IL-10 Gene Expression Research Articles

Autophagy induction protects the perivascular adipose tissue during hyperaldosteronism-induced hypertension

Hyperaldosteronism (HA), characterized by an autonomous production of aldosterone, promotes increased blood pressure and cardiovascular damage. The perivascular adipose tissue (PVAT) participates of the control of vascular function and is inflamed in HA. Autophagy is a recycling process of unnecessary or dysfunctional components and it is important for inflammatory tolerance, but whether the autophagic flux is dysregulated in the PVAT during HA, which would contribute to the inflammation and dysfunction of this tissue, is still to be elucidated. Herein, we hypothesize that treatment with autophagy inducer will rescue the autophagic unbalance, subsequently decreasing inflammation, and improving PVAT function during HA. 10-12-weeks-old male C57BL/6J mice were infused with aldosterone for 14 days (600ug/Kg/day via osmotic mini-pump). Vascular function was studied in PVAT-intact thoracic aortae and blood pressure was analyzed by the tail-cuff. Brown adipocyte cells (WT1) were used to evaluate the molecular mechanisms by which HA induces PVAT dysfunction. HA promoted an increase in systolic blood pressure [mmHg – before HA: 123.8±0.9 vs. after HA: 149.5±1.1]. Furthermore, PVAT decreased the contractile response [Emax (% KCl)] in control arteries [Control PVAT (-): 137.2±1.9 vs. Control PVAT (+): 107.4±2.7], whereas HA promoted loss of PVAT anti-contractile effect [HA PVAT (-): 159.9±2.9 vs. HA PVAT (+): 164.1±3.4]. We found that PVAT from HA mice presents an unbalance in LC3II/LC3I ratio, an important autophagy marker, inflammation, characterized by elevated TNF-α, IL-6, IL-1β, IL-17, and IFN-γ gene expression, and elevated reactive oxygen species (ROS). To induce autophagy, we treated mice with TB-peptide (16mg/Kg/day via ip for 7 days). TB-peptide partially improved the PVAT function of mice with HA [HA PVAT (+): 122.5±4.1]. On mechanistic levels, aldosterone (100nM, 12-96 h) induced an oscillatory autophagy flux in WT1 cells, characterized by an initial increase in LC3II/LC3I ratio at 12 and 24 h, followed by a decrease at 48 and 96 h. These effects were not observed in the presence of the mineralocorticoid receptor antagonist (Eplerenone, 1μM). Aldosterone also led to the expression of pro-inflammatory genes (TNF-α, IL-6, IL-1β, IL-17, and IFN-γ) and elevated ROS, such effects were prevented by inducing autophagy with TB-peptide. In summary, these findings reveal a new mechanism in vascular injury associated with HA, aldosterone impairs autophagic flux in PVAT, inducing inflammation, oxidative stress, and dysfunction. Our study places PVAT and autophagic process as attractive targets to reduce cardiovascular risk in HA. FAPESP (2022/06639-2), NHLBI-R00 (R00HL14013903), AHA-CDA (CDA857268), Vascular Medicine Institute, the Hemophilia Center of Western Pennsylvania Vitalant, and Children's Hospital of Pittsburgh of the UPMC Health System. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Regulating the Regulators: determinants of lineage- and tissue-specific Il10expression in CD4 +T cells

Abstract IL-10 is an essential anti-inflammatory cytokine involved in mucosal immune regulation. Whereas many lineages of innate and adaptive immune cells can produce IL-10, a comprehensive understanding of the common, lineage-specific and tissue-specific regulatory mechanisms underlying Il10transcriptional regulation has remained elusive. Our lab recently identified the constitutive androstane receptor (CAR; encoded by Nr1i3)—a xenobiotic-sensing nuclear receptor (NR) with no previously known function in immune cells—as a selective regulator of Il10 gene expression in small intestinal (SI) type 1 regulatory (Tr1) cells. CAR induction of IL-10 expression in SI Tr1 cells stands in direct contrast with orthogonal mechanisms that underlie Il10gene regulation in large intestinal (LI) Foxp3+ T regulatory (Treg) cells, and that involve other nuclear receptors, such as vitamin D receptor (VDR). By using a combination of next-generation sequencing techniques (ATAC-seq, CITE-seq, ChIP-seq), I am assessing how, where and in which cell types CAR most prominently regulates Il10expression in vitroand in vivo. Our results are generating high resolution maps of Il10transcriptional regulatory networks that are differentially called upon in different cell types and discrete mucosal tissues to achieve the complex and dynamic regulation of Il10expression that is observed in vivo. Supported by grants from NIH (RO1 AI143821-03)

