Decreased TNF-α Expression Research Articles

Protective effects of propolis extract against nicotine-evoked pulmonary and hepatic damage.

There is increasing interest in the use of natural products to treat many diseases, considering the minimal toxicity, availability, and low cost. Propolis, a natural resinous product produced by honeybees, has been proven for its antioxidant and anti-inflammatory properties. Therefore, this study was designed to investigate the protective potential of propolis extract against nicotine-induced pulmonary and hepatic damage in rats. Sprague Dawley rats were divided into six groups: control, propolis (200 and 300 mg/kg, p.o.), nicotine (10 mg/kg, i.p), and nicotine plus propolis-treated groups. Nicotine and propolis were given every day for 8 weeks. Then, blood and bronchoalveolar lavage fluid (BALF) were collected for assessing liver and lung functions. Liver and lung tissues were also harvested to assess oxidative stress and inflammatory biomarkers in addition to histopathological and immunohistochemical analysis. Both doses of propolis significantly decreased AST, ALT, ALP, and total and differential cell counts in a dose-dependent manner. Propolis extract significantly attenuated oxidative stress in both lung and liver tissues. The restoration of antioxidant status (GSH level, SOD activities) and reduction of nitric oxide and MDA content was more so in propolis 300-treated than propolis 200-treated group. This was parallel to the improvement seen in histopathological examination. Propolis 200 and 300 significantly decreased Nrf2 expression and increased HO-1 expression in a dose-dependent manner. Moreover, immunohistochemical examination revealed that propolis 200 and 300 decreased the expression of iNOS in lung and liver tissues while decreased TNF-α expression in lung tissues only. Propolis extract could have a protective potential against nicotine-induced pulmonary and hepatic damage via activating Nrf2/HO-1 signaling.

Biocompatible valproic acid-coupled nanoparticles attenuate lipopolysaccharide-induced inflammation

Inflammatory diseases like sepsis are associated with dysregulated gene expression, often caused by an imbalance of epigenetic regulators, such as histone acetyltransferases (HATs) and histone deacetylases (HDACs), and consequently, altered epigenetic chromatin signatures or aberrant posttranslational modifications of signalling proteins and transcription factors. Thus, HDAC inhibitors (HDACi) are a promising class of anti-inflammatory drugs. Recently, an efficient drug delivery system carrying the class I/IIa selective HDACi valproic acid (VPA) was developed to circumvent common disadvantages of free drug administration, e.g. short half-life and side effects. The cellulose-based sulphated VPA-coupled (CV-S) nanoparticles (NPs) are rapidly taken up by cells, do not cause any toxic effects and are fully biocompatible. Importantly, VPA is intracellularly cleaved from the NPs and HDACi activity could be proven. Here, we demonstrate that CV-S NPs exhibit overall anti-inflammatory effects in primary human macrophages and are able to attenuate the lipopolysaccharide-induced inflammatory response. CV-S NPs show superior potential to free VPA to suppress the TLR-MyD88-NF-κB signalling axis, leading to decreased TNF-α expression and secretion.

Involvement of the Microglial Aryl Hydrocarbon Receptor in Neuroinflammation and Vasogenic Edema after Ischemic Stroke.

Microglia are activated after ischemic stroke and induce neuroinflammation. The expression of the aryl hydrocarbon receptor (AhR) has recently been reported to elicit cytokine expression. We previously reported that microglial activation mediates ischemic edema progression. Thus, the purpose of this study was to examine the role of AhR in inflammation and edema after ischemia using a mouse middle cerebral artery occlusion (MCAO) model. MCAO upregulated AhR expression in microglia during ischemia. MCAO increased the expression of tumor necrosis factor α (TNFα) and then induced edema progression, and worsened the modified neurological severity scores, with these being suppressed by administration of an AhR antagonist, CH223191. In THP-1 macrophages, the NADPH oxidase (NOX) subunit p47phox was significantly increased by AhR ligands, especially under inflammatory conditions. Suppression of NOX activity by apocynin or elimination of superoxide by superoxide dismutase decreased TNFα expression, which was induced by the AhR ligand. AhR ligands also elicited p47phox expression in mouse primary microglia. Thus, p47phox may be important in oxidative stress and subsequent inflammation. In MCAO model mice, P47phox expression was upregulated in microglia by ischemia. Lipid peroxidation induced by MCAO was suppressed by CH223191. Taken together, these findings suggest that AhR in the microglia is involved in neuroinflammation and subsequent edema, after MCAO via p47phox expression upregulation and oxidative stress.

