TNF-alpha Expression Research Articles

Structure, antioxidant and immunomodulatory activity of a polysaccharide extracted from Sacha inchi seeds

In this study, a novel water-soluble polysaccharide (PVLP-1) was extracted and purified from Sacha inchi (Plukenetia volubilis L.) seeds and the structure, antioxidant and immunomodulatory activity of PVLP-1 were investigated. PVLP-1 (144 kDa) consisted of glucose (69.76%), mannose (14.86%), arabinose (10.53%), galactose (2.42%), ribose (1.23%), rhamnose (0.27%) and xylose (0.93%). PVLP-1 displayed characteristic polysaccharide bands in Fourier transform NMR spectra and infrared. The primary structure of PVLP-1 was a heteropolysaccharide with a backbone of (1 → 6)-linked glucose, sidechains of (1 → 4)-linked mannose, (1 → 4)-linked glucose and (1 → 3, 6)-linked mannose and a residue unit of →1)-linked arabinose as revealed the methylation analysis. PVLP-1 possessed good water-holding capacity (WHC), oil-holding capacity (OHC) and antioxidant capacities. Besides, PVLP-1 induced the proliferation of RAW264.7 cell and enhanced the expression of inflammatory cytokines IL-6, TNF-alpha(TNF-α) and IL-1 beta (IL-1β). The present study indicated that PVLP-1 possessed immune-enhancing bioactivities and could be functional food or adjuvant drug to improve biological immunity of immunodeficiency diseases and hypoimmunity.

P0123THE IMPORTANCE OF PPAR-ALPHA IN AKI TO CKD TRANSITION IN NEPHROPATHY INDUCED BY FOLIC ACID

Abstract Background and Aims PPAR-alpha is a nuclear receptor which plays major role in the regulation of lipid metabolism, activation of PPAR-alpha has been shown to have beneficial effects in renal diseases. Acute kidney injury (AKI), even if followed by renal recovery, is a risk factor for the future development of chronic kidney disease (CKD) and end-stage renal disease (ESRD). Our main objective was to determine the importance of PPAR-alpha in AKI to CKD transition in folic acid induced nephropathy. Method Male C57/Bl6 and PPARKO (C57Bl6 background) mice were divided in 4 groups, Control, Folic Acid, Gemfibrozil + Folic Acid and PPARKO + Folic Acid. Animals has been treated with single dose of Folic Acid (250mg/kg i.p) and Gemfibrozil (150mg/kg gavage) euthanized after 48 hours for AKI assessment and 28 days for CKD. Renal parameters, histology, real time PCR were performed to investigate renal injury, inflammation and fibrosis. Results After 48 hours of folic acid inducing AKI, PPARKO mice showed decreased urea levels (Folic Acid: 178,9±25,2 PPARKO: 92,4±15,5), less tubular injury (Folic Acid: 0,61±0,05 PPARKO: 0,21±0,03), lower mRNA expression of NGAL (Folic Acid; 32,2±7,67 PPARKO 3,74±1,71), KIM-1 (Folic Acid: 671,8±170,2 PPARKO: 43,4±29,7) and TNF-alpha (Folic Acid: 2,81±0,58 PPARKO: 0,67±0,14), while PPAR-alpha activation with gemfibrozil showed to have no effects in AKI. After 28 days of folic acid inducing CKD. PPARKO showed same levels of injury than folic acid alone treated mice, however PPAR-alpha activation with gemfibrozil decreased urea levels (Folic Acid: 79,2±3,2 Gemfibrozil: 46,1±3,8), showed less tubulointerstitial fibrosis (Folic Acid: 0,60±0,05 Gemfibrozil: 0,18±0,02) and lower urine protein/creatinine ratio (Folic Acid: 5,48±0,58 Gemfibrozil: 2,53±0,12), Conclusion PPAR-alpha deletion protects against AKI induced by folic acid but did not showed protection in chronic phase, in the other hand ppar-alpha activation with gemfibrozil did not protect in AKI but show to be effective in CKD.

