Reduced Expression Of TNF-α Research Articles

Potential of Binahong (<i>Anredera cordifolia</i> [Tenore] Steen) in Reducing TNF-<i>α</i> Expression on Regeneration of Pancreas <i>β</i> Cells on White Rats (<i>Rattus norvegicus</i>) Diabetes Mellitus Models

Diabetes mellitus (DM) is a metabolic disorder characterized by increased blood glucose levels (hyperglycemia) due to impaired insulin secretion and/or increased insulin resistance. Providing binahong leaf steeping (Anredera cordifolia [Tenore] Steen) is one alternative treatment for people with diabetes mellitus using medicinal plants. This research was conducted to find out the right concentration by administering binahong leaf steeping to reduce hyperglycemia in STZ induced rat and to analyze administration of binahong leaf steeping in reducing TNF-α expression in pancreatic β cell regeneration. This study used 25 white rats (Rattus novergicus) which were divided into 5 groups: group 1, negative control (K-), group 2, positive control (K+), groups 3, 4, and 5. With STZ induction, each is given a dose of binahong leaf steeping, 50.4 mg/180ml of water; 100.8 mg/180ml of water and 151.2 mg/180ml of water for 14 days. The results showed that the administration of binahong leaf steeping to STZ induced rat could reduce glucose rats in a row by 130.00, respectively; 117,23 and 112,45 and steeping binahong leaves (Anredera cordifolia [Ten] S) significantly influence the reduction in TNF-α expression in pancreatic β cell regeneration. Thus, this study can be concluded that the right dose to reduce hyperglycemic and regeneration of pancreatic β cells produced by STZ induced rats is 151.2 mg/180ml of water and statistically influences in reducing TNF-α expression in pancreatic β cell regeneration.

Effects of Curcumin on Biological Behavior and NF-κB/TNF-α Pathway in Mice with Metastatic Bone Pain of Breast Cancer Induced by Walker 256 Cells

Background. The active ingredient curcumin of traditional Chinese medicine was selected as the research object to investigate the possible mechanism of breast cancer metastatic bone pain in mouse walker 256 cells and the effect of curcumin on the NF-κB/TNF-α pathway in order to provide a new idea for clinical treatment of breast cancer metastatic bone pain. Methods. By establishing an animal model of breast cancer bone metastasis in walker 256 cells, the biological behavior of nude mice was observed on the 8th day after successful modeling. Meanwhile, the low dose group, middle dose group and high dose group of mice were given 15 mg/kg, 25 mg/kg, 50 mg/kg of curcumin solution intraperitoneally in 21 days, and the right cavity bone and spinal cord distended in mice (L4-L6) tissues were used to detect related factors, Immunohistochemical method was used to detect c-fos in spinal cord. Expression levels of RANK, NF-κB and TNF-α were detected by RT-PCR and Western blot. Meanwhile, serum levels of Cox2, il-6, leukotriene and PGE2 were detected. Results. Observing the biological behavior index of nude mice, we found that the mechanical pain and thermal pain threshold decreased (p Conclusions. On the 8th day after the success of the animal model of breast cancer bone metastasis in Walker 256 cells, abnormal biological behaviors such as heat pain, cold pain sensation and spontaneous hyperalgesia were observed. Further studies have found that the increased expression of rank on osteoclasts induced up-regulated expression of NF-κB and c-fos, induced expression of TNF-α gene, and could induce synthesis and release of leukotriene, PGE2 through direct activation of cyclooxygenase, inflammatory media IL-6 cascade reaction, resulting in pathological pain and hypersensitivity. Traditional Chinese medicine active ingredient curcumin could reduce RANK expression of osteoclast, inhibit cell NF-κB and spinal cord c-fos activity, reduce TNF-α expression, inhibit Cox2 activity, and reduce the synthesis and release of inflammatory factors leukotriene and PGE2, thus exerting its analgesic effect, which provides new ideas and methods for clinical treatment of metastatic bone pain in breast cancer.