The Effect of Azadirachta indica leaves extract on Transforming Growth Factor-β and Tumor Necrosis Factor-α in a Plasmodium berghei infected Mice model

Malaria is a health problem for the world's population and is predominantly located in tropical and subtropical areas. The three countries with the most malaria cases are India (58%), followed by Indonesia (20%), then Myanmar (16%). This study aims to determine the effect of neem leaf extract on increasing TGF-β expression and decreasing TNF-α expression in mice infected with Plasmodium berghei. In this study there were four groups, namely Treatment 1 (in Plasmodium berghei infection without therapy). Treatment 2 (in Plasmodium berghei infection and treated with Azadirachta indica leaf extract at a dose of 0.25 mg / g BW). Treatment 3 = (in Plasmodium berghei infection and therapy with Azadirachta indica leaves at a dose of 0.5 mg / g BW). Treatment 4 = (in Plasmodium berghei infection and treated with Azadirachta indica leaves at a dose of 1 mg / g BW). TGF-β examination by elisa method and TNF-α by immunohistochemistry. Data analysis using SEM (Structural Equation Modeling) The results of treatment 1 and 2 showed a decrease in plasma TGF-β expression (t = 1.13; tcount = 1.93; ≥ttabel = 1.96) and spleen (tcount = 1.53; tcount = 1.45; ≥ttabel = 1.96) but there was an increase in spleen TNF-α expression (tcount = 1.77; tcount = 1.00; ≥ttabel = 1.96). Groups 3 and 4 showed an increase in plasma TGF-β expression (tcount = 5.13; tcount = 2.42; ≥ttable = 1.96) and spleen (tcount = 2.00; tcount = 1.97; ≥ttabel = 1.96) but there was a decrease in spleen TNF-α expression (tcount = 2.03; tcount = 2.11; ≥ttabel = 1.96). Conclusion: Azadirachta indica leaf therapy can increase TGF-β expression and decrease TNF-α expression in the spleen.
 Keywords: Azadirachta indica ethanol extract, transforming growth factor- β, tumor necrosis factor-α

Abstract 419: PKD1 regulates susceptibility to ulcerative colitis and colorectal cancer

Abstract Compromised barrier function of colon epithelial tissue is associated with inflammatory bowel diseases (IBDs), and increases the risk of colorectal cancer (CRC) through a process of tumor-elicited inflammation (TEI). Apical tight junction (TJ) proteins, including ZO-1, occludin and specific claudins, are critical in the maintenance of epithelial barrier function and control of paracellular permeability. Changes in TJ composition in the kidney are strongly linked to the etiology of autosomal dominant polycystic kidney disease (ADPKD), with ADPKD-inducing mutations in PKD1 causing non-leaky barriers that can withstand high hydrostatic pressure within renal cysts a hallmark of this common (~1 in 600) inherited disease. Intriguingly, a large population study suggested decreased incidence of CRC in ADPKD patients, while other studies implicated PKD1 in control of epithelial-mesenchymal transition (EMT) and invasion. Based on these data, we hypothesized that loss of PKD1 might hinder susceptibility to ulcerative colitis and CRC, based on reorganization of TJs and limitation of TEI in the colon. We directly assessed PKD1 control of epithelial barrier function using conditional Pkd1fl/fl mice in which tamoxifen induces Cre-lox dependent targeted inactivation of Pkd1 gene in the colon. We compared 10-12 week old wt or induced Pkd1fl/fl mice treated for 5 days with 2.5% DSS in drinking water to induce colitis, to a non-DSS control group. For wt mice, DSS increased orally gavaged FITC-dextran in the serum versus vehicle-treated mice, and histopathological assessment indicated significant damage to colon. In contrast, DSS did not increase FITC-dextran in the serum of Pkd1−/- mice, and less DSS-induced tissue damage was observed, suggesting reinforced colon barrier function. Further, based on immunofluorescence (IF) analysis of tissue sections, the claudins CLDN4 and CLDN7, associated with barrier impermeability, were strongly elevated in the colonic epithelium of Pkd1fl/fl versus wt mice, with consistently greater localization to cell junctions in Pkd1fl/fl versus wt mice. Using a Cdx2-ERT2/Cre model for tamoxifen-induced loss of Apc and/or Pkd1 in the colon, we compared tumorigenesis in the Apcfl/fl, Apcfl/fl Pkd1fl/fl, and Apcfl/flPkd1fl/+ genotypes. There was a highly significant increase in the number and size of Apcfl/fl-induced tumors based on retention of Pkd1. Loss of Pkd1 substantially increased CLDN4 in normal and tumor tissue, and decreased TNFα expression in tumors, suggesting reduced inflammation. Analysis in organoids recapitulated observed effects of Pkd1, indicating cell-autonomous effect. Epistasis experiments suggest activation of the tumor suppressor CFTR as a mechanism by which loss of PKD1 limits EMT and tumor growth; we found chemical activators of CFTR selectively limited the growth of PKD1-positive organoids. Better understanding of the role of PKD1 and its effectors may suggest therapeutic strategies, and inform genetic counseling, for IBD and CRC. Citation Format: Anna S. Nikonova, Anna A. Kiseleva, Alexander Deneka, Rossella Tricarico, Sergei Grivennikov, Erica Golemis. PKD1 regulates susceptibility to ulcerative colitis and colorectal cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 419.

