Levels Of Anti-inflammatory Cytokines Research Articles

S580 Effect of Probiotics on Serum Cytokines in Irritable Bowel Syndrome With Gulf War Illness

Introduction: Gulf war illness (GWI) is a multisystem disorder of unknown etiology. There is high prevalence of IBS among Veterans with GWI. There is inconsistent evidence that low grade chronic gastrointestinal (GI) inflammation may play a part in the pathogenesis of IBS and GWI. We examined the blood cytokines profile of Veterans with IBS and GW illness before and after treatment with probiotics. Methods: In a randomized double-blind placebo controlled trial we enrolled Veterans with IBS (Rome III criteria) and two or more symptoms of GWI. Veterans were randomized to receive probiotic (De Simone Formulation; formally known as VSL#3) or identical placebo. Blood was collected at enrollment and after 8-week treatment with probiotics. A multiplexed immunoassay was used to assess the serum concentration of 13 cytokines/inflammatory markers: interferon (IFN)-γ; interleukins (ILs)-1β, 2, 4, 5, 6, 8, 10, 12, 13, and 17; tumor necrosis factor (TNF-α, IL-2 receptor. The cytokine levels were compared to reference intervals established on 120 healthy controls. Results: Data was analyzed from 42 Veterans who provided baseline and 11 Veterans who provided both pre and post treatment blood samples for cytokine analysis. The serum pro- and anti-inflammatory cytokines levels were no different between IBS with GWI and healthy controls. There was no change in pro- or anti-inflammatory cytokines after treatment with probiotics (Figure). Conclusion: The level of pro- and anti-inflammatory cytokines were no different in Veterans with IBS and GWI compared with healthy controls and the levels do not alter after treatment with probiotics. The role of cytokines in pathophysiology, and as a biomarker of treatment of IBS and GWI is likely complex and remains unknown.Figure 1.: Comparison of Pre and Post treatment with probiotics cytokines (mean±SE)

Immunomodulation of MiRNA-223-based nanoplatform for targeted therapy in retinopathy of prematurity.

Retinopathy of prematurity (ROP) is characterized by pathological angiogenesis and associated inflammation in the retina and is the leading cause of childhood blindness. MiRNA-223 (miR-223) drives microglial polarization toward the anti-inflammatory phenotype and offers a therapeutic approach to suppress inflammation and consequently pathological neovascularization. However, miRNA-based therapy is hindered by the low stability and non-specific cell-targeting ability of delivery systems. In the present study, we developed folic acid-chitosan (FA-CS)-modified mesoporous silica nanoparticles (PMSN) loaded with miR-223 to regulate retinal microglial polarization. The FA-CS/PMSN/miR-223 nanoparticles exhibited high stability and loading efficiency, achieved targeted delivery, and successfully escaped from lysosomes. In cultured microglial cells, treatment with FA-CS/PMSN/miR-223 nanoparticles upregulated the anti-inflammatory gene YM1/2 and IL-4RA, and downregulated the proinflammatory genes iNOS, IL-1β, and IL-6. Notably, in a mouse oxygen-induced retinopathy model of ROP, intravitreally injected FA-CS/PMSN/miR-223 nanoparticles (1μg) decreased the retinal neovascular area by 52.6%. This protective effect was associated with the reduced and increased levels of pro-inflammatory (M1) and anti-inflammatory (M2) cytokines, respectively. Collectively, these findings demonstrate that FA-CS/PMSN/miR-223 nanoparticles provide an effective therapeutic strategy for the treatment of ROP by modulating the miR-223-mediated microglial polarization to the M2 phenotype.