The effect of purple sweet potato (Ipomea batatas L.) extract gel toward TNF-α expression and skin collagen density on rats (Rattus norvegicus) with open wound models

Open wound is a damage to the skin or tissues under the skin that can be caused by sharp objects or fights between animals. Purple sweet potato (Ipomea batatas L) contains some bioactive substances, such as anthocyanin and tannin, which help to prevent the prolongation of the inflammatory phase and increase fibroblast proliferation. The aim of this research was to determine the effect of purple sweet potato extract gel toward the expression of TNF-α and skin collagen density on rats with open wound model. The open wound was made circular using a modified Morton method. This study used a completely randomized design with 20 rats, Wistar strain aged 12 weeks, weighing 150-200 g, divided into 5 groups with 4 replicates. TNF-α expression observed in immunohistochemistry method and analyzed using one-way ANOVA statistical test, followed by Honestly Significant Difference Test. The density of collagen with Masson Trichome staining was analyzed by the Kruskal-Wallis nonparametric statistic test with Mann-Whitney-U test. The results showed that the purple sweet potato extract as a therapy for open wounds was significantly reduce the TNF-α expression and increase collagen density. The conclusion is that the purple sweet potato extract with 30% concentration is the most effective therapy in increasing collagen density but not very effective in reducing TNF-α expression.

Analisis Jumlah Sel Monosit yang Mengekspresikan Tnf-Α setelah dipapar Porphyromonas Gingivalis dan Tulang Ikan Kuniran (Upeneus Sulphureus) dengan Teknik Imunositokimia

Monocytes play a role in the first defense against microorganisms that harm the body by activating nuclear factor kβ (Nf-kβ) which is the main regulator of inflammatory genes, one of which is TNF-α. TNF-α at excessive levels will cause tissue damage. Therefore an immunomodulator is needed which can regulate the body's immune system. One natural ingredient that acts as an immunomodulator is fish bones. Purpose of research: To analyze the effect of goatfish’s bone extract on TNF-α expression in human monocytes exposed to P. gingivalis. Method: experimental laboratories in vitro. TNF-α expression was tested by imunocytochemical techniques. Results: The results showed that the average number of monocytes expressing TNF-α in group P1 (group of diclofenac sodium) was 42.33 cells per cell. In the P2 group (P. gingivalis group) was 95 cell percent cells and for P3 (fish bone group) was 74.33 cells per cell and the control group was 11.66 cells per hundred cells. Conclusion: goatfish’s bone extract can act as an anti-inflammatory which is effective in reducing TNF-α expression in monocyte exposed to P. gingivalis but less effective than diclofenac sodium.

The potency of eel meat (Monopterus albus) extract on tumor necrosis factor alpha (TNF-α) expression and epidermal thickness incision wounds on rats (Rattus norvegicus).

Objective:The objective of this study is to determine the effect of eel meat (Monopterus albus) extract ointment on an incision.Materials and Methods:The experimental animals used in this study were 20 male rats (Rattus norvegicus), Wistar, weighing 150–200 gm and aged 8–12 weeks. This study uses complete random design and is divided into four groups. In the negative control group, group treatment was carried out on the healthy rat. In the positive control group, the incision was performed without any therapy. In the T1 and T2 groups, group treatment was performed with a dose of 2% and 5% eel (M. albus) extracts. The TNF-α expression was analyzed by the immunohistochemistry (IHC) technique and epidermal thickness by Masson’s Trichrome (MT) staining. Data analysis of TNF-α expression and epidermal thickness was done using one-way analysis of variance and the Tukey test with a confidence level of 95% (α = 0.05).Results:The results showed that the eel (M. albus) extract therapy, with a concentration of 2% and 5%, significantly (p < 0.05) reduced the TNF-α expression and increased the epidermal thickness.Conclusion:It can be concluded that the administration of eel (M. albus) extract therapy could help to reduce TNF-α expression and increase epidermal thickness in rat incision wounds.