Effects of dexmedetomidine combined with targeted temperature management on the expression of hippocampal P2X7 receptor and antisense hypoxia inducible factor-1α in rats with traumatic brain injury

Objective To investigate the effects of dexmedetomidine (Dex) combined with targeted temperature management (TTM) on the expression of hippocampal P2X7 receptor and antisense hypoxia inducible factor (aHIF)-1α in rats with traumatic brain injury (TBI). Methods A model of TBI was established through controlled cortical impact using rats after 1 week of adaptive feeding. Then, 15 rats were randomly selected as a control group, while the others were set as a model group. The successfully modeled rats were divided into five groups (n=15), according to the random number table method: a sham operation group, a TBI group, a TBI+Dex group, a TBI+TTM group and a TBI+Dex+TTM group. After corresponding treatment, their behaviors were tested, and the expression of tumor necrosis factor (TNF)-α/interleukin (IL)-1β in brain tissue and the expression of hippocampal P2X7 receptor and aHIF- 1α were analyzed. Results Rats in each group did not die. Compared with the TBI, TBI+Dex and TBI+TTM groups, the TBI+Dex+TTM group presented remarkably reduced escape latency on Days 3-5 and shortened exploration duration on Day 6 (P<0.05), as well as decreased expression of TNF-α and IL-1β in brain tissues (P<0.05), reduced expression of hippocampal P2X7 receptor and increased amounts of aHIF- 1α (P< 0.05). Conclusions Dex combined with TTM can effectively decline the concentration of P2X7 receptor, down-regulate the levels of TNF-α and IL-1β, and increase the level of aHIF- 1α, which can be served as a promising therapy for TBI rats. Key words: Brain injuries, traumatic; Dexmedetomidine; Targeted temperature management; P2X7 receptor; Antisense hypoxia inducible factor- 1α

Chronic Alcohol Dysregulates Skeletal Muscle Myogenic Gene Expression after Hind Limb Immobilization in Female Rats.

Alcohol use and aging are risk factors for falls requiring immobilization and leading to skeletal muscle atrophy. Skeletal muscle regeneration is integral to post-immobilization recovery. This study aimed to elucidate the effects of alcohol and ovarian hormone loss on the expression of genes implicated in muscle regeneration. Three-month-old female rats received an ovariectomy or a sham surgery, consumed an alcohol-containing or control diet for 10 weeks, were subjected to unilateral hind limb immobilization for seven days, and finally were allowed a three (3d)- or 14 (14d)-day recovery. Immobilization decreased the quadriceps weight at 3d and 14d, and alcohol decreased the quadriceps weight at 14d in the nonimmobilized hind limb (NI). At 3d, alcohol decreased gene expression of myoblast determination protein (MyoD) in the immobilized hind limb (IMM) and myocyte enhancer factor (Mef)2C and tumor necrosis factor (TNF)α in NI, and ovariectomy increased MyoD and decreased TNFα expression in NI. At 14d, alcohol increased the gene expression of Mef2C, MyoD, TNFα, and transforming growth factor (TFG)β in IMM and decreased monocyte chemoattractant protein (MCP)1 expression in NI; ovariectomy increased TNFα expression in NI, and alcohol and ovariectomy together increased Mef2C expression in NI. Despite increased TGFβ expression, there was no concomitant alcohol-mediated increase in collagen in IMM at 14d. Overall, these data indicate that alcohol dysregulated the post-immobilization alteration in the expression of genes implicated in regeneration. Whether alcohol-mediated molecular changes correspond with post-immobilization functional alterations remains to be determined.

Protective effects of vitamin D against injury in intestinal epithelium.

Vitamin D deficiency is associated with intestinal barrier dysfunction, which contributes to pathogenesis of acute intestinal injury in children. We aim to investigate the effects of vitamin D on intestinal injury in intestinal epithelial cells and organoids. Lipopolysaccharide (LPS) was used to induce injury in intestinal epithelial cells (IEC-18) and organoids, and the effect of vitamin D was assessed. Cell viability was measured and inflammation cytokines TNFα and IL-8 were quantified. FITC-dextran 4kDa (FD4) permeability was measured using Transwell while tight junction markers were assessed by immunofluorescence staining in IEC-18 and intestinal organoids. Data were compared using one-way ANOVA with Bonferroni post-test. IEC-18 viability was decreased by LPS treatment, but was prevented by vitamin D. The upregulation of inflammation was inhibited by vitamin D, which also decreased epithelium permeability. Vitamin D restored tight junction ZO-1 and claudin 2. In addition, vitamin D decreased TNFα expression and prevented the disruption of ZO-1 in injured organoids. Vitamin D rescued epithelial barrier function by improving permeability and restoring tight junctions, leading to decrease inflammation. This study confirms the protective effects of vitamin D, which could be used as a treatment strategy for infants at risk of developing intestinal injury.