A Study on IFN-γ and IL-10 Gene Expression Changes in Gallus Gallus Domesticus Embryo Infected with Klebsiella Pneumoniae and its Possible Alteration by Bakreshwar Hot Spring Water

Background: Hot spring's water is enriched with minerals which has many therapeutic values. Bakreshwar is well known for its ten hot springs of varying temperatures (35°C-71°C). Bathing repeatedly in hot springs can help us to fight against skin infection, rheumatic disorders, joint pain etc. Objectives: The present study aimed to explore the changes in the gene expression of IL-10 and IFN-γ in the Gallus gallus domesticus embryo, infected with multi-drug-resistant Klebsiella pneumoniae and by using the Bakreshwar hot spring water to make any curative changes in the target gene expression or not. Methodology: 14 days old embryonated eggs of Gallus gallus domesticus was inoculated with freshly prepared 0.5 Mc Farland standard suspension of Klebsiella pneumoniae and curative set was treated with Bakreshwar Hot Spring water, collected from Agnikund (65°C). Then the embryonated eggs were harvested following the Institutional Ethical Committee guidelines and the allantoic fluid was collected. On the next day, RNA extraction was performed and cDNA synthesis followed by RT PCR was performed to study the cytokine gene expression. Results: After infection of chick embryo Gallus gallus domesticus with multi drug resistant Klebsiella pneumoniae, both IL-10 and IFN-γ gene expression was increased, but in curative sets that were challenged with Bakreshwar water, IL-10 gene expression was decreased and IFN-γ was increased. Conclusion: Hot spring water of Bakreshwar has antimicrobial effect on multi drug resistant Klebsiella pneumonia and it can normalize the cytokine balance with prolonged protection with raised IFN-γ.

Soluble adenylyl cyclase contributes to imiquimod-mediated inflammation and is a potential therapeutic target in psoriasis.

Cyclic AMP (cAMP) has a key role in psoriasis pathogenesis, as indicated by the therapeutic efficacy of phosphodiesterase inhibitors that prevent the degradation of cAMP. However, whether soluble adenylate cyclase (sAC) (encoded by the ADCY10 gene), which is an important source for cAMP, is involved in Th17 cell-mediated inflammation or could be an alternative therapeutic target in psoriasis is unknown. We have utilized the imiquimod model of murine psoriasiform dermatitis to address this question. Adcy10-/- mice had reduced erythema, scaling and swelling in the skin and reduced CD4+ IL17+ cell numbers in the draining lymph nodes, compared with wild-type mice after induction of psoriasiform dermatitis with imiquimod. Keratinocyte-specific knock out of Adcy10 had no effect on imiquimod-induced ear swelling suggesting keratinocyte sAC has no role in imiquimod-induced inflammation. During Th17 polarization in vitro, naive T cells from Adcy10-/- mice exhibited reduced IL17 secretion and IL-17+ T-cell proliferation suggesting that differentiation into Th17 cells is suppressed without sAC activity. Interestingly, loss of sAC did not impact the expression of Th17 lineage-defining transcription factors (such as Rorc and cMaf) but rather was required for CREB-dependent gene expression, which is known to support Th17 cell gene expression. Finally, topical application of small molecule sAC inhibitors (sACi) reduced imiquimod-induced psoriasiform dermatitis and Il17 gene expression in the skin. Collectively, these findings demonstrate that sAC is important for psoriasiform dermatitis in mouse skin. sACi may provide an alternative class of topical therapeutics for Th17-mediated skin diseases.