The Protein Kinase C Inhibitor MS-553 for the Treatment of Chronic Lymphocytic Leukemia

The Protein Kinase C Inhibitor MS-553 for the Treatment of Chronic Lymphocytic Leukemia

Effects of cyclophosphamide combined with prednisone on TNF-α expression in treatment of patients with interstitial lung disease.

Effects of cyclophosphamide combined with prednisone on TNF-α expression in the treatment of patients with interstitial lung disease (ILD), and its clinical significance were investigated. A prospective analysis was performed on 198 patients with ILD in Jinan Central Hospital Affiliated to Shandong University from January 2010 to December 2017. Among them, 101 patients treated with cyclophosphamide combined with prednisone were assigned in the combined treatment group, and 97 patients treated with prednisone alone in the control group. Patients in the two groups were compared in terms of lung function, St. George's Respiratory Questionnaire (SGRQ) score, clinical efficacy, adverse reactions and TNF-α expression levels before and after treatment. After treatment, the patients in the combined treatment group had significantly higher forced vital capacity (FVC) and forced expiratory volume in first second (FEV1) compared with the control group, but significantly lower diffusing capacity of lung for carbon monoxide (DLCO) and DLCO% (P<0.05). In both groups, patients after treatment had higher FVC and FEV1, but lower DLCO and DLCO% (P<0.05), compared with before treatment, while SGRQ score before treatment was higher than that after treatment (P<0.05). Compared with control group, the combined treatment group had significantly more patients with complete remission (CR) and higher total effective rate, however less patients with stable disease (SD) (P<0.05). Patients with adverse reactions in the combined treatment group were less than those in the control group (P<0.05). After treatment, TNF-α expression level in the combined treatment group was significantly lower than that in the control group (P<0.05), and TNF-α expression before treatment was higher than that after treatment in both groups (P<0.05). In conclusion, cyclophosphamide combined with prednisone is effective and safe in the treatment of ILD without severe adverse reactions, reducing TNF-α expression level, and therefore is worthy of clinical application.

P2583Endothelial cell-derived extracellular vesicles promote M2 polarization of THP-1 cells

Abstract Introduction The intercellular transfer of biologically active molecules in extracellular vesicles (EVs) has recently been discovered as an important mechanism, which regulates cardiovascular health and disease. In the context of atherosclerosis, endothelial cell-derived EVs have been shown to modulate the phenotype of EV recipient cells in a relevant manner. Under pro-atherogenic conditions e.g. hyperglycemia, the export of numerous signal molecules in EVs is altered and so are EV-dependent effects on recipient cells. While the effect of endothelial-cell derived EVs on other endothelial cells and vascular smooth muscle cells is well characterized, little is known about the vesicle-based interaction of endothelial cells and monocytes under pro-atherogenic conditions. This is particularly relevant as monocytes are crucial modulators of vascular regeneration and inflammation. Our aim was therefore to investigate, whether EVs from endothelial cells have a significant effect on the differentiation and polarization of monocytes and how this process is affected by pathologic conditions as mentioned above. Methods and results Human Coronary Arterial Endothelial Cells (HCAECs) were cultured in high-Glucose-medium (30 mM) for 72h. PBS was used as a control. Large EVs were isolated from the culture supernatant by differential centrifugation (1 x 1500 g / 15 min + 2 x 2ehz748.0909 g / 40 min). The harvested large EVs were characterized by Immunoblotting, Nanoparticle Tracking Analysis as well as Transmission electron microscopy and were shown to be mostly between 80 and 500 nm in size. Specific surface markers including Annexin V and Flotillin-1 were highly enriched in the isolated EVs. The EVs were used for co-culture with THP-1 cells with and without previous phorbol-12-myristate-13-acetate (PMA) stimulation. After 4h as well as after 24 h of incubation with EVS, total RNA was isolated from the THP-1 cells and qPCR was performed to assess polarization towards M1 by TNF-α gene expression or M2 by IL-10 expression. While EV treatment of THP-1 cells without previous PMA-stimulation showed no measurable effect, a significant decrease in the expression of TNF-α was detected after 4 h of treatment from Glucose injured HCAECs. Similar results were obtained without glucose stimulation, the most significant reduction of TNF-α expression however was seen at 24 h. In regard to IL-10 no significant expression changes were detected in EV treated THP-1 cells. Conclusion We showed that glucose injury does not relevantly affect vesicle release or size. Furthermore, endothelial cell derived EVs cause a reduction of TNF-α expression, which indicates a polarization towards an M2 macrophage phenotype, irrespective of prior hyperglycaemic injury. As the M2 phenotype has been described as pro-regenerative, we conclude that endothelial cell derived EVs can exert a protective function during the invasion of monocytes in cardiovascular disease and remodeling.

Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 Alleviate Bacterial Vaginosis and Osteoporosis in Mice by Suppressing NF-κB-Linked TNF-α Expression.

Excessive expression of TNF-α worsens bacterial vaginosis (BV) and osteoporosis. Therefore, to understand whether probiotics could alleviate vaginosis and osteoporosis, we isolated anti-inflammatory Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 from kimchi and human fecal lactic acid bacteria collection and examined their effects on Gardnerella vaginalis (GV)-induced vaginosis and ovariectomy-induced osteoporosis in female mice. Oral gavage of NK3 and/or NK49 significantly alleviated GV-induced vaginosis; these inhibited NF-κB activation and TNF-α expression in the vagina and uterus, and decreased the GV population in the vagina. Furthermore, treatment with NK3 and/or NK49 alleviated ovariectomy-induced osteoporosis and obesity; these increased blood calcium, phosphorus, and osteocalcin levels and suppressed body weight. GV-induced vaginosis and ovariectomy increased colonic myeloperoxidase activity, TNF-α expression, and fecal Proteobacteria population. NK3 and/or NK49 treatments reduced TNF-α expression and NF-κB activation in the colon. NK3 and NK49 treatment also restored GV- or ovariectomy-disrupted gut microbiota composition. In conclusion, NK3 and NK49 may simultaneously alleviate BV and osteoporosis by suppressing NF-κB-linked TNF-α expression through the regulation of gut microbiota population.

Accumbens neuroimmune signaling and dysregulation of astrocytic glutamate transport underlie conditioned nicotine-seeking behavior.

Nicotine self-administration is associated with decreased expression of the glial glutamate transporter (GLT-1) and the cystine-glutamate exchange protein xCT within the nucleus accumbens core (NAcore). N-acetylcysteine (NAC) has been shown to restore these proteins in a rodent model of drug addiction and relapse. However, the specific molecular mechanisms driving its inhibitory effects on cue-induced nicotine reinstatement are unknown. Here, we confirm that extinction of nicotine-seeking behavior is associated with impaired NAcore GLT-1 function and expression and demonstrates that reinstatement of nicotine seeking rapidly enhances membrane fraction GLT-1 expression. Extinction and cue-induced reinstatement of nicotine seeking was also associated with increased tumor necrosis factor alpha (TNFα) and decreased glial fibrillary acidic protein (GFAP) expression in the NAcore. NAC treatment (100 mg/kg/day, i.p., for 5 d) inhibited cue-induced nicotine seeking and suppressed AMPA to NMDA current ratios, suggesting that NAC reduces NAcore postsynaptic excitability. In separate experiments, rats received NAC and an antisense vivo-morpholino to selectively suppress GLT-1 expression in the NAcore during extinction and were subsequently tested for cue-induced reinstatement of nicotine seeking. NAC treatment rescued NAcore GLT-1 expression and attenuated cue-induced nicotine seeking, which was blocked by GLT-1 antisense. NAC also reduced TNFα expression in the NAcore. Viral manipulation of the NF-κB pathway, which is downstream of TNFα, revealed that cue-induced nicotine seeking is regulated by NF-κB pathway signaling in the NAcore independent of GLT-1 expression. Ultimately, these results are the first to show that immunomodulatory mechanisms may regulate known nicotine-induced alterations in glutamatergic plasticity that mediate cue-induced nicotine-seeking behavior.