Melatonin Modulation of Radiation and Chemotherapeutics-induced Changes on Differentiation of Breast Fibroblasts.

Melatonin exerts oncostatic actions and sensitizes tumor cells to chemotherapeutics or radiation. In our study, we investigated the effects of docetaxel, vinorelbine, and radiation on human breast fibroblasts and its modulation by melatonin. Docetaxel or vinorelbine inhibits proliferation and stimulates the differentiation of breast preadipocytes, by increasing C/EBPα and PPARγ expression and by downregulating tumor necrosis factor α (TNFα), interleukin 6 (IL-6), and IL-11 expression. Radiation inhibits both proliferation and differentiation through the downregulation of C/EBPα and PPARγ and by stimulating TNFα expression. In addition, docetaxel and radiation decrease aromatase activity and expression by decreasing aromatase promoter II and cyclooxygenases 1 and 2 (COX-1 and COX-2) expression. Melatonin potentiates the stimulatory effect of docetaxel and vinorelbine on differentiation and their inhibitory effects on aromatase activity and expression, by increasing the stimulatory effect on C/EBPα and PPARγ expression and the downregulation of antiadipogenic cytokines and COX expression. Melatonin also counteracts the inhibitory effect of radiation on differentiation of preadipocytes, by increasing C/EBPα and PPARγ expression and by decreasing TNFα expression. Melatonin also potentiates the inhibitory effect exerted by radiation on aromatase activity and expression by increasing the downregulation of promoter II, and COX-1 and COX-2 expression. Our findings suggest that melatonin modulates regulatory effects induced by chemotherapeutic drugs or radiation on preadipocytes, which makes it a promising adjuvant for chemotherapy and radiotherapy sensibilization.

Does physical exercise influence in the development of neuroeschistosomiasis?

Neuroschistosomiasis is a severe form of presentation of schistosomiasis in which Schistosoma spp. affects the central nervous system. This is the first study performed to analyze whether there is any relationship between physical effort and the appearance of neuroschistosomiasis, through clinical, molecular and immunological evaluations. An experimental controlled study using 64 male Balb/c inbred mice divided into four groups according to presence or absence of S. mansoni infection and submitted to physical effort or resting was conducted. Thirteen weeks after exercise training, S. mansoni DNA was detected in the brain or spinal cord in about 30% of the infected animals moreover, only S. mansoni-positive samples showed positive labeling for S. mansoni antigens in the brain or spinal cord, with a striking reaction inside the microglia. However, the behavioral tests did not show any clinical symptoms of neuroschistosomiasis in animals submitted to physical effort or in resting. In animals with S. mansoni-positive DNA, immunohistochemical data revealed astrogliosis and microgliosis, elevated IL-10 levels and decreased TNF-α expression. This study demonstrated that isometric exercise does not promote neuroschistosomiasis, furthermore, ectopic forms of schistosomiasis in the central nervous system were largely asymptomatic and exhibited a Th2 immune response profile. More experimental studies are necessary in order to characterize the pathological process of experimental neuroschistosomiasis.

Abstract 3459: PKD1 regulates susceptibility to ulcerative colitis and colorectal cancer

Abstract Compromised integrity of the epithelial barrier function of colon tissue associated with inflammatory bowel diseases (IBDs) or arising from tumor-elicited inflammation (TEI) are associated with and support formation of colorectal cancer (CRC). Apical tight junction (TJ) proteins, including multiple specific claudins, are critical in regulating TJ barrier function and paracellular permeability. Changes in TJ composition in the kidney are strongly linked to the etiology of autosomal dominant polycystic kidney disease (ADPKD), with ADPKD-inducing mutations in PKD1 causing non-leaky barriers that can withstand high hydrostatic pressure within renal cysts a hallmark of this common (1 in 500) inherited disease. Intriguingly, a large population study has found a decreased incidence of CRC in ADPKD patients. Based on these and other suggestive data, we hypothesized that loss of PKD1 would hinder development of ulcerative colitis and initiation of colitis associated cancer, based on reorganizing TJs and TEI in the colon. To evaluate the impact of Pkd1 loss on colon barrier function, we treated 10-12 week old wt or Pkd1fl/fl mice with tamoxifen-induced, Cre expression from the CreERT2 cassette for 5 days with 2.5% DSS in drinking water to induce acute colitis, compared to a no-DSS control group. For the wtcohort, treatment with DSS increased orally gavaged FITC-dextran detectable in serum versus controls. In contrast, no Pkd1-/- mice showed a similar DSS-dependent response to FITC-dextran, suggesting reinforced colon barrier function; further, histopathological assessment confirmed less DSS-induced damage in Pkd1fl/fl mice. Further, based on immunofluorescence (IF) analysis of tissue sections, claudins 4 and 7 associated with increased barrier function) strongly elevated in the colonic epithelium of Pkd1fl/fl versus wt mice, with the expressed proteins having consistently greater localization to cell junctions in Pkd1fl/fl versus wt mice. We also investigated the formation of CRC tumors, using a CDX2-ERT2/Cre model for tamoxifen-induced loss of Apc and/or Pkd1 in the colon to compare tumorigenesis in the Apcfl/fl, Apcfl/fl Pkd1fl/fl, and Apcfl/flPkd1fl/+ genotypes. We observed a highly significant progressive increase in the number and size of Apc mutation-induced tumors based on retention of the Pkd1 gene. Preliminary qRT-PCR analysis of these tumors shows that a Pkd1fl/fl genotype substantially increases CLDN4 in normal and tumor tissue, and decreases TNFα expression in tumors, suggesting reduced inflammation. This work and extended mechanistic characterization identifies PKD1 as a vital regulator of signaling systems already associated with colitis and CRC. Understanding the role of PKD1 and its effectors may provide useful information to genetic counselors assessing risk of IBD and CRC, and suggest therapeutic strategies. Citation Format: Anna S. Nikonova, Anna Kiseleva, Ilya Serebriiskii, Sergei Grivennikov, Erica A. Golemis. PKD1 regulates susceptibility to ulcerative colitis and colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3459.