ALTERED PLASMA CYTOKINE CONCENTRATIONS ARE ASSOCIATED WITH THE CLINICAL SEVERITY OF PATIENTS WITH COVID-19

IntroductionThe novel Coronavirus (SARS-CoV-2), responsible for severe acute respiratory syndrome, has emerged as a threat to humans since December 2019, and the search for a better understanding of the pathophysiology of coronavirus disease 2019 (COVID-19) and its definitive treatment is still in progress.ObjectiveTo evaluate the plasma pro- and anti-inflammatory cytokines in COVID-19 patients and their associations with the disease severity and outcome.MethodsReverse-Transcriptase Polymerase Chain Reaction (RT-PCR)-confirmed COVID-19 unvaccinated patients at the Hospital de Clínicas (HC), UNICAMP, Campinas, SP, were enrolled. Clinical and laboratory data were extracted from the medical records, and the plasma cytokines levels were quantified using LUMINEX and ELISA.ResultsThere were 154 COVID-19 patients (99 survivors and 55 non-survivors) with male:female of 1.4:1, and a median age of 60 years. The non-survivors were older than survivors (65 vs. 55 years, p < 0.0001); and coronary artery disease and autoimmunity, disease severity, and oxygen therapy, intensive care, and intubation were associated with mortality. Non-survivors had higher leukocyte and neutrophil counts, and RDW and lower lymphocyte count at diagnosis. Non-survivors had higher levels of pro-inflammatory (TNF-α, IL-6, IFN-γ, CCL3, IL-17/IL-17A, IL-8, G-CSF, CCL2/MCP-1) and anti-inflammatory (IL-1ra and IL-27) cytokines, but lower TGF-β levels than the survivors. TNF-α levels were positively correlated with all studied cytokines except TGF-β, while TGF-β levels were negatively correlated with TNF-α, IL-6, CCL3, G-CSF, and IL-27. IL-27 levels were significantly correlated with all the cytokines except IL-37 and IL-17E. More than half (55.2%) of our patients had severe COVID-19, 18.8% had moderate, 16.2% had critical, 5.2% had mild, and 4.5% were asymptomatic. Majority of the patients (68.2%) required ICU care and had higher TNF-α, IL-6, IL-8, IL-17, CCL3, CCL2, IL-1ra, and IL-27 than others. 59.7% of the patients required endotracheal intubation and had higher TNF-α, IL-6, IL-8, CCL3, CCL2, and IL-1ra than those who did not have intubation. TNF-α, IL-6, and IL-8 had the highest Area Under the Receiver Operating Characteristics (AUROC) curve, sensitivity, and specificity for predicting mortality in these COVID-19 patients.Discussion and conclusionThe altered levels of pro- and anti-inflammatory cytokines support the role of SARS-CoV-2 in inducing cytokine storm, and higher concentrations seen in the deceased patients meant a more severe storm. Also, the increased leukocytes and neutrophils in our patients could have led to the release of reactive oxygen species, and end-organ damage, thus leading to poor outcomes. This study showed that the levels of these cytokines could be used as markers of mortality in COVID-19. It is possible to suggest that TNF, IL-6, and IL-8 levels at diagnosis could be efficient predictors of fatal outcomes in COVID-19 patients. If properly measured at diagnosis, these markers could be useful for triaging and predicting the outcome of COVID-19, thus guiding the treatment of the COVID-19.FundingCNPq (#190374/2017-9), CAPES, FAPESP and FAEPEX (#338619).

In situ hydrogel capturing nitric oxide microbubbles accelerates the healing of diabetic foot.

Diabetic foot ulcer (DFU) is a devastating complication in diabetes patients, imposing a high risk of amputation and economic burden on patients. Sustained inflammation and angiogenesis hindrance are thought to be two key drivers of the pathogenesis of such ulcers. Nitric oxide (NO) has been proven to accelerate the healing of acute or chronic wounds by modulating inflammation and angiogenesis. However, the use of gas-based therapeutics is difficult for skin wounds. Herein, therapeutic NO gas was first prepared as stable microbubbles, followed by incorporation into a cold Poloxamer-407 (P407) solution. Exposed to the DFU wound, the cold P407 solution would rapidly be transformed into a semisolid hydrogel under body temperature and accordingly capture NO microbubbles. The NO microbubble-captured hydrogel (PNO) was expected to accelerate wound healing in diabetic feet. The NO microbubbles had an average diameter of 0.8±0.4μm, and most of which were captured by the in situ P407 hydrogel. Moreover, the NO microbubbles were evenly distributed inside the hydrogel and kept for a longer time. In addition, the gelling temperature of 30% (w/v) P407 polymer (21°C) was adjusted to 31°C for the PNO gel, which was near the temperature of the skin surface. Rheologic studies showed that the PNO gel had mechanical strength comparable with that of the P407 hydrogel. The cold PNO solution was conveniently sprayed or smeared on the wound of DFU and rapidly gelled. In vivo studies showed that PNO remarkably accelerated wound healing in rats with DFU. Moreover, the sustained inflammation at the DFU wound was largely reversed by PNO, as reflected by the decreased levels of proinflammatory cytokines (IL-1β, IL-6 and TNF-α) and the increased levels of anti-inflammatory cytokines (IL-10, IL-22 and IL-13). Meanwhile, angiogenesis was significantly promoted by PNO, resulting in rich blood perfusion at the DFU wounds. The therapeutic mechanism of PNO was highly associated with polarizing macrophages and maintaining the homeostasis of the extracellular matrix. Collectively, PNO gel may be a promising vehicle of therapeutic NO gas for DFU treatment.

Pro- and anti-inflammatory cytokine profiles in Plasmodium Falciparum-infected individuals from Baiyeku, Ikorodu, Lagos, Nigeria

