin(IL)-1β,IL-6,Toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),and nuclear factor erythroid 2-related factor 2(Nrf2)]were assessed.Results:Apigenin(10 and 20 mg/kg,p.o.)substantially reduced plasma glucose levels,lipid profile,aspartate transaminase,alanine transaminase,glycated hemoglobin,and neural advanced glycation end products in STZ-induced DN rats(P<0.05).After apigenin intervention,STZ-induced changes in food and water intake,body weight,urine output,allodynia,hyperalgesia,and insulin levels were markedly improved(P<0.05).Neural antioxidant enzymes(superoxide dismutase and glutathione)and Na+K+ATPase activity were also considerably elevated(P<0.05)while the level of lipid peroxidation was diminished following apigenin therapy(P<0.05).Furthermore,apigenin markedly upregulated the Nrf2 mRNA level while downregulating the mRNA expressions of TNF-αand ILs and the protein expressions of TLR4 and MyD88(P<0.05).STZ-induced histological abnormalities in the sciatic nerve were also improved by apigenin treatment.Conclusions:Apigenin exerts its neuroprotective effect by modulating the inflammatory and oxidative stress pathways via regulating the TLR4-MyD88 signaling pathway.

Kynurenine pathway of tryptophan metabolism in pathophysiology and therapy of major depressive disorder

Serotonin deficiency in major depressive disorder(MDD)has formed the basis of antidepressant drug development and was originally attributed to induction of the major tryptophan(Trp)-degrading enzyme,liver Trp 2,3-dioxygenase(TDO),by cortisol,leading to decreased Trp availability to the brain for serotonin synthesis.Subsequently,the serotonin deficiency was proposed to involve induction of the extrahepatic Trp-degrading enzyme indoleamine 2,3-dioxygenase(IDO)by proinflammatory cytokines,with inflammation being the underlying cause.Recent evidence,however,challenges this latter concept,as not all MDD patients are immune-activated and,when present,inflammation is mild and/or transient.A wide range of antidepressant drugs inhibit the activity of liver TDO and bind specifically to the enzyme,but not to IDO.IDO induction is not a major event in MDD,but,when it occurs,its metabolic consequences may be masked and overridden by upregulation of kynurenine monooxygenase(KMO),the gateway to production of modulators of immune and neuronal functions.KMO appears to be activated in MDD by certain proinflammatory cytokines and antidepressants with anti-inflammatory properties may block this activation.We demonstrate the ability of the antidepressant ketamine to dock(bind)to KMO.The pathophysiology of MDD may be underpinned by both the serotonin deficiency and glutamatergic activation mediated respectively by TDO induction and N-methyl-D-aspartate receptor activation.Inhibition of TDO and KMO should be the focus of MDD pharmacotherapy.

Clostridium butyricum alleviates intestinal low-grade inflammation in TNBS-induced irritable bowel syndrome in mice by regulating functional status of lamina propria dendritic cells

BACKGROUND Irritable bowel syndrome (IBS) is one of the most common functional gastroenterological diseases characterized by abnormal visceral sensitivity and lowgrade inflammation. The role of Clostridium butyricum (C. butyricum) in reducing intestinal low-grade inflammation via immune pathways has been well defined. However, the detailed mechanisms of the effects of C. butyricum on intestinal mucosal immunity, especially on immune cells of the lamina propria, remain unclear. Dendritic cells (DCs), which are important immune cells, secrete proinflammatory cytokines (IL-1β, IL-6, and others) and express T cell immunoglobulin and mucin domain-3 (TIM3), promoting proliferation and activation of DCs, and mediating Th1 and Th17 inflammatory responses. AIM To investigate the role of DCs in the development of IBS in a rat model and to understand the regulation of DCs after C. butyricum intervention. METHODS An IBS animal model was established using C57BL/6 mice, and C. butyricum was continuously administered via the intragastric route to simulate different intestinal immune states. Intestinal visceral hypersensitivity and histopathology were assessed using the abdominal withdrawal reflex (AWR) test and hematoxylin & eosin (H&E) staining, respectively. The expression of proinflammatory cytokines (IL-1β and IL-6) and TIM3 was analyzed by Western blot analysis and real-time PCR. Flow cytometry was applied to analyze the quantity, function, and membrane molecule TIM3 of the lamina propria dendritic cells (LPDCs). The regulatory effect of C. butyricum was verified in bone marrowderived dendritic cells by in vitro experiments. RESULTS The secretion of proinflammatory cytokines (IL-1β and IL-6) in mice with IBS was significantly increased compared with that of the control group, which suggested that the intestinal mucosa in mice with IBS was in a low-grade inflammatory state. The expression of CD11C+CD80+ and CD11c+TIM3+ in intestinal LPDCs in mice with IBS increased significantly. Meanwhile, the cytokines (IL-1β and IL-6) were significantly reduced after the intervention with probiotic C. butyricum. The amount and function of LPDCs and the TIM3 on the surface of the LPDCs were decreased with the alleviation of the intestinal inflammatory response. CONCLUSION The results suggest that C. butyricum regulates the amount and functional status of LPDCs in the intestinal mucosa of mice with IBS, and therefore modulates the local immune response in the intestine.