Effects of Obesity on Pulmonary Inflammation and Remodeling in Experimental Moderate Acute Lung Injury.

Obese patients are at higher risk of developing acute respiratory distress syndrome (ARDS); however, their survival rates are also higher compared to those of similarly ill non-obese patients. We hypothesized that obesity would not only prevent lung inflammation, but also reduce remodeling in moderate endotoxin-induced acute lung injury (ALI). Obesity was induced by early postnatal overfeeding in Wistar rats in which the litter size was reduced to 3 pups/litter (Obese, n = 18); Control animals (n = 18) were obtained from unculled litters. On postnatal day 150, Control, and Obese animals randomly received E. coli lipopolysaccharide (ALI) or saline (SAL) intratracheally. After 24 h, echocardiography, lung function and morphometry, and biological markers in lung tissue were evaluated. Additionally, mediator expression in neutrophils and macrophages obtained from blood and bronchoalveolar lavage fluid (BALF) was analyzed. Compared to Control-SAL animals, Control-ALI rats showed no changes in echocardiographic parameters, increased lung elastance and resistance, higher monocyte phagocytic capacity, collagen fiber content, myeloperoxidase (MPO) activity, and levels of interleukin (IL-6), tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, and type III (PCIII), and I (PCI) procollagen in lung tissue, as well as increased expressions of TNF-α and monocyte chemoattractant protein (MCP)-1 in blood and BALF neutrophils. Monocyte (blood) and macrophage (adipose tissue) phagocytic capacities were lower in Obese-ALI compared to Control-ALI animals, and Obese animals exhibited reduced neutrophil migration compared to Control. Obese-ALI animals, compared to Obese-SAL, exhibited increased interventricular septum thickness (p = 0.003) and posterior wall thickness (p = 0.003) and decreased pulmonary acceleration time to pulmonary ejection time ratio (p = 0.005); no changes in lung mechanics, IL-6, TNF-α, TGF-β, PCIII, and PCI in lung tissue; increased IL-10 levels in lung homogenate (p = 0.007); reduced MCP-1 expression in blood neutrophils (p = 0.009); decreased TNF-α expression in blood (p = 0.02) and BALF (p = 0.008) neutrophils; and increased IL-10 expression in monocytes (p = 0.004). In conclusion, after endotoxin challenge, obese rats showed less deterioration of lung function, secondary to anti-inflammatory and anti-fibrotic effects, as well as changes in neutrophil and monocyte/macrophage phenotype in blood and BALF compared to Control rats.

The neural function repair and mechanism of (-)-epicatechin on murine traumatic brain injury