The methylation status of TNF-α and SOCS3 promoters and the regulation of these gene expressions in patients with Behçet’s disease

Introduction: The aim of this study was to evaluate the methylation status of TNF-α and SOCS3 promoters in patients with BD and compare them with a healthy group.Method: This was a case–control study, in which 47 subjects with BD and 61 individuals as the control participated. Blood samples were collected from all the participants. Then, PBMCs were isolated using the Ficoll method and methylation of considered sites was investigated using the qMS-PCR technique after DNA extraction by the rapid genomic DNA extraction method and its analysis with Nano-drop.Results: The methylation and expression of TNF-α showed that the methylation level significantly declined in the patient in comparison with the healthy (p < 0.05). Moreover, the results on the mean expression showed that it significantly increased in the patient group, as compared with the healthy group (p < 0.05). In addition, the expression of the SOCS3 gene was not significantly different between the patients and healthy subjects while the level of SOCS3 methylation was significantly higher in the patient group than that in the healthy group (p < 0.05).Discussion: The present study revealed that the gene expression of TNF-alpha increased in BD patients, suggesting that TNF-alpha likely has a role in the pathogenesis of BD.

Antidepressant Effect of Cinnamon (Cinnamomum burmannii) Bark Extract in Chronic Stress-Induced Rats

BACKGROUND: Depression is a psychiatric disorder that has become a serious health problem in the past decade. This disorder is characterized by prolonged dysphoric mood, and in more severe condition would result in decreased self-care and even life-threatening action. Serotonin is believed to play a role in the regulation of mood elation in depressive disorders. Decreased levels of serotonin in the hippocampus will cause an increased dopamine in mesolimbic dopamine neuronal cells. An effective and commonly used drug is the selective serotonin reuptake inhibitor, namely, fluoxetine. However, this agent has so many side effects, one of them is erectile dysfunction. In order to find the better treatment, exploration and discovery of therapeutic modalities need to pursued using natural materials.&#x0D; AIM: This study aimed to explore and evaluate antidepressant effects of cinnamon (Cinnamomum burmannii) extract (CE).&#x0D; METHODS: A total of 30 male Wistar rats were obtained from Eureka Research Laboratory (Palembang, Indonesia). Cinnamon simplisia was obtained from the Institute for Research and Testing of Traditional Medicine, Tawangmangu, Central Java, Indonesia. Rats were induced using chronic mild stress (CMS). CMS was a form of stress induction performed on experimental animals continuously, for 4 weeks. Forced swimming test (FST) was a test conducted to assess mobility in animal model. After induction for 4 weeks, rats were randomly divided into six groups which each contained five rats: Normal control group, CMS group (negative control), CMS + fluoxetin (Fluox 1: mg/kg), CMS + CE 25 mg/kg, the CMS + CE 50 mg/kg, and the CMS + CE 100 mg/kg. Treatment with fluoxetine or CE was given for 14 days intragastrically using gastric sonde. After treatment and FST, organ evacuation was performed and followed by immunohistochemistry and enzyme-linked immunosorbent examination.&#x0D; RESULTS: This study showed that CE with dose of 25 mg/kg BW to dose 100 mg/k BW could reduce the duration of immobility when compared to the CMS group. Clinically, CE possessed the potential to reduce the duration of immobility and potentially reduce symptoms of depression. Histologically, CE showed the potential to improve serotonin levels in the hippocampus with increasing doses. Tumor necrosis factor (TNF)-alpha expression in the hippocampus as a marker of inflammation had increased in the CMS group. CE was able to reduce the expression of TNF-alpha compared to the CMS group.&#x0D; CONCLUSION: CE possessed antidepressant efficacy by inhibiting the inflammatory process in the hippocampus so it was able to optimally increase serotonin levels in the hippocampus.

Vitamin D (VD3) antioxidative and anti-inflammatory activities: Peripheral and central effects