Available evidence indicates that the various stages of the malaria parasite life cycle elicit specific immune responses of which the relative levels of pro-inflammatory cytokines are key to disease progression, killing the parasite and mediating disease outcomes. This study investigated T-cell response in malaria. Four hundred and sixty-two participants were screened in a community survey of Plasmodium falciparum malaria in Baiyeku, Lagos, Nigeria. P. falciparum parasitaemia was determined by microscopy while the serum levels of IL-10, IFNγ and TNFα were determined by Enzyme-linked immunosorbent assay (ELISA). A total of 70 (15.2 %) participants were microscopically positive for P. falciparum of which 70% were females, 30% were males while children aged 1-17 years were 65.7%. The geometric mean parasite density (GMPD) was significantly (p=0.001) higher among females than males. The GMPD of participants &lt;5 years of age was also significantly (p=0.001) higher than other age groups. About 46.8% of the participants were underweight (Body Mass Index, BMI &lt; 18.5) and also had the highest parasite intensity. The TNFα, IFNγ and IL-10 levels were significantly (p&lt; 0.05) higher in the infected than the uninfected participants. IFN-γ values were significantly (p=0.014) elevated among the symptomatic than the asymptomatic participants while there was no significant difference (P&gt;0.053) in the levels of TNF-α and IL-10 (P&gt;0.093) between the symptomatic and asymptomatic participants. The prevalence of P. falciparum obtained in this study area which is endemic to malaria is 15.2% suggesting a significant reduction of the disease over time due to awareness of the disease in the community. The levels of pro-inflammatory cytokines (IFN-γ and TNF-α) in this study were lower due to the down-regulatory action of the anti-inflammatory cytokines (IL-10). These findings suggest that higher levels of anti-inflammatory cytokines, especially IL-10 levels may contribute to the pathogenesis of uncomplicated malaria.

The impact of aquatic exercise programs on the systemic hematological and inflammatory markers of community dwelling elderly: A randomized controlled trial.

Evidence shows that physical exercise is important in maintaining an efficient immune system during ageing. However, there are few studies that test the impact of aquatic exercise programs on the immune system. This study aims to analyze the impact of different physical exercise programs in aquatic environment on the systemic hematological and inflammatory markers of community dwelling elderly. One hundred and two elderly were randomly allocated into four groups: a continuous aerobic exercise group (AerG) (n = 25, 71.44 ± 4.84 years); an interval aerobic exercise group (IntG) (n = 28, 72.64 ± 5.22 years); a combined exercise group (ComG) (n = 29, 71.90 ± 5.67 years); a control group (CG) (n = 20, 73.60 ± 5.25 years). The AerG, IntG and ComG participants took part in three different aquatic exercise programs over a 28-weeks period. The CG participants maintained their usual routines during the same time period. Blood samples were collected from all participants in order to access hematologic indicators, by means of cell count, and the inflammatory profile by ELISA. After 28 weeks, significant differences were found for several hematologic variables in the AerG, IntG and ComG with increases in mean corpuscular hemoglobulin (MCH), mean corpuscular hemoglobulin concentration (MCHC), and hemoglobulin (Hb). Decreases in TNF-α levels were found for all exercising groups. An increase in IL-10 levels, granulocytes to lymphocytes ratio (GLR) and a decrease in the TNF-α/IL-10 ratio, were found for the IntG. For the ComG decreases were also found for the TNF-α, IL-1ß/IL-1ra ratios. The present study suggests that aquatic exercise programs were able to improve the inflammatory profile of the participants. Those in the exercise intervention groups showed a shift towards lower pro-inflammatory levels while the non-exercising group showed the opposite behaviour. The IntG and the ComG aquatic exercise programs appeared to be more effective than the AerG program in decreasing chronic low-grade inflammation by mediating the production of higher levels of anti-inflammatory cytokines. However, the differences found between the exercising groups were small and may not have clinical significance.

Pattern recognition receptor CD14 gene polymorphisms in alcohol use disorder patients and its Influence on liver disease susceptibility.

BackgroundAlcohol use disorders (AUDs) leading to liver disease is major concern over other spectrum of disorder. Excessive alcohol consumption resulting in leaky gut syndrome is attributed to alcohol-induced liver injury through portal translocation of bacterial endotoxin. Susceptibility to alcoholic liver disease (ALD) in AUD patients could be dependent upon genes responsible for inflammation and alcohol metabolism. The pattern recognition receptor CD14 gene is a major player in endotoxin-mediated inflammation and susceptibility to ALD. This study investigated the genetic association of CD14 polymorphisms and other mechanisms relevant to altered inflammatory responses leading to ALD.MethodsPatients with alcohol use disorder with ALD (n = 128) and without liver disease (ALC, n = 184) and controls without alcohol use disorder (NALC, n = 152) from North India were enrolled. The CD4 gene polymorphisms in the North Indian population were evaluated by RFLP and sequencing. Secretory CD14 (sCD14), LBP, TLR4, MD2, TNFα, IL1b, IFNγ, IL6, IL10, and IL4 levels in serum were measured by ELISA among groups. The influence of polymorphisms on CD14 gene promoter activity and circulatory bacterial DNA level was determined.ResultsThe CD14 gene promoter and exonic region SNPs were found to be monomorphic, except for SNP rs2569190 for the North Indian population. The genetic association of SNP rs2569190(C/T) with the risk of developing ALD was found significant for TT genotype [ORTT, 95% CI = 2.19, 1.16–4.13 for ALD vs. ALC and OR, 2.09, 1.18–3.72 for ALD vs. NALC]. An increased sCD14 level was observed in AUD patients compared to NALC control. Increased levels of LBP, TLR4, TNFα, IL1β, IFNγ, and IL6 and reduced levels of MD2, IL10, and IL4 were observed among the ALD patients compared to the other two control groups. Elevated levels of pro-inflammatory and reduced levels of anti-inflammatory cytokines were observed in the risk genotype TT groups of ALD patients and the ALC group compared to NALC. Promoter activity was observed in the intronic region flanking SNPs and risk genotype can influence reporter activity, indicating CD14 gene expression.ConclusionEnhanced CD14 expression associated with inflammatory responses increases susceptibility to ALD in the TT genotype of AUD patients.