Neuregulin-1 Fosters Supportive Interactions between Microglia and Neural Stem/Progenitor Cells.

Microglia play diverse roles in homeostasis and pathology of the central nervous system (CNS). Their response to injury or insult is critical for initiating neuroinflammation and tissue damage as well as resolution of inflammation and wound healing. Changes to the microenvironment of microglia appear to be a key determinant of their phenotype and their role in the endogenous repair process in the injured or diseased CNS. Our recent findings have identified a positive role for neuregulin-1 (Nrg-1) in regulating immune response in spinal cord injury and focal demyelinating lesions. We show that increasing the tissue availability of Nrg-1 after injury can promote endogenous repair by modulating neuroinflammation. In the present study, we sought to elucidate the specific role of Nrg-1 in regulating microglial activity and more importantly their influence on the behavior of neural stem/progenitor cells (NPCs). Using injury-relevant in vitro systems, we demonstrate that Nrg-1 attenuates the expression of proinflammatory mediators in activated microglia. Moreover, we provide novel evidence that availability of Nrg-1 can restore the otherwise suppressed phagocytic ability of proinflammatory microglia. Interestingly, the presence of Nrg-1 in the microenvironment of proinflammatory microglia mitigates their inhibitory effects on NPC proliferation. Nrg-1 treated proinflammatory microglia also augment mobilization of NPCs, while they had no influence on their suppressive effects on NPC differentiation. Mechanistically, we show that Nrg-1 enhances the interactions of proinflammatory microglia and NPCs, at least in part, through reduction of TNF-α expression in microglia. These findings provide new insights into the endogenous regulation of microglia-NPC interactions and identify new potential targets for optimizing this important crosstalk during the regenerative process after CNS injury and neuroinflammatory conditions.

The Benzimidazole Derivatives, B1 (N-[(1H-Benzimidazol-2-yl)Methyl]-4-Methoxyaniline) and B8 (N-{4-[(1H-Benzimidazol-2-yl)Methoxy]Phenyl}Acetamide) Attenuate Morphine-Induced Paradoxical Pain in Mice.

Despite being routinely used for pain management, opioid use is limited due to adverse effects such as development of tolerance and paradoxical pain, including thermal hyperalgesia and mechanical allodynia. Evidence indicates that continued morphine administration causes increased expression of proinflammatory mediators such as tumor necrosis factor-alpha (TNF-α). The objectives of the present study were to determine the effects of B1 (N-[(1H-benzimidazol-2-yl)methyl]-4-methoxyaniline) and B8 (N-{4-[(1H-benzimidazol-2-yl)methoxy]phenyl}acetamide), benzimidazole derivatives, on thermal nociception and mechanical allodynia during repeated morphine (intraperitoneal; 5 mg/kg twice daily for 6 days)-induced paradoxical pain and TNF-α expression in the spinal cord in mice. Our data indicate that administration of benzimidazole derivatives attenuated morphine-induced thermal hyperalgesia and mechanical allodynia. Benzimidazole derivatives also reduced TNF-α expression in mice. Taken together, these results suggest that benzimidazole derivatives might be useful for the treatment of neuroinflammatory consequences of continued morphine administration and could be potential drug candidates for the management of opioid-induced paradoxical pain.