Vitamin D (VD3, cholecalciferol), besides its role on bone calcium homeostasis, has also been shown to present anti-inflammatory actions. The objectives of the present work were to further extend these findings, focusing onVD3action mechanisms at the molecular level and onits central and peripheral effects. For that, VD3 antinociceptive and mainly anti-inflammatory activities were evaluated by acute models of nociception (formalin test) and inflammation (carrageenan-induced paw edema), in mice pretreated orally for 7 days with VD3 (0.5 and 1.0 mg/kg). Afterwards, the edematous paws were evaluated by immunohistochemical assays for TNF-alpha. In addition, brains from mice pretreated with VD3, at the same conditions, were harvested for iNOS andCOX-2 immunohistochemical (IHC) assays. The anti-inflammatory effect of VD3 on human neutrophil degranulation was evaluated by the release of myeloperoxidase (MPO) activity, as well as by the reactive oxygen species production. VD3 significantly reduced the licking time in the formalin test, at the second phase (inflammatory pain). VD3 also reduced the edema volume and the number of polymorphonuclear (PMN) cells, as well as the TNF-alpha expression in the edematous paws, compared with the control group. Furthermore, VD3 significantly decreased iNOS and COX-2 expressions in brain areas, such as hippocampus and prefrontal cortex, and inhibited the degranulation of activated neutrophils by the reduction of ROS production and MPO release. Based in these results, VD3 presents anti-inflammatory and antioxidative effects, manifested at peripheral and central sites as showed in the present work for the first time.

Abstract A29: Differential regulation of apoptosis and senescence by the miR-449 family microRNAs and its implications in tumor microenvironments of NSCLC

Abstract MicroRNAs (miRNAs) play important roles in various biologic processes, and aberrant miRNA expression is one of the mechanisms of tumorigenesis. Previous studies indicate that oncogenic HPV infection interrupts the expression of tumor-suppressive miR-34a and contributes to the development of cervical cancer. In addition, the traditional Chinese medicine Gynostemma pentaphyllum (Thunb.) Makino may exert its pharmacologic effects through modifying the gut microbiota by regulating hepatic miR-34a expression. MiR-449 miRs are members of the tumor suppressor miR-34 family and are evolutionarily conserved. In this study, we demonstrated that, in lung cancer cells, ectopic expression of miR-449 family, including miR-449a, miR-449b, and miR-449c, decreased the levels of several G1 phase proteins including cyclin D1, cyclin D3, CDK4, and CDK6 via directly targeting their mRNA’s 3’-UTR. We also observed a differential induction of apoptosis vs. senescence by the miR-449 cluster. While miR-449a and miR-449c induced senescence, miR-449b induced apoptosis and sensitized cancer cells to apoptotic stimuli, such as serum deprivation. Interestingly, among this cluster miRNAs, only miR-449a could induce significant senescence-associated secretory phenotype (SASP), e.g., expression of IL-8, IL-6, IL-1b, and TNF-alpha. Moreover, we also found that miR-449a increased NF-kB phosphorylation at S276, but not phosphorylation site of S536, and promoted the expression of IL-8 mRNA. In this regard, the gut microbiota has been indicated to induce proinflammatory cytokines such as IL-8 to influence the outcomes of cancer patients. In addition to MRX34 (a miR-34a mimic) used in clinical trials, miR-449 family may also potentially become a new therapeutic candidate for NSCLC and other solid tumors’ treatment. Citation Format: Chun-Yu Cho, Shih-Ying Chung, Shuang-En Chuang. Differential regulation of apoptosis and senescence by the miR-449 family microRNAs and its implications in tumor microenvironments of NSCLC [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr A29.

Effects of TNF‐Alpha on TGF‐Beta Expression in Human Skeletal Muscle Cells: A Cell Model for Inflammation

Approximately 25.3 million adults suffer from chronic pain, often caused by chronic inflammatory conditions. We hypothesize that chronic inflammation negatively alters myofascial tissue structure and function. To better understand this process and test our hypothesis, we are developing a cell model to examine chronic inflammation in myofascial tissue. The goal of this investigation is to determine if chronic inflammation changes cell signaling of the myofascial tissue and lead to fibrosis. Human skeletal muscle (HSkM) cells and human fibroblasts make up the myofascial tissue. Therefore, we will first examine the relationship between the chronic inflammatory mediator, TNF‐alpha, and HSkM cell expression of TGF‐beta. We find that TNF‐alpha increases expression of TGF‐beta in HSkM cells changing the cell phenotype to a fibrotic state.