Short-term exposure to fumonisins and deoxynivalenol, on broiler growth performance and cecal Salmonella load during experimental Salmonella Enteritidis infection

Fumonisins (FUM) and deoxynivalenol (DON) are two common mycotoxins in poultry feed. Salmonella enterica ser. Enteritidis (S. Enteritidis) is a primary foodborne bacterium in broilers. This trial was conducted to evaluate the effects of naturally occurring FUM and DON and their combination at subclinical doses on broiler performance during a S. Enteritidis challenge. The experiment consisted of five treatments: NCC, no-challenge no-mycotoxin treatment; CC, Salmonella challenge+no-mycotoxin treatment; DON, DON 0.6 mg/kg+Salmonella challenge; FUM, FUM 14 mg/kg+Salmonella challenge; DON + FUM + T-2 + neosolaniol, DON 0.6 mg/kg+FUM 14 mg/kg+T-2 toxin 0.6 mg/kg+0.8 mg/kg neosolaniol+Salmonella challenge. On d 4, birds were challenged with either 0 or 1×109 CFU/mL S. Enteritidis orally. There were no significant effects on growth performance among treatments at 0, 3, 7, and 14 d of post-inoculation (dpi). On 14 dpi, the combined DON + FUM + T-2 + neosolaniol significantly increased the Salmonella load by 1.5 logs compared to the control groups (P < 0.05). FUM significantly increased the cecal tonsil IL-10 gene expression by 1.2-fold at 7 dpi (P < 0.05) and downregulated TNF-α by 1.8-fold on 14 dpi compared to the control, nonchallenge groups (P < 0.05). On 7 dpi, the combined DON + FUM + T-2 + neosolaniol reduced occludin by 4.4-fold (P < 0.05) when compared to the control groups. Similarly, combined DON + FUM+ T-2 + neosolaniol decreased zona-occluden transcription by 2.3 and 7.6-fold on 3 and 14 dpi, respectively (P < 0.05). Furthermore, combined DON + FUM + T-2 + neosolaniol decreased Claudin-1 by 2.2-fold and Claudin-4 by 5.1-fold on 14 dpi when compared to the control groups (P < 0.05). In conclusion, short-term exposure to a subclinical dose of combined DON + FUM + T-2 + neosolaniol had an impact on broiler intestinal tight junction proteins and cecal Salmonella abundance under experimental Salmonella challenge.

Immune Response Evaluation in Commercial Turkeys Affected with Clostridial Dermatitis.

Clostridial dermatitis (CD), caused by Clostridium septicum and Clostridium perfringens, is an economically important emerging disease of turkeys characterized by sudden deaths and necrotic dermatitis. Immune responses in CD-affected commercial turkeys are poorly understood. In the present study, C. septicum was isolated from CD-affected commercial turkeys during a recent outbreak, and the tissues (skin, muscle, and spleen) were collected and analyzed for immune gene expression, along with samples from clinically healthy birds. The results showed that CD-affected turkeys had significantly higher levels of IL-1β, IL-6, IFNγ, and iNOS transcripts in the skin, muscle, and spleen tissues compared to healthy birds. Affected turkeys also had a significantly elevated transcription of toll-like receptor (TLR21) gene in the skin and spleen tissues, suggesting a role for this receptor in the immune recognition. The expression of IL-4 and IL-13 genes in the spleen and muscle was also significantly higher in the affected birds. Additional birds from the same affected and healthy farms examined for serology revealed that the CD-affected turkeys had significantly higher levels of serum IgM and IgY antibodies. Furthermore, in vitro stimulation of MQ-NCSU macrophages with C. septicum led to a significant transcriptional upregulation of IL-1β and IFNγ genes, while the IL-10 gene expression was downregulated. The surface expression of MHC-II protein and cellular production of nitric oxide were also significantly increased in the C. septicum-stimulated macrophages, indicating cellular activation. Collectively, our findings suggest that the host responses in CD-affected turkeys involve a robust inflammatory response as well as a response mediated by IL4/IL-13 cytokines that may aid in antibody-mediated immunity.