Preinduced intestinal HSP70 improves visceral hypersensitivity and abnormal intestinal motility in PI-IBS mouse model

Objective:To investigate the impact of the preinduced intestinal heat shock protein 70(HSP70)on the visceral hypersensitivity and abnormal intestinal motility in a post-infectious irritable bowel syndrome(PI-IBS) mouse model.Methods:Eighty-four female C57BL/6 mice were randomly assigned to four groups:control group(n=21) and induction+PI-IBS group(n=21),PI-IBS group(n=21) and induction group(n=21).The mice in PI-IBS group were infected in vivo with trichinella spiralis by oral administration.The visceral hypersensitivity and intestinal motility were evaluated respectively with abdominal withdrawal reflex and colon transportation test.The intestinal HSP70 protein and mRNA level was measured by Western blot and realtime PCR.Meanwhile,the intestinal proinflammatory cytokines IL-10 and TNF-α level was detected by ELISA.Results:Compared with their counterparts in PI-IBS group,the animals in the Induction+PI-IBS group show significantly increased intestinal level of HSP70 and obviously ameliorative clinical tigurcs.including abdominal withdrawal reflex score,intestine transportation time and Bristol scores(P<0.05).Meanwhile,the intestinal post-inflammatory cytokines remarkably changed,including increased IL-10 level and decreased TNF-αlevel(P<0.05).Conclusions:Intestinal IISP70 may play a potential protective role through improving the imbalance between the intestinal post-inflammatory and anti-inflammatory cytokines in PI-IBS.

Nrf2 and Snail-1 in the prevention of experimental liver fibrosis by caffeine

AIM:To determine the molecular mechanisms involved in experimental hepatic fibrosis prevention by caffeine(CFA).METHODS:Liver fibrosis was induced in Wistar rats by intraperitoneal thioacetamide or bile duct ligation and they were concomitantly treated with CFA(15 mg/kg per day).Fibrosis and inflammatory cell infiltrate were evaluated and classified by Knodell index.Inflammatory infiltrate was quantified by immunohistochemistry(anti-CD11b).Gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction for collagenⅠ?(Col-1),connective tissue growth factor(CTGF),transforming growth factorβ1(TGF-β1),tumor necrosis factor alpha(TNF-α),interleukin-1(IL-1),IL-6,superoxide dismutase(SOD)and catalase(CAT).Activation of Nrf2 and Snail-1 was analyzed by Westernblot.TNF-αexpression was proved by enzyme-linked immunosorbant assay,CAT activity was performed by zymography.RESULTS:CFA treatment diminished fibrosis index in treated animals.The Knodell index showed both lower fibrosis and necroinflammation.Expression of profibrogenic genes CTGF,Col-1 and TGF-β1 and proinflammatory genes TNF-α,IL-6 and IL-1 was substantially diminished with CFA treatment with less CD11b positive areas.Significantly lower values of transcriptional factor Snail-1 were detected in CFA treated rats compared with cirrhotic rats without treatment;in contrast Nrf2was increased in the presence of CFA.Expression of SOD and CAT was greater in animals treated with CFA showing a strong correlation between mRNA expression and enzyme activity.CONCLUSION:Our results suggest that CFA inhibits the transcriptional factor Snail-1,down-regulating profibrogenic genes,and activates Nrf2 inducing antioxidant enzymes system,preventing inflammation and fibrosis.