A polyphenol-rich Calafate (Berberis microphylla) extract rescues glucose tolerance in mice fed with cafeteria diet

Abstract The establishment of a chronic inflammatory state in the adipose tissue contributes to obesity-associated insulin resistance. Hence, disrupting the inflammatory response elicited by obesity remains a relevant target to tackle the modern-world pandemic. We evaluated the anti-inflammatory and insulin-sensitizing effect of Calafate (Berberis microphylla) by producing and characterizing a polyphenol-pure Calafate extract (PPCE). C57BL/6 mice fed with cafeteria diet for 14 weeks were administered PPCE (50 mg/Kg/day) for 4 weeks. PPCE administration rescued glucose tolerance and insulin-elicited AKT phosphorylation in white adipose tissue of diet-induced insulin-resistant mice. Furthermore, the cafeteria diet-induced expression of TNF-alpha and F4/80 was attenuated by PPCE administration, suggesting that PPCE rescues insulin sensibility by ameliorating the obesity-associated inflammatory state. Altogether, our data shows that Calafate represents a natural source of polyphenols with glucose tolerance-improving properties in vivo, suggesting a potential use of PPCE as a complementary tool against insulin resistance.

Anti-inflammatory effect from extracts of Red Chinese cabbage and Aronia in LPS-stimulated RAW 264.7 cells.

A chronic inflammatory environment facilitates tumor growth and proliferation. Fruits and vegetables are important sources of anthocyanins, polyphenols, and other biologically active substances that can favorably affect the pathogenesis of cancer. The objective of the study was to investigate the anti‐inflammatory effects of Red Chinese cabbage (RC) and mixture of commercial Red Chinese cabbage leaves and Aronia fruits (ARC) in LPS‐stimulated RAW 264.7 cells. The RAW 264.7 cells were cultured and measured the cytotoxicity by using an MTT assay. The inflammatory markers, such as nitrite, IL‐6, and TNF‐alpha expression, were evaluated using ELISA, and protein expression of inflammatory markers like iNOS and COX‐2 was analyzed using Western blot. MTT assays showed that pretreatment of RAW 264.7 cells with RC and ARC did not change cell growth or cytotoxicity. We also found that ARC extracts reduced inflammation‐related biomarker (TNF‐a, IL‐6, and NO) production and gene expression (iNOS, COX‐2). Our results suggested that ARC has good anti‐inflammatory properties compared with RC that maybe used as potential nutrients for treating inflammatory diseases.

Functional characterization of two ecto-nucleoside triphosphate diphosphohydrolase 2 genes in Japanese flounder (Paralichthys olivaceus) head kidney macrophages

Ecto-nucleoside triphosphate diphosphohydrolases (ENTPDases) are pivotal regulators of extracellular ATP-mediated purinergic immune signaling. ENTPDase2 is a member of the cell surface-bound ecto-nucleoside triphosphate diphosphohydrolase (ENTPDase) protein family that hydrolyzes extracellular nucleoside 5′-triphosphates and nucleoside 5′-diphosphates. However, the immune relevance of ENTPDase2 in fish has not been elucidated. In the present study, from a comparative immunological perspective, we functionally characterized two ENTPDase2 transcript variants (namely ENTPDase2 and ENTPDase2a) from Japanese flounder (Paralichthys olivaceus). Sequence analysis indicates that the deduced Japanese flounder ENTPDase2 and ENTPDase2a proteins possess two conserved transmembrane domains and five apyrase conserved regions that are present in ENTPDase family proteins. However, these proteins only share 54% amino acid sequence identity. Tissue expression analysis revealed that both ENTPDase2 and ENTPDase2a mRNA transcripts are ubiquitously expressed in all examined Japanese flounder tissues, whereas ENTPDase2 is dominantly expressed in blood and ENTPDase2a is abundantly expressed in muscle. Immune challenge experiments showed that ENTPDase2 and ENTPDase2a were significantly upregulated by both inflammatory stimulation and Edwardsiella tarda infection. In addition, the expression of ENTPDase2 and ENTPDase2a was modulated by extracellular ATP (eATP) stimulation in a dose-dependent manner. Furthermore, immunolocalization and functional studies demonstrated that both ENTPDase2 and ENTPDase2a are functional glycosylated plasma membrane proteins. However, ENTPDase2a exhibits greater activity in the hydrolysis of eATP than ENTPDase2 and ENTPDase1 proteins. Finally, knockdown of the ENTPDase2 gene by small interfering RNA significantly upregulated the expression of eATP-induced proinflammatory cytokines IL-1beta, TNF-alpha and G-CSF in Japanese flounder head kidney macrophages, while knockdown of ENTPDase2a only upregulated eATP-induced IL-1beta expression. Taken together, our findings suggest that the two functional Japanese flounder ENTPDase2 isoforms play an essential role in the downregulation of eATP-induced proinflammatory cytokine expression in fish by degrading the available ATP levels in the extracellular milieu.