Investigation of the Gastroprotective Effect of Betaine-Homocysteine Homeostasis on Oxidative Stress, Inflammation and Apoptosis in Ethanol-Induced Ulcer Model

Background: There is a growing interest in the use of natural compounds for the treatment of gastric ulcers. The multifunctional roles of betaine in various diseases make this natural substance a favorable pre-drug for ulcer treatment. This study aims to determine the competence of betaine in gastroprotection against ethanol-induced damage and to explore underlying mechanisms considering its effects on liver and kidney activity and blood parameters. Methods: Wistar albino rats were orally treated with vehicle (distilled water) or betaine (250 mg/kg) for twenty-one days and then ulcer formation was induced by ingestion of 75% ethanol. Gastric mucosal damage was evaluated by gross examination and histopathological analysis. Homocysteine levels, lipid peroxidation, total antioxidant status (TAS), total oxidant status (TAS), antioxidant enzymes and pro-inflammatory and anti-inflammatory cytokines levels were assessed by enzyme-linked immunosorbent assay (ELISA) or immunohistochemistry. Furthermore, routine biochemical tests were performed and hematological parameters were analyzed. Results: Betaine ameliorated any gastric mucosal damage and reduced homocysteine levels significantly. The TOS and malondialdehyde (MDA) levels were decreased while the TAS, glutathione (GSH) levels and catalase (CAT) activity were increased upon the betaine treatment. Betaine reduced apoptosis by regulating Bax and Bcl-2 levels, however, it did not alter inflammatory mediators. Additionally, betaine improved serum potassium (K+) and blood urea nitrogen (BUN) levels, whereas it increased alanine aminotransferase (ALT) levels and impaired hematological parameters. Conclusions: Altogether, these data illustrated that betaine exhibits a gastroprotective effect against ulcers through the homocysteine pathway by modulating oxidative stress in the gastric tissue; however, its systemic effects should not be ignored.

Chuanzhitongluo regulates microglia polarization and inflammatory response in acute ischemic stroke

Background and purposeChuanzhitongluo (CZTL), a traditional Chinese medicine mixture, is used in the recovery period of acute ischemic stroke (AIS), and effectively improves the prognosis of AIS patients. This study aims to evaluate whether CZTL regulates microglia polarization and inflammatory response to reduce brain damage in the acute phase of AIS. MethodsA mouse model of AIS was prepared by the photochemical method. Cerebral infarct volume was detected by 2,3,5-Triphenyltetrazolium chloride (TTC) staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to assess neuronal apoptosis. Gene expression profile change was explored by Gene chip. Inflammatory factors were analyzed by Protein microarray. The Immunofluorescence double-labeling assay was executed to elucidate the effects of CD16+ / Iba-1+ and CD206+ / Iba-1+ in the peripheral area of cerebral ischemia. ResultsResults revealed that CZTL treatment alleviated the neurological impairment, reduced cerebral infarct volume, and inhibited neuronal apoptosis. CZTL altered gene expression profiles, which indicate that CZTL may be involved in regulating neuroinflammation. CZTL restrained inflammatory responses by down-regulated pro-inflammatory cytokines expression and enhanced anti-inflammatory cytokines level. Further experiments demonstrated that CZTL inhibited the activation of NLRP3 inflammasome, which decreasing the inflammatory response. In addition, CZTL promoted the transformation of microglia from M1 to M2 phenotype. ConclusionsThese results indicate that CZTL alleviates neuroinflammation and brain damage after AIS in mice, which may be mediated by modulating microglia polarization.

ДИНАМИКА ПОКАЗАТЕЛЕЙ ЦИТОКИНОВОГО ПРОФИЛЯ ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ПАРОДОНТИТЕ

Abstract. The molecular level of physiological processes in the context of studying the pathogenesis of diseases is the basis for their diagnosis and treatment. Thus, the features of the pathogenesis of periodontitis are the direct participation of pro-inflammatory cytokines in the development of alterative inflammation. The study of the cytokine profile is relevant for both diagnostic and therapeutic purposes, since the dynamics of indicators of inflammatory mediators reflects the degree of activity and specificity of pathological reactions present in periodontitis. The purpose of the study Study of cytokine profile indicators in experimental periodontitis. Materials and methods. In an experiment on 50 white non-linear rats weighing 190-280 g, the model of experimental periodontitis proposed by Shkolnaya K.D., Atrushkevich V.G. was reproduced. (Patent RU No. 2625295 dated 07/12/2017). Assessment of the dynamics of the cytokine status of blood serum was carried out by the level of pro-inflammatory (IL-1β, TNF, IL-6, IL-17), anti-inflammatory (IL-4, IL-10) and regulatory (IL-2) cytokines using enzyme-linked immunosorbent assay using reagent kit "Bender MedSystems". Results. The study revealed the presence of an imbalance in the cytokine system, characterized by a violation of the physiological balance between pro-inflammatory and anti-inflammatory cytokines with a predominance of the former. Hyperproduction of cytokines persisted throughout the experiment with an increase in the above uncoupling by day 25, which indicates the formation of an immunoinflammatory process in the periodontium. Conclusion. The data obtained during the study confirm the presence of an imbalance of the cytokine network, which indicates the formation and progression of the inflammatory process of the periodontal complex, and also necessitates the introduction of drugs that inhibit the synthesis of pro-inflammatory mediators into complex therapy.