Tyndallized Bacteria Preferentially Induce Human Macrophage M1 Polarization: An Effect Useful to Balance Allergic Immune Responses and to Control Infections

Macrophage polarization is a dynamic process through which macrophages acquire specific features whose extremes are represented by M1 and M2 polarization. Interleukin (IL)-6, IL-1β, IL-12 and IL-8 belong to M1 macrophages while transforming growth factor-beta (TGF-β belongs to M2 cytokines. M2 polarization prevalence is observed in allergic diseases. Tyndallization is a thermal process able to inactivate microorganisms and to allow their use for chronic respiratory disease treatment via immune response modulation. The present study explores the effects of a blend of tyndallized bacteria (TB) on macrophage polarization. THP-1-derived macrophages were exposed to different concentrations of TB (106, 5 × 106, 107, 5 × 107, 108 CFU/mL) and then cell viability and TB phagocytosis, and IL-8, IL-1β, IL-6, IL-12 and TGF-β1 gene expression and release were assessed. TB were tolerated, phagocyted and able to increase IL-8, IL-1β and IL-6 gene expression and release IL-12 gene expression, as well as decrease TGF-β1 gene expression and release. The effects on IL-8, IL-6 and TGF-β1 release were confirmed in human monocyte-derived macrophages (hMDMs) exposed to TB. In conclusion, TB promote M1 polarization, and this mechanism might have valuable potential in controlling allergic diseases and infections, possibly preventing disease exacerbations.

Expression of CD47 and its ligands in pregnant mice infected with Toxoplasma gondii during pregnancy

Expression of CD47 and its ligands in pregnant mice infected with Toxoplasma gondii during pregnancy

The anti-inflammatory effect of lutein in broilers is mediated by regulating Toll-like receptor 4/myeloid-differentiation-factor 88 signaling pathway

The anti-inflammatory role of lutein has been widely recognized, however, the underlying mechanism is still not fully elucidated. Hence, the effects of lutein on the intestinal health and growth performance of broilers and the action of mechanism were investigated. 288 male yellow-feathered broilers (1-day old) were randomly allocated to 3 treatment groups with 8 replicates of 12 birds each, and the control group was fed a broken rice-soybean basal diet, while the test groups were fed a basal diet added with 20 mg/kg and 40 mg/kg of lutein (LU20, LU40), respectively. The feeding trial lasted for 21 d. The results showed that 40 mg/kg lutein supplementation tended to increase ADFI (P=0.10) and ADG (P=0.08) of broilers. Moreover, the addition of lutein caused a decreasing trend of gene expression and concentration of proinflammatory cytokines IL-1β (P=0.08, P=0.10, respectively) and IL-6 (P=0.06, P=0.06, respectively) and also tended to decrease the gene expression of TLR4 (P=0.09) and MyD88 (P=0.07) while increasing gene expression and concentration of anti-inflammatory cytokines IL-4 and IL-10 (P < 0.05) in the jejunum mucosa of broilers. Additionally, lutein supplementation increased the jejunal villi height of broilers (P < 0.05) and reduced villi damage. The experiment in vitro showed that lutein treatment reduced the gene expression of IL-1β, IL-6, and IFN-γ in chicken intestinal epithelial cells (P < 0.05). However, this effect was diminished after knock-down of TLR4 or MyD88 genes using RNAi technology. In conclusion, lutein can inhibit the expression and secretion of proinflammatory cytokines in the jejunum mucosa and promote intestinal development of broilers, and the anti-inflammatory effect may be achieved by regulating TLR4/MyD88 signaling pathway.

Invited Perspective: Environmental Chemical-Sensing AHR Remains an Enigmatic Key Player in Toxicology.

Invited Perspective: Environmental Chemical-Sensing AHR Remains an Enigmatic Key Player in Toxicology.

Effect of transduced mesenchymal stem cells with IL-10 gene on control of allergic asthma.