Adipose-Derived Molecules-Untouched Horizons in Alzheimer's Disease Biology.

The global incidence of Alzheimer’s disease (AD) is on the rise with the increase in obesity and metabolic disease epidemic. Obesity is co-morbid with the increase in mass of adipose tissue, which secretes numerous molecules that are biologically important. Obesity and its associated conditions are perhaps involved in the causative pathway of AD. Immunologically important cytokines such as IL-1β, IL-10, and IL-18, which are released by adipose tissue, are also found to be associated with AD. Besides, the expression of IL-6, IFNγ, and TNF alpha are also associated with AD. Ang-I and Ang-II are found to mediate the progression of AD. Complement factors B, C4b, and H are differentially expressed in AD. Overall, several adipocyte-derived cytokines are found to be dysregulated in AD, and their role in AD remains to be studied. The induction of autophagy is a very promising strategy in the treatment of AD. A variety of adipose-derived molecules have been shown to modulate autophagy. However, very little literature is available on the role of adipose-derived molecules in inducing autophagy in microglial cells of AD. Understanding the role of adipose-derived molecules in the development of AD, especially in the induction of autophagy, would open up new avenues in devising strategies for the treatment of AD.

Ganoderma Lucidum Polysaccharides Ameliorates Hepatic Steatosis and Oxidative Stress in db/db Mice via Targeting Nuclear Factor E2 (Erythroid-Derived 2)-Related Factor-2/Heme Oxygenase-1 (HO-1) Pathway.

BackgroundType 2 diabetes mellitus (T2DM) and its comorbidities, including obesity, hypertension, and hyperlipidemia, are commonly associated with non-alcoholic fatty liver disease (NAFLD). Ganoderma lucidum polysaccharide (GDLP) is one of the central bioactive components in Ganoderma lucidum with anti-inflammatory, antioxidant, and hepatoprotective properties. However, the effect and mechanisms of GDLP in hepatic steatosis remain largely unknown. In the present study, we aimed to investigate the function of GDLP in hepatic steatosis and the underlying mechanism.Material/MethodsIn this study, male db/db mice were received with a high-fat diet (HFD) to investigate the effect of GDLP in T2DM-induced hepatic steatosis. The biological characteristics of the hepatic steatosis were evaluated through the detection of clinical indicators, including biochemical parameters, histopathology, and related cytokine levels. Additionally, the protein expression levels of Nrf2 (nuclear factor E2 (erythroid-derived 2)-related factor-2) signaling pathway were investigated by using western blotting and immunohistochemical staining.ResultsThe levels of food/water intake, body weight, fasting blood glucose, plasma lipids, urinary biomarkers, hepatic lipid accumulation, and tumor necrosis factor (TNF)-α were observably decreased in GDLP-treated db/db mice. Additionally, administration of GDLP increased the expression of various antioxidases, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), whereas it reduced the level of malonaldehyde (MDA). Furthermore, GDLP was significantly promoted protein expression level of Nrf2 and its downstream target gene HO-1 (heme oxygenase-1) while decreased TNF-α expression.ConclusionsThese results indicate that GDLP against T2DM-induced hepatic steatosis, oxidative stress, and inflammation by improving the Nrf2/HO-1 signaling pathway in db/db mice, suggesting the GDLP may serve as an effective strategy for in fatty liver treatment.

Abstract TP272: Selective Inhibition of Histone Deacetylase 3 Decreases Cerebral Edema and Blood-Brain Barrier Leakage