Is hepatitis-B immunization effective during chronic liver fibrosis? Investigation of secretory and cellular immune responses on an experimental model

Objective Adults with end-stage of chronic liver diseases have lower antibody titers after hepatitis-B vaccination. We have less amount of knowledge about the effect of non-viral cause chronic liver fibrosis on vaccination. In this study, we investigated the effect of non-viral chronic liver fibrosis on hepatitis B vaccine and the effect of tetanous toxoid co-administration at the level of humoral and cellular immune responses in an experimental model. Methods Hepatitis B vaccine was administered either alone or in combination with tetanus toxoid in thioacetamide-induced fibrotic BALB/c mice. Fibrosis level was determined by Knodell scoring. Anti-HBsAg, biochemical parameters, inflammatory (IL-1β, TNF-α), and anti-inflammatory (IL-10) cytokine levels were investigated in serum samples by automated systems and ELISA; respectively. Frequencies of activated lymphocytes were determined in flow cytometer. Results Antibody titers significantly decreased after immunization of fibrotic mice. However, co-administration of toxoid significantly elevated antibody titer. The percentage of CD19+CD69+ B lymphocytes was found to be lower in vaccinated fibrotic group compared to vaccinated naive group. Simultaneous administration of toxoid significantly increased the frequencies of CD4+ and CD8+ T cells expressing CD69 and CD127. Interestingly, CD19+CD5+CD1high Breg cells were significantly reduced in the group vaccinated with hepatitis B vaccine and toxoid, simultaneously. The reduction in Breg percentage did not expose a significant decrease in the level of IL-10. Conclusion Non-viral chronic liver fibrosis causes a reduction on specific antibody level after vaccination. Reduction on Breg cell frequency may have an effect on elevation of antibody level after co-administration of tetanus toxoid.

Jatrorrhizine Alleviates DSS-Induced Ulcerative Colitis by Regulating the Intestinal Barrier Function and Inhibiting TLR4/MyD88/NF-κB Signaling Pathway.

Background Ulcerative colitis (UC), a kind of autoimmune disease with unknown etiology, has been troubling human physical and mental health. Jatrorrhizine (Jat) is a natural isoquinoline alkaloid isolated from Coptis Chinensis, which has been proved to have antibacterial, anti-inflammatory, and antitumor effects. Purpose The purpose is to explore the therapeutic effect of Jat on DSS-induced UC and the mechanism of action. Study Design. The UC mice model was induced by 3% DSS in drinking water. The mice were orally administered with Jat (40, 80, 160 mg/kg) for 10 days. Methods The changes in body weight, colon length, spleen wet weight index, disease activity index (DAI), colonic histopathology, and inflammatory factors of serum and colon tissue were analyzed to evaluate the severity of colitis mice. The colon mucus secretion capacity was analyzed by Alcian blue periodic acid Schiff (AB-PAS) staining. Furthermore, protein expressions such as TLR4, MyD88, p–NF–κB-p65, NF-κB-p65, COX-2, ZO-1, and Occludin were detected to elucidate the molecular mechanism of Jat on DSS-induced colitis model. Results The results showed that Jat could significantly alleviate the symptoms, colon shortening, spleen index, and histological damage and restore the body weight in DSS-induced colitis mice. Jat also suppressed the levels of inflammatory cytokines and upregulated the levels of anti-inflammatory cytokines. In addition, Jat repaired the intestinal barrier function by upregulating the level of colonic tight junction (TJ) proteins and enhancing the secretion of mucin produced by goblet cells. Furthermore, Jat could significantly suppress the expression of TLR4, MyD88, p–NF–κB-p65/NF-κB-p65, and COX-2 in colon tissue. Conclusion The results suggested that Jat plays a protective role in DSS-induced colitis by regulating the intestinal barrier function and inhibiting the TLR4/MyD88/NF-κB signaling pathway. This study, for the first time, demonstrates the therapeutic and protective effects of Jat on UC.

Potential Protective Role of Galectin-3 in Patients with Gonarthrosis and Diabetes Mellitus: A Cross-Sectional Study.