Objective To investigate whether (-)-epicatechin plays a role in neurological repair on traumatic brain injury in mice. Methods The mice model of traumatic brain injury was established by modified weight drop method. Experimental mice were randomly (random number) divided into the injury+ (Veh) group and the injury + (-)-epicatechin (EC) group. At 3 days after operation, the expression of IL-1β and TNF-α were detected. The number of necrotic cells of the lesion area was detected by PI staining. At 28 days after SCI, Morris Water Maze test was performed to observe the ability of spatial learning and memory in mice. The expression of neurotrophic factors BDNF and NGF were examined by qRT-PCR. The expression of NeuN was detected by immunofluorescence staining. EdU staining was used to observe the neurogenesis in the SGZ region. Results Compared with the Veh group, EC treated group showed better spatial learning and memory ability in time spent in correct quadrant at day 27 and 28 [24 d: (26.333±5.037)% vs (26.583±5.802)%, P=0.938; 25 d (33.300±4.724)% vs (29.767±3.347)%. P=0.166; 26 d: (41.017±7.246)% vs (32.800±8.145)%, P=0.095; 27 d: (48.017±7.424)% vs (35.267±6.748)%, P=0.011; 28 d: (51.617±9.017)% vs (41.116±6.467)%, P=0.043] and in latency to platform at day 27 and 28 [24 d: (62.967±5.494) s vs (63.917±7.027) s, P=0.800; 25 d: (50.533±10.305) s vs (57.217±13.085) s, P=0.349; 26 d: (40.333±10.526) s vs (50.133±11.039) s, P=0.147; 27 d: (28.717±4.137) s vs (44.533±7.181) s, P=0.001; 28 d: (21.950±6.889) s vs (37.567±5.974) s, P=0.002]. There was a decreased expression of IL-1β and TNF-α and increased level of neurotrophic factor BDNF and NGF after EC treatment in EC treatment group, compared to the veh treatment group [IL-1β and TNF-α: (42.690±3.057) ng/mL and (750.167±51.941) ng/mL vs (71.670±4.996) ng/mL and (1 085.167±68.535) ng/mL, P=0.000 6 and 0.003; BDNF and NGF: 0.543±0.033 and 0.334±0.041 vs 0.756±0.088 and 0.514±0.047, P=0.048 and 0.017)]. EC decreased the cell death near injury area (54.833±5.486 vs 74.000±5.323, P=0.031), increased NeuN positive cells (76.667±6.386 vs 42.167±5.237, P=0.002), and increased neurogenesis in SGZ area (12.667±0.760 vs 7.500±1.258, P=0.031). Conclusions (-)-Epicatechin plays an important role in functional recovery after traumatic brain injury in mice. The underlying mechanisms are closely related to inhibited inflammation, enhanced neurotrophic factors and improved neurogenesis. Key words: Traumatic brain injury; (-)-epicatechin; Nerve repair; Brain derived neurotrophic factor; Inflammation factors

Secretomes from metastatic breast cancer cells, enriched for a prognostically unfavorable LCN2 axis, induce anti-inflammatory MSC actions and a tumor-supportive premetastatic lung.

ABSTRACTCancer metastasis is responsible for the clear majority of cancer-related deaths. Survival and expansion of cancer cells at secondary sites requires that these premetastatic microenvironments be primed by primary tumor cells and their secreted factors. Efforts to date have been limited by immune-deficient in vivo models and/or the need for finely-tuned analysis time points that reduce contributions from early-disseminating cancer cells. In this regard, we developed a tumor cell-free syngeneic breast cancer model for characterizing tumor cell secretome-mediated reprogramming of premetastatic tissues. We demonstrate that secretomes from metastatic breast cancer cells differentially regulate the lung and brain, promoting a tumor-supportive lung microenvironment with both elevated CD73 expression and decreased TNFα expression. Using in vitro models of CD73-positive mesenchymal stem cells (MSCs) and macrophages/monocytes, we tested whether MSCs can mediate anti-inflammatory effects of metastatic breast cancer cells. Notably, conditioned media from metastatic Py230 cells reprogrammed the secretomes of MSCs toward an anti-inflammatory state. Mining transcriptome data from Py8119 and Py230 cells revealed a lipocalin 2 (LCN2) axis that is selectively expressed in the metastatic Py230 cells, predicts poor breast cancer patient survival and is elevated in circulating serum of mice chronically treated with conditioned media from Py230 cells. Taken together, these results establish the utility of an immune-competent tumor cell-free model for characterizing the mechanisms of breast cancer cell priming of the premetastatic niche, demonstrate that MSCs can mediate the anti-inflammatory effects of metastatic breast cancer cells and substantiate LCN2 as a promising therapeutic target for blocking breast cancer progression.

Deletion of translin (Tsn) induces robust adiposity and hepatic steatosis without impairing glucose tolerance.

Objective:Translin knockout (KO) mice display robust adiposity. Recent studies indicate that translin and its partner protein, trax, regulate the microRNA and ATM kinase signaling pathways, both of which have been implicated in regulating metabolism. In the course of characterizing the metabolic profile of these mice, we found that they display normal glucose tolerance despite their elevated adiposity. Accordingly, we investigated why translin KO mice display this paradoxical phenotype.Methods:To help distinguish between the metabolic effects of increased adiposity and those of translin deletion per se, we compared three groups: (1) wild-type (WT), (2) translin KO mice on a standard chow diet, and (3) adiposity-matched WT mice that were placed on a high-fat diet until they matched translin KO adiposity levels. All groups were scanned to determine their body composition and tested to evaluate their glucose and insulin tolerance. Plasma, hepatic and adipose tissue samples were collected and used for histological and molecular analyses.Results:Translin KO mice show normal glucose tolerance whereas adiposity-matched WT mice, placed on a high-fat diet, do not. In addition, translin KO mice display prominent hepatic steatosis that is more severe than that of adiposity-matched WT mice. Unlike adiposity-matched WT mice, translin KO mice display three key features that have been shown to reduce susceptibility to insulin resistance: increased accumulation of subcutaneous fat, increased levels of circulating adiponectin and decreased Tnfα expression in hepatic and adipose tissue.Conclusions:The ability of translin KO mice to retain normal glucose tolerance in the face of marked adipose tissue expansion may be due to the three protective factors noted above. Further studies aimed at defining the molecular bases for this combination of protective phenotypes may yield new approaches to limit the adverse metabolic consequences of obesity.