Asthma is an important pulmonary disease associated with T helper lymphocyte (Th)2 dominant immune response, which can initiate allergic and inflammatory reactions. Interleukin (IL)-10 is the main immune suppressor cytokine, and mesenchymal stem cells (MSCs) have an immune-modulatory potential that can be transduced with the expression of the IL-10 gene to control pathophysiology of allergic asthma. Bone marrow's MSCs were isolated and transduced with the expression vector that contains the expressible IL-10 gene. Then, allergic asthma mouse model was produced and treated with manipulated MSCs. Methacholine challenge test; measurement of IL-4, IL-5, IL-8, IL-13, IL-25, and IL-33; and total and ovalbumin (OVA)-specific immunoglobulin (Ig)E levels were done. Hyperplasia of the goblet cell, secretion of mucus, and peribronchiolar and perivascular eosinophilic inflammation were evaluated in lung pathological sections. IL-25, IL-33, and total IgE levels; AHR; eosinophilic inflammation; hyperplasia of the goblet cell; and secretion of mucus could be controlled in M, MV, and MV-10 groups, and the control in the MV-10 group was strong compared to M and MV groups. MSCs have immune-modulatory capacity that can control allergic asthma pathophysiology, and this effect can be strengthened and reinforced by the expression of IL-10 gene.

The effect of different C. difficile MLST strains on viability and activity of macrophages

ObjectivesClostridioides difficile is the most common infectious agent of nosocomial diarrhea. C. difficile infection (CDI) pathogenesis and disease severity depend on its toxins (toxins A, B and binary) and on the host’s immune response, especially the innate immune system. The current study examined the efficacy of macrophage activity, macrophages viability and cytokine secretion levelsin response to different sequence type (ST) strains of C. difficile. MethodsRAW 264.7 macrophages were exposed to six different strains of C. difficile as well as to both toxins A and B and macrophage viability was measured. The levels of four secreted cytokines were determined by RT-PCR and ELISA. Morphological changes to the macrophages were investigated by fluorescent microscopy. ResultsStrains ST37 and ST42 affected macrophages’ vitality the most. Toxins A and B led to a significant reduction in macrophages' vitality at most time points. In addition, starting at 30-min post-exposure to 5 ng/μl of both toxins led to significant differences in macrophage viability versus at lower concentrations. Furthermore, cytokine secretion levels, including IL-12, IL-6 and TNF-α, increased dramatically when macrophages were exposed to strains ST42 or ST104. Finally, gene expression surveys point to increases in IL-12 gene expression in response to both ST42 and ST104. ConclusionsC. difficile strains with higher toxins levels induced an increased activation of the innate immune system and may activate macrophages more profoundly resulting in secretion of higher levels of pro-inflammatory cytokines. However, higher toxin levels may also damage macrophages’ normal skeletal structure, reducing macrophage viability.

Immunological and Molecular Evaluation of IL-5 in Asthma Patients

Asthma cause airway obstruction and are associated with chronic airway inflammation. Interleukin -5 is mainly produced by Th2 cells, innate lymphoid cells group 2 (ILC2), mast cells, natural killer T cells (NKT), and eosinophils themselves. Material and Method: A case-control study and the implementation steps are divided into 3 main parts, include: Patient selection and sampling, examining the intensity of gene expression. Result: The present study show that the expression of IL-5 gene has increased significantly in asthma. Regarding the intensity of IL-5 gene expression in the patient and control groups, a significant difference can be observed between the two groups in terms of gene expression intensity (PValue=0.0001). Conclusion: The results of the present study show that IL-5 gene expression has been significantly changed in asthma. In terms of the intensity of IL-5 gene expression in the patient and control groups, a significant difference is observed between the two groups (P-Value=0.0001), also the experiment and analysis of the obtained ROC curve shows that a significant change in IL-5 in Asthma.

Microbe-Derived Antioxidants Alleviate Liver and Adipose Tissue Lipid Disorders and Metabolic Inflammation Induced by High Fat Diet in Mice.