Background: Gonarthrosis and diabetes mellitus are two diseases that are increasingly being linked. The aim of this study was to quantify serum levels of Gal-3, pro- and anti-inflammatory cytokines (including their ratios and correlations), and participant’s condition (pain, stiffness, functional limitations) in gonarthrosis patients with and without diabetes mellitus. Methods: A between-subject, cross-sectional experimental design was adopted. Serum levels of TNF-α, IL-6, IL-12, IL-23, IFN-γ, IL-17, IL-10, Gal-3, and WOMAC score were measured. Results: Gonarthrosis patients with diabetes mellitus had significantly (p < 0.05) lower levels of TNF-α, IL-6, IL-12, IL-17, IFN-γ, and Gal-3 compared to gonarthrosis patients without diabetes mellitus. On the other hand, IL-10/TNF-α, IL-10/IL-6, IL-10/IL-12, Gal-3/TNF-α, Gal-3/IL-6, and Gal-3/IL-12 (p = 0.001) were significantly higher (p < 0.05) in gonarthrosis with diabetes mellitus. Moderate–large correlation (p < 0.05) was detected between the serum values of Gal-3 and pro- and anti-inflammatory cytokines, including IL-12 (r = 0.575), IL-10 (r = 0.535), TNF-α (r = 0.306), and IL-23 (r = 0.323). WOMAC index was significantly lower (p < 0.05) in gonarthrosis patients without diabetes mellitus compared to gonarthrosis patients with diabetes mellitus. Conclusions: Correlation between Gal-3 and proinflammatory cytokines and its dominance over proinflammatory cytokines implicate the potential role of Gal-3 in preventing cartilage destruction.

Phytochemical analysis, cytostatic, cytotoxic, and anti-inflammatory effects of Arum palaestinum, Ocimum basilicum, and Trigonella foenum-graecum in human monocytic cell line (THP-1)-derived macrophages

The significance of the crosstalk between pro-inflammatory mediators and carcinogenesis is widely discussed. These mediators play decisive roles at different stages of tumor development, including initiation, promotion, and metastasis. Arum palaestinum Boiss., Ocimum basilicum L., and Trigonella foenum-graecum L. and their crude extracts are traditionally used in the Arab and Islamic herbal medicine to treat a variety of cancers and inflammatory illnesses. Human monocytic cell line (THP-1)-derived macrophages were used to evaluate anti-inflammatory, cytotoxic, and cytostatic effects of the ethanolic plant extracts. Cytotoxic and cytostatic effects were measured with the MTT assay. In addition, the production levels of pro-inflammatory mediators (TNF-α, IL-6, and nitric oxide) and anti-inflammatory cytokine (IL-10) were measured in lipopolysaccharide (LPS)-activated THP-1-derived macrophages in the absence and presence of increasing concentrations of the three plant extracts. The three plant extracts suppressed the production of NO and TNF-α and IL-6, and enhanced the production of IL-10 in LPS-activated THP-1-derived macrophages. In addition, these extracts inhibited the growth of THP-1-derived macrophages in a concentration-dependent manner at nontoxic concentrations. T. foenum-graecum exhibited the highest cytostatic effects with an IC50 of 512 µg/ mL compared to O. basilicum (no cytostatic effects) and A. palaestinum (IC50=1274 µg/mL). Even though more studies are needed to elucidate the mechanisms of observed cytostatic and anti-inflammatory effects, to some extent, these effects could be attributed to the flavonoids, phenolic compounds, and tannin content detected in the plants’ extracts.

Naringenin Attenuates Cognitive Impairment in a Rat Model of Vascular Dementia by Inhibiting Hippocampal Oxidative Stress and Inflammatory Response and Promoting N-Methyl-D-Aspartate Receptor Signaling Pathway.

Vascular dementia (VaD) is the second most common form of dementia globally, yet there are no efficient treatments. Naringenin, a natural flavonoid, exerts antioxidative, anti-inflammatory, and neuroprotective properties; however, its potential effect on VaD remain unclear. Herein, the purpose of present study was to elucidate whether naringenin attenuates cognitive dysfunction in VaD via inhibiting hippocampal oxidative stress and inflammatory response, and promoting N-methyl-D-aspartate receptors (NMDARs) signaling pathway. A rat model of VaD was established by permanent bilateral common carotid artery occlusion [2-vessel occlusion (2VO)]. Behavioral performance analyses results revealed that administration of naringenin improves cognitive impairment in rats with VaD according to the new object recognition test and the Morris water maze test. In addition, naringenin attenuated hippocampal oxidative stress by reducing reactive oxygen species generation, decreasing malondialdehyde content and recombinant reactive oxygen species modulator 1 (Romo-1) expression, and increasing superoxide dismutase and glutathione peroxidase activities in the hippocampus of VaD rats. Moreover, naringenin decreased the proinflammatory cytokines (IL-1β, IL-6, and TNF-α) levels and increased the anti-inflammatory cytokines (IL-10 and IL-4) levels in the hippocampus of 2VO surgery-treated rats, attenuating hippocampal inflammatory response during VaD. Furthermore, naringenin promoted synaptophysin (SYP), postsynaptic density protein 95 (PSD95), N-methyl-Daspartic acid receptor 1 (NR1) and N-methyl-D-aspartate receptor subunit 2B (NR2B) expressions levels in hippocampus of VaD rats. Collectively, these findings indicated thatnaringenin mitigates cognitive impairmentin VaD rats partly via inhibiting hippocampal oxidative stress and inflammatory response and restoring NMDARs signaling pathway.