Introduction: Histone deacetylase 3 (HDAC3) has been implicated as neurotoxic in several neurodegenerative conditions. However, its role in ischemic stroke has not been thoroughly explored. In this study, we examined HDAC3 expression after stroke and tested whether selective inhibition of HDAC3 decreases cerebral edema and blood-brain barrier (BBB) leakage after ischemia. Methods: To examine HDAC3 expression over time, adult male Wistar rats (n=6/group) were subjected to middle cerebral artery occlusion (MCAO) and sacrificed at different time points after stroke. Another set of adult male Wistar rats was subjected to 2-h MCAO, and randomly selected animals were treated i.p. with either vehicle (1% Tween 80) or a selective HDAC3 inhibitor (RGFP966, 10 mg/kg) at 2 and 24 h after MCAO. Behavioral tests were performed, and infarct volumes and cerebral edema were measured. Evans blue dye (EBD) assay was employed to assess BBB leakage. Immunostaining for HDAC3, CD31, claudin-5, occludin, and ZO-1; and Western Blot for TNF-alpha and matrix metalloproteinase-9 (MMP9) were performed. Results: Total and neuronal HDAC3 significantly increased in the peri-infarct cortex as early as 3 hours after ischemia. HDAC3 levels continued to be significantly elevated at 3 days and returned to baseline at 7 days. Selective HDAC3 inhibition improved functional outcome (p=0.01) and reduced infarct volumes (p&lt;0.001). Compared to vehicle control, RGFP966 treatment significantly decreased cerebral edema (p&lt;0.001) as well as BBB leakage, as measured by EBD assay. Immunostaining showed that RGFP966 significantly increased the expression of tight junction proteins (claudin-5, occludin, and ZO-1) in the peri-infarct area compared to vehicle control group. Western Blot showed that RGFP966 treatment significantly decreased the expression of TNF-alpha and MMP9 in the ischemic brain. Conclusions: HDAC3 is upregulated in the ischemic brain as early as 3 hours after stroke. Early administration of a selective HDAC3 inhibitor improves neurological functional outcome and decreases infarct volume, cerebral edema, and BBB leakage. BBB protection by selective inhibition of HDAC3 is mediated in part by upregulation of tight junction proteins and attenuation of TNF-alpha and MMP9.

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages.

Calreticulin (CALR) has anti-tumor effects by increasing dendritic cell maturation and tumor antigen presentation. However, whether CALR affects macrophages and modulates progression of acute respiratory distress syndrome/acute lung injury (ARDS/ALI) remains unknown. In this study, we discovered that CALR protein was highly expressed in the mice with LPS-induced ALI and CALR expression level was positively correlated to the severity of ALI. Commercial anti-CALR antibody (aCALR) can neutralize recombinant CALR (rCALR) and suppress the expression of TNF-alpha and IL-6 in the rCALR-treated macrophages. Blocking CALR activity by intraperitoneal (i.p.) administration of aCALR significantly suppressed ALI, accompanied with lower total cell counts, neutrophil and T cell infiltration in bronchoalveolar lavage (BAL) and lung tissues. The expression of CXCL15, IL-6, IL-1beta, TNF-alpha, and CALR were significantly reduced, in association with more polarization of Siglec F+CD206+M2 subtype macrophages in the aCALR-treated mice. Pre-depletion of circulating monocytes did not abolish the aCALR-mediated suppression of ALI. Further analysis in bone marrow-derived macrophages (BMDMs) showed that aCALR suppressed the expression of CD80, IL-6, IL-1beta, IL-18, NLRP3, and p-p38 MAPK; but enhanced the expression of CD206 and IL-10. In addition, we observed more expression and phosphorylation of STAT6 in the aCALR-treated BMDM. Lack of STAT6 resulted in comparable and slightly higher expression of CALR, TNF-alpha and IL-6 in the aCALR-treated STAT6-/- BMDMs than the untreated cells. Therefore, we conclude that CALR is a novel biomarker in the evaluation of ALI. Blocking CALR activity by aCALR effectively suppressed ALI independent of circulating monocytes. Siglec F+CD206+M2 subtype macrophages and p38 MAPK/STAT6 signaling pathway played important role in the immune regulation of aCALR. Blocking CALR activity is a promising therapeutic approach in the treatment of ARDS/ALI.

Level of Expression of BDNF and TNF-Alpha in the Somatosensory Cortex and its Effect on Social Interaction in Autism Model Rats Induced by Valproic Acid

Level of Expression of BDNF and TNF-Alpha in the Somatosensory Cortex and its Effect on Social Interaction in Autism Model Rats Induced by Valproic Acid

Characteristics of neutrophil extracellular traps in patients with periodontitis and gingivitis.