A novel antimicrobial peptide-derived vehicle for oligodeoxynucleotide delivery to inhibit TNF-α expression

Indolicidin (IL), an antimicrobial peptide, was investigated as a vehicle to promote oligodeoxynucleotides (ODNs) delivery. To increase charge density, IL was dimerized by adding a cysteine to its C or N terminus, which was denoted as ILC or CIL, respectively. In contrast to IL, cytotoxicity of ILC and CIL was significantly reduced because these dimeric peptides were longer than IL, which restricted their insertions to cell membrane. In contrast to ILC, CIL displayed well loading efficiency. These peptides were applied to deliver ODNs against tumor necrosis factor-α (TNF-α) because TNF-α is a pro-inflammatory cytokine which plays an important role in immunological diseases. Although IL/ODN slightly reduced TNF-α expression, the high cytotoxicity restricted its application window. Furthermore, ILC/ODN was incapable of inducing gene silence due to its low encapsulation efficiency and poor endosomal escape. In contrast, CIL exhibited excellent ODN transportation and the internalized CIL/ODN complexes may escape from endosomes. Therefore, TNF-α expression can be specifically reduced by CIL/ODN complexes, and the silence effect was maintained longer than 14 h. This study provides a useful strategy of peptide vehicle design, which may facilitate the delivery of not only ODN but also other oligonucleotides, including siRNA and miRNA, to promote gene silence application.

Haptoglobin improves acute phase response and endotoxin tolerance in response to bacterial LPS

Sepsis is characterized by delayed acute phase response and lowered immune tolerance in patients. Acute phase serum proteins, like Haptoglobin (Hp), have been associated with increased mortality in bacteria mediated acute lung inflammation and sepsis in neonates. However, it’s direct role in modulating the immune response by regulating pro-inflammatory mediators leading to immune tolerant state and if gender affects its expression levels during bacterial infection, especially in blood has not been fully explored. To understand its specific role in endotoxin-mediated immune response, we investigated the correlation between the rise in Hp levels on bacterial infection and its influence on the expression of pro-inflammatory mediators in male and female Whole blood (WHB) and PBMCs. Here, we observed pathogen-specific and gender-specific expression of Hp. Gonadal steroid hormones differentially influenced the Hp expression in LPS-induced WHB, where the addition of Estrogen increased Hp expression, with suppression of TNFα, in both genders. Further on evaluating, the influence of Hp on TNFα expression in endotoxin tolerance (ET), we show that increased Hp levels directly reduced TNFα expression in ET models. Interestingly, blockade of secreted Hp significantly reversed the (ET) state, confirmed by a significant rise in TNFα expression in both ex vivo and in vitro ET models, indicating a possible feedback inhibition by Hp on inflammatory mediators like TNFα. We also investigated the role of PKCδ in the regulation of LPS induced secretion of acute phase proteins (Hp) in serum, where inhibition of PKCδ, reduced secretion of anti-microbial proteins in response to LPS shown by restored bacterial growth. These findings clearly highlight the crucial role of Hp in maintaining immune tolerance via suppressing the pro-inflammatory mediators and also in preventing bacterial proliferation in blood during infection.

Ac2-26 Induces IKKβ Degradation Through Chaperone-Mediated Autophagy Via HSPB1 in NCM-Treated Microglia.

Annexin A1 (ANXA1) is an endogenous protein with potent anti-inflammatory properties in the brain. Although ANXA1 has been predominantly studied for its binding to formyl peptide receptors (FPRs) on plasma membranes, little is known regarding whether this protein has an anti-inflammatory effect in the cytosol. Here, we investigated the mechanism by which the ANXA1 peptide Ac2-26 decreases high TNF-α production and IKKβ activity, which was caused by oxygen glucose deprivation/reperfusion (OGD/R)-induced neuronal conditioned medium (NCM) in microglia. We found that exogenous Ac2-26 crosses into the cytoplasm of microglia and inhibits both gene expression and protein secretion of TNF-α. Ac2-26 also causes a decrease in IKKβ protein but not IKKβ mRNA, and this effect is inverted by lysosome inhibitor NH4CL. Furthermore, we demonstrate that Ac2-26 induces IKKβ accumulation in lysosomes and that lysosomal-associated membrane protein 2A (LAMP-2A), not LC-3, is enhanced in microglia exposed to Ac2-26. We hypothesize that Ac2-26 mediates IKKβ degradation in lysosomes through chaperone-mediated autophagy (CMA). Interestingly, ANXA1 in the cytoplasm does not interact with IKKβ but with HSPB1, and Ac2-26 promotes HSPB1 binding to IKKβ. Furthermore, both ANXA1 and HSPB1 can interact with Hsc70 and LAMP-2A, but IKKβ only associates with LAMP-2A. Downregulation of HSPB1 or LAMP-2A reverses the degradation of IKKβ induced by Ac2-26. Taken together, these findings define an essential role of exogenous Ac2-26 in microglia and demonstrate that Ac2-26 is associated with HSPB1 and promotes HSPB1 binding to IKKβ, which is degraded by CMA, thereby reducing TNF-α expression.