Xanthones from Securidaca inappendiculata Hassk. attenuate collagen-induced arthritis in rats by inhibiting the nicotinamide phosphoribosyltransferase/glycolysis pathway and macrophage polarization

Securidaca inappendiculata (SI) Hassk. is a traditional medicine used to treat rheumatoid arthritis. Recent studies have reported that macrophages are the primary regulators of joint homeostasis and their polarization is closely related to their metabolic mode. Here, we aimed to investigate the relationship between the joint protective effect of SI’s xanthone-rich fraction (XRF) on collagen-induced arthritis (CIA) in rats and the nicotinamide phosphoribosyltransferase (NAMPT)-glycolysis-polarization axis of macrophages. CIA model rats were treated with oral XRF and therapeutic efficacy was assessed based on arthritis score, degree of paw swelling, histological examination, and immunohistochemical analysis. Serum levels of cytokines, cellular metabolite concentrations, and protein and mRNA expression were determined by enzyme-linked immunosorbent assay (ELISA), western blotting (WB), and quantitative real-time PCR (RT-qPCR), respectively. The effects of dihydroxy-3,4-dimethoxyxanthone (XAN), a representative SI-derived compound, on RAW264.7 macrophages was analyzed in vitro using confocal laser scanning and flow cytometry. We found that XRF treatment significantly alleviated disease severity in CIA model rats. Levels of pro-inflammatory cytokines in the serum and M1 markers in synovium were reduced after XRF treatment, accompanied by an increase in the levels of anti-inflammatory cytokines and M2 markers. Further, XRF significantly suppressed synovial glycolysis by regulating NAMPT. In vitro studies further showed that XAN induced repolarization of lipopolysaccharide (LPS)-induced RAW264.7 macrophages with M1-M2 phenotype. Moreover, XAN negatively regulated glycolysis in the LPS-induced RAW264.7 macrophages in correlation with changes in NAMPT expression. Overall, the findings of this study suggest that the joint protective effects of XRF are achieved by inhibiting the NAMPT/glycolysis pathway and thereby regulating macrophage polarization.

Tolerance Assessment of Atractylodes macrocephala Polysaccharide in the Diet of Largemouth Bass (Micropterus salmoides).

Atractylodes macrocephala polysaccharide (AMP) can enhance antioxidant defense and anti-inflammation, as the tolerance levels of AMP in aquaculture is important for additive utilization. However, the tolerance dose of AMP is unknown. We assess the tolerance levels of AMP in juvenile largemouth bass (3.38 ± 0.11 g) by feeding them a 0, 400, 4000, or 8000 mg/kg AMP supplemented diet for 10 weeks. The 400 mg/kg AMP dose increased growth performance. The Nrf2/Keap1 signaling pathway was activated, as indicated by Keap1 and Nrf2 protein levels in the liver. Enhanced activity of antioxidant enzymes (SOD, GPx), together with increased mRNA levels of antioxidant genes (sod, gpx) and decreased accumulation of reactive oxygen species (ROS) and MDA, was found in the liver, implying the antioxidant effect of AMP. Nutrient absorption was enhanced by AMP, as reflected by the increased length of intestinal villi and microvilli. However, 4000 and 8000 mg/kg AMP induced oxidant stress, as indicated by increased plasma ALT and AST content and decreased mRNA levels of antioxidant genes (sod, gpx) in the liver and intestinal tissues. Inflammatory reactions were also induced by high doses of AMP, as reflected by enhanced levels of pro-inflammatory cytokines (tnfα, nfκb) in the liver, intestinal, and kidney tissues and inhibited levels of anti-inflammatory cytokines (tgfβ, iκb). Histological analysis reveals inflammatory cell infiltration and tissue damage. Thus, the safe tolerance margin of AMP supplement for largemouth bass was 400–4000 mg/kg.

Regulation of microglial activation in stroke in aged mice: a translational study.

Numerous neurochemical changes occur with aging and stroke mainly affects the elderly. Our previous study has found interferon regulatory factor 5 (IRF5) and 4 (IRF4) regulate neuroinflammation in young stroke mice. However, whether the IRF5-IRF4 regulatory axis has the same effect in aged brains is not known. In this study, aged (18-20-month-old), microglial IRF5 or IRF4 conditional knockout (CKO) mice were subjected to a 60-min middle cerebral artery occlusion (MCAO). Stroke outcomes were quantified at 3d after MCAO. Flow cytometry and ELISA were performed to evaluate microglial activation and immune responses. We found aged microglia express higher levels of IRF5 and lower levels of IRF4 than young microglia after stroke. IRF5 CKO aged mice had improved stroke outcomes; whereas worse outcomes were seen in IRF4 CKO vs. their flox controls. IRF5 CKO aged microglia had significantly lower levels of IL-1β and CD68 than controls; whereas significantly higher levels of IL-1β and TNF-α were seen in IRF4 CKO vs. control microglia. Plasma levels of TNF-α and MIP-1α were decreased in IRF5 CKO vs. flox aged mice, and IL-1β/IL-6 levels were increased in IRF4 CKO vs. controls. The anti-inflammatory cytokines (IL-4/IL-10) levels were higher in IRF5 CKO, and lower in IRF4 CKO aged mice vs. their flox controls. IRF5 and IRF4 signaling drives microglial pro- and anti-inflammatory response respectively; microglial IRF5 is detrimental and IRF4 beneficial for aged mice in stroke. IRF5-IRF4 axis is a promising target for developing new, effective therapeutic strategies for the cerebral ischemia.