Cellular Behavior of RAW264.7 Cells in 3D Poly(ethylene glycol) Hydrogel Niches.

Although macrophages undergo dynamic cellular responses in diverse extracellular environments, macrophage research has mostly relied on conventional culture methodologies such as two-dimensional and suspension cultures. In contrast, recent efforts have revealed evidence of the characteristic cellular behaviors of macrophages in actual tissues using a three-dimensional (3D) culture matrix. In this work, we exploited a poly(ethylene glycol)-based hydrogel as a macrophage culture matrix and observed cellular behaviors in 3D by manipulating the matrix properties. In the 3D microenvironment, macrophage-like RAW264.7 cells proliferated and formed spherical clusters by degrading the surrounding hydrogel network. Interestingly, we observed the significant upregulation of matrix metalloproteinases (MMPs) (i.e., MMP9 and MMP14) as well as M1 polarization markers (i.e., iNOS, COX2, TNF-α) in 3D, whereas M2 polarization markers (i.e., CD206, Arg1, TGF-β) were downregulated. Specifically, the expressions of both M1 and M2 markers were simultaneously increased in a stiff matrix compared to those of a soft matrix. In addition, matrix degradability significantly influenced the TNF-α secretion of encapsulated RAW264.7 cells. The MMP sensitivity of the hydrogel decreased TNF-α expression in a soft matrix, whereas it upregulated TNF-α in a stiff matrix compared to those of MMP-insensitive hydrogel. These findings suggest that the highly tunable poly(ethylene glycol) hydrogels can dictate macrophage behavior by altering the surrounding 3D microenvironment.

Soursop (Annona Muricata, Linn) Leaf Ethanol Extract Cream Application Affected the Expression of TNF-α and VEGF on Balb/C Mice Skin Exposed to Acute UVB

Background: Wound healing problems are often associated with TNF-α and VEGF expressions. However, the benefits and effects of ethanol extract cream from soursop leaves towards TNF-α and VEGF expression for burns healing process are yet to be known. Objective: To evaluate the benefits and influence of ethanol extract cream from soursop leaves in decreasing TNF-α expression and stimulate VEGF expression in BALB/c mice exposed to acute UVB radiation. Method: This study is a laboratory-based experimental study with post test only control group design. The subjects were 48 BALB/c female mice, divided into 24 subjects for TNF-α and 24 for the VEGF group, and were randomly distributed into 4 groups in each study, with each group containing 6 BALB/c mice. Group K was smeared with a base cream of 0.3 grams three times daily, while P1, P2 and P3 groups were each applied 0.3 grams of ethanol extract with consecutive consentrations of 2.5%, 5%, and 10% three times daily. On the sixth day, termination was performed on 24 mice to assess Tnf-α expression and on the eighth day also termination on 24 mice to assess VEGF expression. Preparation of substances was done using IHC method, analyzed using Kruskal Wallis test followed by Mann Whitney test.Results: The results of TNF-a and VEGF expression analysis based on Kruskal Wallis test obtained a p value of 0.0001 (p&lt;0.05) showing that the mean expression of TNF-a and VEGF among the four groups ware significantly different. Mann Whitney test results revealed that the differences in expression of TNF-a and VEGF between the two groups were all significant differences (p&lt;0.05). The mean TNF-a expression in the three treatment groups (PI, PII, PIII) ware significantly lower than the control group. The mean TNF-a expression in the PII and PIII treatment groups ware significantly lower than in the PI group, and the mean TNF-a expression in the PIII treatment group was significantly lower than the PII group. The mean VEGF expression in the three treatment groups (PI, PII, PIII) ware significantly higher than the control group. The mean VEGF expression in the PII and PIII treatment groups ware significantly higher than the PI group, and the mean VEGF expression in the PIII treatment group was significantly higher than in the PII group.Conclusion: Ethanol creams made of soursop leaves extract is proven to be useful and decreasing TNF-α expression and increase VEGF expression in BALB/c mice exposed to acute UVB rays.Keywords: Ethanol cream of soursop leaves extract, TNF-α expression, VEGF expression

The regulation mechanism of silencing the DUSP1 gene on the release of proinflammatory cytokines in mice with acute pancreatitis