The effect of purple rice (Oryza sativa L. indica) extract on the inflammatory response in a colon cancer cell line and dextran sulfate-induced tumor promotion in the rat colon

This study investigated the effects of purple rice extract (PRE ) on inflammation-related colon carcinogenesis. Administration of PRE 1 g/kg bodyweight reduced both the number (39.05%) and size of aberrant crypt foci (ACF) (14.28%) in the colons of rats that received dimethylhydrazine (DMH) initiation with promotion by dextran sulfate sodium (DSS) (P<0.05) with the reduction of TNF-α expression in colonic epithelial cells. On the other hand, PRE could suppress the expression of both TNF-α and IL-6 in RAW 264.7 cells activated by lipopolysaccharide (LPS) at both the mRNA and protein levels. Moreover it could reduce the expression of TNF-α and IL-1β mRNA in inflamed HT-29. These findings suggested that PRE is able to modulate the inflammatory process in both activated macrophages and colon cancer cells. Therefore, PRE suppressed macrophage activation and colon cancer cell line response to inflammation and modulated of inflammation induced ACF progression in the rat colon.

Hyaluronic acid 35 normalizes TLR4 signaling in Kupffer cells from ethanol-fed rats via regulation of microRNA291b and its target Tollip

TLR4 signaling in hepatic macrophages is increased after chronic ethanol feeding. Treatment of hepatic macrophages after chronic ethanol feeding with small-specific sized hyaluronic acid 35 (HA35) normalizes TLR4 signaling; however, the mechanisms for HA35 action are not completely understood. Here we used Next Generation Sequencing of microRNAs to identify negative regulators of TLR4 signaling reciprocally modulated by ethanol and HA35 in hepatic macrophages. Eleven microRNAs were up-regulated by ethanol; only 4 microRNAs, including miR291b, were decreased by HA35. Bioinformatics analysis identified Tollip, a negative regulator of TLR4, as a target of miR291b. Tollip expression was decreased in hepatic macrophages from ethanol-fed rats, but treatment with HA35 or transfection with a miR291b hairpin inhibitor restored Tollip expression and normalized TLR4-stimulated TNFα expression. In peripheral blood monocytes isolated from patients with alcoholic hepatitis, expression of TNFα mRNA was robustly increased in response to challenge with lipopolysaccharide. Importantly, pre-treatment with HA35 reduced TNFα expression by more than 50%. Taken together, we have identified miR291b as a critical miRNA up-regulated by ethanol. Normalization of the miR291b → Tollip pathway by HA35 ameliorated ethanol-induced sensitization of TLR4 signaling in macrophages/monocytes, suggesting that HA35 may be a novel therapeutic agent in the treatment of ALD.