We sought to compare the characteristics and clinical significance of neutrophil extracellular traps in gingival samples from patients with periodontitis and those with gingivitis. The clinical indexes of gingival samples from patients with periodontitis and gingivitis were measured; the expression of TNF-alpha and IL-8 was measured by real-time fluorescent quantitative PCR; and the expression of TLR-8 and MMP-9 was measured by western blotting assays. Chemotaxis, phagocytosis and phagocytic activity of neutrophils were measured. Compared with the healthy group, the expression of TNF-α and IL-8 in the periodontitis group and the gingivitis group increased significantly (p < 0.05), and TNF-α in the gingivitis group was significantly lower than that in the healthy group (p < 0.05). The expression of IL-8 in the periodontitis group was significantly higher than that in the periodontitis group (p < 0.05). Furthermore, the expression of TLR-8 and MMP-9 in the periodontitis group was different from that in the gingivitis group and the healthy group, and the expression of TLR-8 and MMP-9 in the gingivitis group was significantly different from that in the healthy group (p < 0.05). In addition, the neutrophil mobility index in healthy people was 3.02 ± 0.53, that in the periodontitis group was 2.21 ± 0.13, and that in the gingivitis group was 2.31 ± 0.12. In conclusion, the chemotaxis of neutrophils in gingival samples of patients with periodontitis and gingivitis was decreased, the phagocytotic ability and activity of neutrophils were reduced, and the release of the extracellular trap-releasing inducible factors TNF-alpha and IL-8 also declined.

Effect of cysteine peptidase inhibitor of Eudiplozoon nipponicum (Monogenea) on cytokine expression of macrophages in vitro

The gills of the common carp, whose mucosal surface belongs to the key defence mechanisms of piscine immunity, can be infested with both the larval and adult stage of Eudiplozoon nipponicum (Monogenea). Although on their own, monogeneans do not considerably compromise their hosts’ health status, fish with epithelial barriers damaged in parasite feeding and attachment sites are at an increased risk of bacterial challenge with possible harmful consequences. Several studies suggest that helminth parasites of teleost fish evade and manipulate host immune system via their excretory-secretory products, but our knowledge of these processes in the monogeneans is limited. Cysteine peptidase inhibitors (CPI), which are found in the secretions of numerous parasites, often induce immunosuppression by subverting Th1 mechanisms and drawing the immune system towards a Th2/Treg response. We employed the qPCR to test the effect of recently characterised CPI of E. nipponicum (rEnStef) on the mRNA expression of pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 produced by porcine macrophages in vitro. After an initial preincubation with rEnStef, we stimulated the macrophages using LPS. By inducing a Th1 pro-inflammatory response, we imitated the immune reaction during a bacterial challenge in tissue damaged by the feeding and attachment of E. nipponicum. We observed a significant dose-dependent downregulation of the expression of TNF-α and IL-10 cytokines. The observed suppression of TNF-alpha expression by rEnStef could result in decreased pathogen control, which might in turn lead to increased rates of secondary bacterial infections in fish infected by E. nipponicum.

NLRC4 inflammasome has a protective role on inflammatory bone resorption in a murine model of periodontal disease.

There is virtually no information on the role of NLRC4 inflammasome on bone resorption and inflammation associated with periodontitis. Bacterial-associated experimental periodontitis was induced in wild-type (WT) and Nlrc4-KO C57BL/6 mice. 3 μL of a 1 × 109 UFC/mL PBS suspension of heat-killed Gram-negative bacteria were injected (3x/week for 4 weeks) directly into the gingival tissues of WT and Nlrc4-KO mice (n = 6/genotype). Control animals were injected bilaterally (3x/week for 4 weeks) in the same sites with the same volume of the PBS vehicle. Alveolar bone resorption was quantified by μCT. Inflammatory infiltrate in the gingival tissues was assessed qualitatively in H&E-stained slides and by the detection of a pan-leukocyte marker (CD45) and a neutrophil marker (Ly6G) using immunofluorescence. Modulation of Rankl, Mmp-13, Tnf-a, Il-6 and Il-10 expression in the gingival tissues was determined by RT-qPCR. Osteoclastogenesis was assessed in vivo by biochemical staining for TRAP. The relevance of NLRC4 for RANKL-induced osteoclastic differentiation and activity was investigated in vitro using bone marrow-derived macrophages from WT and Nlrc4-KO mice. Bone resorption was significantly greater in Nlrc4-KO mice; however there were no differences between WT and Nlrc4-KO mice on osteoclast numbers and on the inflammatory infiltrate. In vitro, osteoclast activity was significantly enhanced in Nlrc4-deficient macrophages; whereas RANKL-induced differentiation was not affected. Expression of the selected candidate genes was also similarly increased by the induction of experimental periodontal disease, except for the expression of Tnf-alpha and Il-10, which was already significantly higher in the gingival tissues of Nlrc4-KO mice. We conclude that NLRC4 inflammasome has a protective role on inflammatory bone resorption in this experimental model. Furthermore, the bone-sparing effect may be related with the modulation of osteoclast activity.