Modulatory Effects of Leave and Fruit Extracts of Ficus sycomorus on Cytostatic and Inflammatory Mediators in Monocultures and Co-cultures of Human Keratinocyte (HaCat) and Human Monocyte (THP-1) Cell Lines

Background: Ficus sycomorus is one of the recommended antipsoriatic medicinal plants in the traditional Greco-Arab herbal medicine. However, the knowledge on its action mechanisms is limited.&#x0D; Aims: Cytotoxic, cytostatic, and anti-inflammatory effects of water/ethanolic extracts of Ficus sycomorus leaves and fruits were evaluated to test their role in the traditionally known antipsoriatic properties.&#x0D; Place and Duration of Study: All the experiments were done in the Department of Biology and Biotechnology, Arab American University-Palestine in 2020. Plants were collected from the Northern region of the West Bank/Palestine during the fall months in 2019 and given (Voucher code: Pharm-PCT-1030) at An-Najah National University.&#x0D; Methodology: MTT assay was used to evaluate cytostatic and cytotoxic effects of Ficus sycomorus extracts in human skin keratinocyte cell line (HaCaT), human monocytic cell line (THP-1)-derived macrophages, and their co-cultures. Commercial ELISA kits were applied to measure the cytokine levels.&#x0D; Results: Both extracts exhibited cytostatic effects with IC 50 of 656 μg/mL and 886 μg/mL for HaCat and co-culture, respectively. Leaves and fruits extracts significantly reduced dose-dependently the LPS-induced NO production by THP-1-derived macrophages from 65 μM to 19 μM and 16 μM, respectively. The fruit extracts showed higher effects than the leaf extracts and reduced the TNF-α levels from 709 pg/mL to 208 pg/mL. The fruit extracts increased the production levels of IL-10 from 74 pg/mL to 90 pg/mL.&#x0D; Conclusion: Ficus sycomorus extracts probably exert their antipsoriatic effects through cytostatic effects and modulation of the production levels of pro-inflammatory and anti-inflammatory cytokines.

Abstract P3073: Altered Mitochondrial Mg 2+ Homeostasis In Liver Links Lipid Metabolism, Inflammation And Atherogenesis

Although evidences show the role of Mg 2+ in lipid metabolism and atherosclerosis, the molecular mechanism by which loss of Mg 2+ homeostasis promotes cardiovascular disease (CVD) is not known. Mg 2+ homeostasis is regulated by mitochondria and Mrs2 is the only known molecular machinery associated with mitochondrial Mg 2+ influx. To begin elucidating the importance of Mg 2+ homeostasis in lipid metabolism and CVD we made liver-specific Mrs2 KO mouse (Mrs2 Δhep ). Liver being the central hub of lipid metabolism, we asked whether loss of Mrs2 alters lipid metabolism and lipoprotein profile. Our results show increased accumulation of hepatic triacylglycerol (TG) and cholesterol in Mrs2 Δhep . Consistent with increased TG/cholesterol, plasma cholesterol and oxidized low density lipoprotein (ox-LDL) levels significantly differed between control and KO. Because ox-LDL contribute to atherosclerosis, we hypothesized altered hepatic lipid metabolism to induce inflammation, immune imbalance and accelerate lesions through modified atherogenic lipoprotein and cytokine profile. Quantification of plasma cytokines and chemokines were performed using the proteome profiler. Because T cells are critical drivers and modifiers of atherosclerosis, we quantified the lymphocyte and monocyte subsets by flow cytometry. Because T regulatory cells have anti-antherogenic roles, we studied the effect of ox-LDL on Tregs. Our results show increased and decreased levels of inflammatory and anti-inflammatory cytokines respectively in Mrs2 Δhep . In Mrs2 Δhep stimulation with ox-LDL showed an increased antigen specific proliferation of T-cells. We also observed increased and decreased population of Th17 and Tregs respectively. The decrease in Tregs population was attributed to increased apoptosis. In conclusion our observation suggests that high levels of ox-LDL in Mrs2 Δhep to induce inflammation and inflammatory cell expansion accompanied by apoptosis in Tregs. Our results conclude dysregulated Mg 2+ homeostasis to alter lipid metabolism and atherogenesis by inducing inflammation and compromised Tregs population. Our study will be the first of its kind to establish a triangular molecular link between Mg 2+ homeostasis, lipid metabolism, and atherogenesis.