Obesity induces lipodystrophy and metabolic inflammation. Microbe-derived antioxidants (MA) are novel small-molecule nutrients obtained from microbial fermentation, and have anti-oxidation, lipid-lowering and anti-inflammatory effects. Whether MA can regulate obesity-induced lipodystrophy and metabolic inflammation has not yet been investigated. The aim of this study was to investigate the effects of MA on oxidative stress, lipid disorders, and metabolic inflammation in liver and epididymal adipose tissues (EAT) of mice fed with a high-fat diet (HFD). Results showed that MA was able to reverse the HFD-induced increase in body weight, body fat rate and Lee's index in mice; reduce the fat content in serum, liver and EAT; and regulate the INS, LEP and resistin adipokines as well as free fatty acids to their normal levels. MA also reduced de novo synthesis of fat in the liver and EAT and promoted gene expression for lipolysis, fatty acid transport and β-oxidation. MA decreased TNF-α and MCP1 content in serum, elevated SOD activity in liver and EAT, induced macrophage polarization toward the M2 type, inhibited the NLRP3 pathway, increased gene expression of the anti-inflammatory factors IL-4 and IL-13 and suppressed gene expression of the pro-inflammatory factors IL-6, TNF-α and MCP1, thereby attenuating oxidative stress and inflammation induced by HFD. In conclusion, MA can effectively reduce HFD-induced weight gain and alleviate obesity-induced oxidative stress, lipid disorders and metabolic inflammation in the liver and EAT, indicating that MA shows great promise as a functional food.

Additive beneficial effects of aerobic training and royal jelly on hippocampal inflammation and function in experimental autoimmune encephalomyelitis rats.

Although the beneficial role of training and the use of some antioxidants in physiological and psychological disorders in autoimmune diseases has been reported, the simultaneous effect of aerobic training (AT) and royal jelly (RJ) with different doses is not well understood. The present study aimed to investigate the impact of AT and RJ on inflammatory factors in the hippocampus, as well as depression and anxiety in the experimental autoimmune encephalomyelitis (EAE). Sprague-Dawley rats with EAE were assigned to seven groups: (1) EAE without any other intervention (EAE); (2) sham, receiving normal saline (Sh); (3) 50mg/kg RJ (RJ50); (4) 100mg/kg RJ (RJ100); (5) AT; (6) AT+RJ50; and (7) AT+RJ100. In addition, a healthy control group was assessed. EAE significantly increased interleukin 17 (IL-17), transforming growth factor-β (TGF-β) gene expression and immobilization time as well as anxiety and depression indices, and significantly decreased interleukin 10 (IL-10), compared to the control group. AT decreased significantly IL-17, TGF-β gene expression and immobilization time as well as anxiety and depression indices, while it significantly increased IL-10, compared to the EAE group. RJ50 and RJ100 decreased significantly IL-17, IL-23 gene expression, anxiety and depression indices, and significantly increased IL-10 compared to the EAE group. AT+RJ50 and AT+RJ100 significantly decreased IL-17, IL-23, and TGF-β and as well as anxiety and depression indices while significantly increasing IL-10 compared to the EAE group. The effects of AT+RJ100 on significant decreasing IL-17, IL-23, anxiety and depression and increasing TGF-β, IL-10 were more favorable than RJ50. AT and RJ improved inflammatory and regulatory factors of autoimmunity and reduced anxiety and depression. The RJ combined with AT induced additive effects while using RJ100 was more favorable than RJ50.

The Immunomodulatory Aspect of Quercetin Penta Acetate on Th17 Cells Proliferation and Gene Expression in Multiple Sclerosis.

The function of Th17 cells in the neuroinflammatory process in multiple sclerosis (MS) has been previously clarified. It has been suggested that Quercetin can influence MS due to a variety of anti-inflammatory effects. The present study aimed to examine in vitro immunomodulatory aspects of Quercetin Penta Acetate as a modified compound on Th17 cells of MS patients and also to compare its effects with Quercetin. In this experimental study, peripheral blood mononuclear cell (PBMCs) were isolated and stained with CFSE then, half-maximal inhibitory concentration (IC50) values were determined using different doses and times for Quercetin Penta Acetate, and Methyl Prednisolone Acetate. Th17 cell proliferation was analyzed by flow cytometry and the expression levels of IL-17 and RORc genes were assessed by real-time polymerase chain reaction (PCR) method. The results showed that IL-17 gene expression was inhibited by Quercetin Penta Acetate (P=0.0081), but Quercetin Penta Acetate did not have a significant inhibitory effect on Th17 cells proliferation (P= 0.59) and RORc gene expression (P=0.1), compared to Quercetin. Taken together, our results showed some immunomodulatory aspects of Quercetin Penta Acetate on Th17 cells are more effective than Quercetin and it could be considered in the treatment of MS.