Can dietary jujube (Ziziphus jujuba Mill.) fruit extract alter cutaneous mucosal immunity, immune related genes expression in skin and growth performance of common carp (Cyprinus carpio)?

The present study investigates the effect of dietary Ziziphus jujube fruit extract [ZJFE] on skin mucus non-specific immune parameters as well as mRNA levels of immune related gene in the skin of common carp fingerlings. An eight-week feeding trial was performed using different levels of dietary ZJFE (0, 0.25, 0.5 and 1%). At the end of the trial skin mucus immune parameters (total Ig, lysozyme and protease activity), cytokines genes (il1b, il8, il10 and tnf-alpha) expression in skin and growth performance were studied. The result showed highest and lowest skin mucus total Ig were observed in fish fed diet containing 0.5% ZJFE and control group, respectively (P < 0.05). There were no significant difference among treatments regarding skin mucus lysozyme activity (P >0.05). It should be noted that, feeding on 0.5 and 1% ZJFE significantly increased skin mucus protease activity (P <0.05). Likewise, gene expression studies in skin showed significant increase of il1b expression in fish fed 0.5% ZJFE compared other treatments (P <0.05). Also, il8 gene was noticeably up-regulated in 0.5 and 1% treatments compared to the control group (P <0.05). While there were no significant difference between 0.25% JFE treatment and control in case of relative il10 gene expression (P >0.05), feeding on diets containing 0.5% or 1% ZJFE significantly down-regulated il10 gene (P <0.05). Our study indicated that relative expression of tnf-alpha gene significantly increased in treated groups (P <0.05). Also, feeding on ZJFE supplemented diet improved growth performance parameters. Overall, this experiment demonstrated the potentially useful effects of ZJFE on skin mucosal immunity and performance of common carp fingerlings.

Royal Jelly (Bee Product) Decreases Inflammatory Response in Wistar Rats Induced with Ultraviolet Radiation.

BACKGROUND:Ultraviolet (UV) radiation damages human skin by triggering various types of cellular damage, several main factors involved are nuclear-related factor 2 (Nrf2), nuclear factor kappa-light-chain-enhancer of activated B cells (nF-kB) and pro-inflammatory cytokine, TNF alpha. Royal jelly (RJ) possesses the effect of protecting DNA and tissue against oxidative damage.AIM:This study aimed to assess the efficacy of RJ as a protector of ultraviolet radiation, by assessing endogenous anti-oxidant expression (Nrf2), transcription factors (Nf-kB) and proinflammatory cytokines (TNF alpha).METHODS:This study was an experimental study with post-test control group design. Thirty Wistar rats were induced by exposing 40 Watt UV-B lamps for 2 hours/day in 14 days. The rats were grouped into groups with RJ cream application with doses of 2.5%, 5%, and 10%, negative control with vaseline, and normal control. Examination of Nrf2 and NF-kB levels was carried out by ELISA. Quantitative analysis to obtain the percentage of TNF alpha expression on the tissue was entered into the ImageJ® program. Bivariate analysis was carried out by the T-test.RESULTS:Nrf2 levels elevated following the increase of RJ dose, with the highest level was at RJ 10%. Nf-kB levels decreased following the increase of RJ dose, with the lowest level was at RJ 10%. TNF alpha expression was reduced in groups of RJ in various doses. Increased dose resulted in a more diminished level of TNF alpha.CONCLUSION:Royal jelly cream application protected the skin from UV radiation by increasing cellular antioxidants and suppressing inflammatory cascade.