CD39 and CD73 activity are protective in a mouse model of antiphospholipid antibody-induced miscarriages

ObjectiveAntiphospholipid syndrome (APS) is a systemic autoimmune disorder of young adults associated with devastating pregnancy complications (recurrent miscarriages, preeclampsia and low birth weight) and vascular complications including thrombosis. The key components implicated in pathogenesis of APS are the complement cascade and tissue factor (TF) activity causing inflammation and coagulation. Purinergic signalling involving catabolism of ATP to adenosine by cell-surface enzymes CD39 and CD73 has anti-inflammatory and anti-thrombotic effects. We studied whether activities of CD39 and CD73 are important in preventing the development of miscarriages in APS. MethodsWe studied frequency of miscarriages and decidual pathology following passive transfer of human aPL-ab to pregnant wildtype mice, and mice deficient in CD39 and CD73, and also transgenic mice exhibiting 2-3X higher CD39 activity. ResultsaPL-ab infusion in pregnant CD39-or CD73-knockout mice triggers an increase in miscarriages, associated with increased TF expression and complement deposition as well as elevated oxidative stress and pro-inflammatory TNF-α and IL-10 expression within the placental decidua. In contrast, aPL-ab induced miscarriages are prevented in mice over-expressing CD39, with reduced decidual TF expression and C3d deposition, diminished lipid peroxidation (4-hydroxynonenal or 4-HNE positive lipid adducts), and reduced TNF-α expression. ConclusionWe demonstrate a protective role for CD39 in APS and provide rationale for both the development of endothelial cell-targeted soluble CD39 as a novel therapeutic for APS and analysis of perturbations in the purinergic pathway to explain human disease.

Association of the gene expression variation of tumor necrosis factor-α and expressions changes of dopamine receptor genes in progression of diabetic severe foot ulcers.

Objective(s):Regulation of pro-inflammatory factors such as TNF-α which are secreted by the immune cells through induction of their several receptors including dopamine receptors (especially DRD2 and DRD3) is one of the noticeable problems in diabetic severe foot ulcer healing. This study was conducted to evaluate the alteration of TNF-αin plasma as well as DRD2 and DRD3 changes in PBMCs of diabetics with severe foot ulcers.Materials and Methods:Peripheral blood samples were collected from 31 subjects with ulcers, 29 without ulcers, and 25 healthy individuals. Total mRNA was extracted from PBMCs for the study of DRD2, DRD3, and TNF-α gene expression variations. Expression patterns of these genes were evaluated by real-time PCR. Consequently, concentration of TNF-α was investigated in plasma.Results:Significant decrease in gene expression and plasma concentration of TNF-α in PBMCs was observed in both patient groups at P<0.05. These diminutions are correlated to the decrease in the expression of both DRD2 and DRD3 in PBMCs of both patient groups. Also, the same relationship is present between expressions of two new DRD3 transcripts with TNF-α downturn.Conclusion:We concluded that DRD2 and DRD3 expression alteration and presence of new DRD3 transcripts can be effective in reduction of TNF-α expression as a pro-inflammatory factor. Performing complementary studies, may explain that variations in DRD2 and DRD3 are prognostic and effective markers attributed to the development of diabetes severe foot ulcers.

The inhibition of 5-Lipoxygenase (5-LO) products leukotriene B4 (LTB4) and cysteinyl leukotrienes (cysLTs) modulates the inflammatory response and improves cutaneous wound healing

To analyze the participation of the enzyme 5-lipoxygenase (5-LO) in skin repair, WT wounds were compared to those in 5-LO deficient mice (5-LO−/−), which presented faster closure and reduced inflammatory infiltrate in the skin, together with increased CD4 regulatory T cells markers in the draining lymph nodes. The 5-LO−/− wounds also had diminished TNF-α, CCL11, CCL7, CCL2, CXCL9, CCR1 and CXCR2 mRNA expression in the lesions, besides differential extracellular matrix remodeling. Furthermore, when cysteinyl leukotriene (cysLT) and leukotriene (LTB4) receptors were antagonized in WT mice, there was a remarkable reduction in TNF-α expression and faster skin healing, similarly to the findings in 5-LO−/− animals. Finally, our results suggested that 5-LO products, in special cysLT and LTB4, underline skin inflammation that follows skin injury and their neutralization may be an important strategy to improve cutaneous healing.