Quercetin in combination with hyperbaric oxygen therapy synergistically attenuates damage progression in traumatic spinal cord injury in a rat model

BackgroundOxidative stress, inflammation and cell apoptosis are the most important destructive factors in the spread of damage following trauma to the spinal cord. Therefore, presently, we investigated the synergistic effects of quercetin along with hyperbaric oxygen therapy (HBOT) as strong antioxidant, anti-inflammatory and anti-apoptotic compounds in the recovery of traumatic spinal cord injury (TSCI) in a rat model. Material and methodsSeventy-five male mature Sprague-Dawley rats allocated into 5 groups, including: Sham group (SG), TSCI group, Quercetin group (underwent TSCI and received quercetin), HBOT group (underwent TSCI and received HBOT), and Quercetin+ HBOT group (underwent TSCI and received quercetin plus HBOT). Finally, the spinal cord samples at the traumatic site were harvested and various characteristics were evaluated, including the total volumes of the spinal cord and its central cavity as well as the numerical density of neuron and glial cells by stereological method, oxidant (malondialdehyde; MDA) and antioxidant (glutathione; GSH, superoxide dismutase; SOD and catalase; CAT) factors by biochemical method, molecular levels of IL-10, TNF-α and IL-1β by qRT-PCR method, and cell apoptosis by immunohistochemistry method against Caspase-3 antibody. Furthermore, Basso-Beattie-Bresnahan (BBB) and electromyography latency (EMG Latency) tests were performed to evaluate neurological functions. ResultsFindings demonstrated that the stereological characteristics, biochemical factors (except MDA), expression of IL-10 gene and behavioral functions were significantly better in Quercetin, HBOT and Quercetin+HBOT groups than TSCI group, and were greater in Quercetin+HBOT ones (P < 0.05). While MDA levels, expression of TNF-α and IL-1β genes as well as the density of apoptotic cells significantly more decreased in Quercetin+HBOT group compared to other treated groups (P < 0.05). ConclusionOverall, co-administration of quercetin with HBOT has synergistic neuroprotective effects in animals underwent TSCI.

Low levels of TNFA gene expression seem to favor the development of pulmonary tuberculosis in a population from the Brazilian Amazon

TNF-α is a Th1 cytokine profile active in the control of Mycobacterium tuberculosis infection, IL-10 is associated with persistence of bacterial infection. The aim of the study was to investigate the association of TNFA -308G/A and IL10 -819C/T polymorphisms and TNFA and IL10 gene expression levels with pulmonary and extrapulmonary tuberculosis (n = 200) and control (n = 200). The individuals were submitted to genotyping and quantification of gene expression performed by real-time quantitative polymerase chain reaction (qPCR). No association was observed between the frequencies of polymorphisms evaluated and pulmonary tuberculosis. The frequency of polymorphic genotypes for TNFA -308G/A were associated with the extrapulmonary tuberculosis (p = 0.0445). The levels of TNFA expression were lower in the pulmonary tuberculosis group than in the control (p = 0.0009). There was a positive correlation between the levels of TNFA and IL10 in patients with pulmonary tuberculosis (r = 0.560; p = 0.0103). Reduced levels of TNFA expression may promote the formation of an anti-inflammatory microenvironment, favoring the persistence of the bacillus in the host, contributing to the establishment of pulmonary tuberculosis.