Objective To investigate the regulation mechanism of silencing the DUSP1 gene on the release of proinflammatory cytokines in mice with acute pancreatitis(AP). Methods Two DUSP1-siRNA and one scramble siRNA sequences were designed, and the sequence with higher silence efficiency was selected.Mice models with AP were established, and KM mice were divided into 6 groups: control group, AP group, AP+ PD98059 group, AP+ scramble group, AP+ siRNA group and AP+ PD98059+ siRNA group.Expressions of proinflammatory cytokines tumor necrosis factor-α(TNF-α), interleukin(IL)-1β and IL-6 in serum were detected by using enzyme linked immunosorbent assay(ELISA) after 12 h, 24 h, 48 h of modeling.Serum amylase levels were detected.The mRNA expression levels of DUSP1, TNF-α, IL-1β and IL-6 in pancreatic tissues were detected by using quantitative real time polymerase chain reaction (qPCR). The protein expression levels of DUSP1, extracellular regulated protein kinases(ERK), c-Jun N-terminal kinase(JNK), p38, p-ERK, p-JNK and p-p38 in pancreatic tissues were detected by using Western blot. Results Compared with the control group, the other 5 groups displayed the increased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of DUSP1, TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there was a statistical significance (all P<0.05). Compared with the AP group, the AP+ PD98059+ siRNA group showed the decreased DUSP1 expression in tissues, and there was a statistical significance (all P<0.05); the AP+ PD98059 group had decreased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there were statistical significances (all P<0.05); while the opposite results were observed in the AP+ siRNA group with DUSP1 expression decreased. Conclusions The results support that silencing the DUSP1 gene promotes the release of proinflammatory cytokines through activating the mitogen-activated protein kinase signaling pathway in mice with AP. Key words: Mitogen-activated protein kinase signaling pathway; Acute pancreatitis; Proinflammatory cytokines; Lentiviral vectors; DUSP1 gene

Perinatal short-chain fructooligosaccharides program intestinal microbiota and improve enteroinsular axis function and inflammatory status in high-fat diet-fed adult pigs.

Perinatal nutrition programs physiologic and metabolic functions, with consequences on the susceptibility to develop metabolic diseases in adulthood. The microbiota represents a key factor of such programming. We investigated whether perinatal prebiotic [short-chain fructooligosaccharides (scFOS)] supplementation improved adult metabolic health in association with microbiota changes in pigs used as human model. Sows were supplemented with scFOS or not during the end of gestation and the entire lactation, and offspring received scFOS accordingly during 1 mo after weaning. Pigs were then fed a standard diet for 5 mo, followed by a high-fat diet for 3 mo once adults. Perinatal scFOS supplementation induced a persistent modulation of the composition of the fecal microbiota in adulthood, notably by increasing the Prevotella genus. Meanwhile, scFOS animals displayed improved capacity to secrete glucagon-like peptide-1 and improved pancreas sensitivity to glucose without any changes in peripheral insulin sensitivity. Perinatal scFOS supplementation also increased ileal secretory IgA secretion and alkaline phosphatase activity and decreased TNF-α expression in adipose tissue. In conclusion, perinatal scFOS supplementation induced long-lasting modulation of intestinal microbiota and had beneficial consequences on the host physiology in adulthood. Our results highlight the key role of perinatal nutrition on later microbiota and host metabolic adaptation to an unbalanced diet.-Le Bourgot, C., Ferret-Bernard, S., Apper, E., Taminiau, B., Cahu, A., Le Normand, L., Respondek, F., Le Huërou-Luron, I., Blat, S. Perinatal short-chain fructooligosaccharides program intestinal microbiota and improve enteroinsular axis function and inflammatory status in high-fat diet-fed adult pigs.

Overexpression of Sirtuin2 prevents high glucose-induced vascular endothelial cell injury by regulating the p53 and NF-κB signaling pathways.

To investigate the potential role and underlying mechanism of Sirtuin2 (SIRT2) in regulating high glucose (HG)-induced vascular endothelial cell injury by using human umbilical vein endothelial cells (HUVECs). SIRT2 mRNA and protein expression levels were decreased in HG-treated HUVECs. SIRT2 overexpression increased viability, decreased apoptosis and reduced levels of reactive oxygen species in HG-treated HUVECs. SIRT2 overexpression decreased TNF-α expression (146.5±22.8pg TNF-α ml-1) relative to that in the empty vector group (263.5±18.5pg TNF-α ml-1) and decreased MCP-1 expression (63.8±9.85pg MCP-1ml-1) relative to that in the empty vector group (105.8±8.5pg MCP-1ml-1). SIRT2 overexpression decreased the acetylation of p53 by 33% and decreased the acetylation of NF-κB p65 by 58% in HG-treated HUVECs. SIRT2 prevents HG-induced vascular endothelial cell injury through suppressing the p53 and NF-κB signaling pathways.