Cytokine expression profiles in white blood cells of patients with small fiber neuropathy

BackgroundThe role of cytokines in the pathophysiology, diagnosis, and prognosis of small fiber neuropathy (SFN) is incompletely understood. We studied expression profiles of selected pro- and anti-inflammatory cytokines in RNA from white blood cells (WBC) of patients with a medical history and a clinical phenotype suggestive for SFN and compared data with healthy controls.MethodsWe prospectively recruited 52 patients and 21 age- and sex-matched healthy controls. Study participants were characterized in detail and underwent complete neurological examination. Venous blood was drawn for routine and extended laboratory tests, and for WBC isolation. Systemic RNA expression profiles of the pro-inflammatory cytokines interleukin (IL)-1ß, IL-2, IL-8, tumor necrosis factor-alpha (TNF) and the anti-inflammatory cytokines IL-4, IL-10, transforming growth factor beta-1 (TGF) were analyzed. Protein levels of IL-2, IL-8, and TNF were measured in serum of patients and controls. Receiver operating characteristic (ROC)-curve analysis was used to determine the accuracy of IL-2, IL-8, and TNF in differentiating patients and controls. To compare the potential discriminatory efficacy of single versus combined cytokines, equality of different AUCs was tested.ResultsWBC gene expression of IL-2, IL-8, and TNF was higher in patients compared to healthy controls (IL-2: p = 0.02; IL-8: p = 0.009; TNF: p = 0.03) and discriminated between the groups (area under the curve (AUC) ≥ 0.68 for each cytokine) with highest diagnostic accuracy reached by combining the three cytokines (AUC = 0.81, sensitivity = 70%, specificity = 86%). Subgroup analysis revealed the following differences: IL-8 and TNF gene expression levels were higher in female patients compared to female controls (IL-8: p = 0.01; TNF: p = 0.03). The combination of TNF with IL-2 and TNF with IL-2 and IL-8 discriminated best between the study groups. IL-2 was higher expressed in patients with moderate pain compared to those with severe pain (p = 0.02). Patients with acral pain showed higher IL-10 gene expression compared to patients with generalized pain (p = 0.004). We further found a negative correlation between the relative gene expression of IL-2 and current pain intensity (p = 0.02). Serum protein levels of IL-2, IL-8, and TNF did not differ between patients and controls.ConclusionsWe identified higher systemic gene expression of IL-2, IL-8, and TNF in SFN patients than in controls, which may be of potential relevance for diagnostics and patient stratification.

Evaluation of You-Gui-Wan critical compounds inhibiting ALOX-5 and HDC gene expression in RBL-2H3 cells using a fractional factorial design

Ethnopharmacological relevanceThe traditional Chinese medicine (TCM) You-Gui-Wan (YGW) has been used to treat asthma for hundreds of years. Aim of the studyYGW is composed of 10 types of medicinal materials. However, the immune mechanism of YGW in asthma treatment has not been elucidated. Therefore, this study investigated asthma symptoms attenuated by YGW and the underlying immune regulatory mechanism. Materials and methodsIntratracheal (i.t.) stimulation of BALB/c mice with Dermatophagoides pteronyssinus (Der p) was performed once per week (40 μL, 2.5 μg/μL). For six consecutive weeks, different doses of YGW (0.2 g/kg and 0.5 g/kg) were orally administered 30 min before stimulation with Der p. After the last stimulation, airway hyperreactivity, lung gene expression, and total immunoglobulin E (IgE) in blood were evaluated using a whole-body plethysmograph system, real-time PCR, and ELISA, respectively. In addition, DNP-IgE/DNP-BSA was added to stimulate mast cells (RBL-2H3), and YGW or various compound compositions (Trial) were added to RBL-2H3 cells for 30 min to evaluate the effects of the drug on mast cell degranulation and on gene expression. JMP 5.1 software was used to design and analyze YGW's critical compounds by which it inhibited ALOX-5 and HDC gene expression in RBL-2H3 cells. ResultsYGW significantly decreased serum total IgE levels and airway hyperresponsiveness in asthmatic mice. YGW also reduced the gene expression of IL-6, TNF-α, IL-4, IL-13, and COX-2 in the lungs of asthmatic mice and RBL-2H3 cells. YGW and the compound (Trial 21) present in YGW inhibited the gene expression of ALOX-5 and HDC in RBL-2H3 cells. ConclusionThe experimental results indicate that YGW exhibits anti-airway hyperresponsiveness and specific immunomodulatory effects. In addition, YGW synergistically inhibits ALOX-5 and HDC gene expression in mast cells through a combination of 21 compounds, including luteolin, quercetin, and β-carotene.