Nuclear NF-κB Research Articles

Circ-Sirt1 controls NF-κB activation via sequence-specific interaction and enhancement of SIRT1 expression by binding to miR-132/212 in vascular smooth muscle cells.

NF-κB-mediated inflammatory phenotypic switching of vascular smooth muscle cells (VSMCs) plays a central role in atherosclerosis and neointimal formation. However, little is known about the roles of circRNAs in the regulation of NF-κB signaling. Here, we identify the involvement of circ-Sirt1 that was one of transcripts of SIRT1 host gene in VSMC inflammatory response and neointimal hyperplasia. First, in the cytoplasm, circ-Sirt1 directly interacts with and sequesters NF-κB p65 from nuclear translocation induced by TNF-α in a sequence-dependent manner. The inhibitory complex of circ-Sirt1-NF-κB p65 is not dependent on IκBα. Second, circ-Sirt1 binds to miR-132/212 that interferes with SIRT1 mRNA, and facilitates the expression of host gene SIRT1. Increased SIRT1 results in deacetylation and inactivation of the nuclear NF-κB p65. These findings illustrate that circ-Sirt1 is a novel non-coding RNA regulator of VSMC phenotype.

The Cellular Uptake and Apoptotic Efficiency of Colchicine is Correlated with Downregulation of MMP-9 mRNA Expression in SW480 Colon Cancer Cells.

Colchicine, a tricyclic alkaloid, is commonly used in treatment due to its antiinflammatory and anti-fibrotic effects. Besides its toxicity at high doses, colchicine is reported for its potential anticancer effects at lower concentrations. The present study aimed to evaluate the anticancer effects of colchicine in SW480 cells. The effect of colchicine on cell proliferation was determined by MTT assay. The cellular colchicine uptake was measured by HPLC analysis. The apoptotic effects was evaluated by annexin v binding assay and MMP-9 mRNA expression was determined by RT-PCR experiments. Colchicine showed significant cytotoxicity at 10 ng/ml and higher concentrations and caused a cell cycle arrest of SW480 cells at G2/M phase. The results of HPLC analysis showed that colchicine uptake was increased in correlation with treated concentrations. Colchicine concentrations have increased the amount of apoptotic cell population. The elisa and western blot measurements showed that colchicine led to nuclear translocation of NF-κB proteins and increased caspase levels. The real time PCR experiments showed that colchicine has inhibitory effect on MMP-9 mRNA expression in a concentration dependent manner. These results illustrated that low dose colchicine efficiently induced cell death and apoptosis of SW480 cells and the inhibition of MMP-9 mRNA levels was significantly correlated with the amount of cellular colchicine uptake, suggesting that colchicine has a potential value in the treatment of human colorectal cancer.

Content of interleukins-4, 6 and nuclear transcription factor NF-κB in children with atopic bronchial asthma depending on Ile50Val polymorphism of IL4RA gene, severity of the disease course and level of its control

Many studies on polymorphic variants of cytokines genes indicate that their role in bronchial asthma development in children is still not fully understood. Aim. To determine the indexes of interleukins – 4, 6 and nuclear transcription factor (NF-κB) content in patients with atopic bronchial asthma (BA) depending on Ile50Val polymorphism of IL4RA gene, severity of the disease course and level of its control. Materials and methods. Alleles and genotypes were distributed according to the Ile50Val polymorphism of the IL4RA gene in 75 patients with atopic bronchial asthma and 25 healthy school-age children. Genotyping of the IL4RA gene was performed using a polymerase chain reaction (PCR). Biochemical studies on the interleukin-4 (IL-4), interleukin-6 (IL-6) levels and the content of nuclear factor-kappa B (NF-κB) determination were performed using ELISA kits. The results of analysis revealed an increase in NF-κB level in patients with intermittent BA carrying A/A, A/G and G/G genotypes relative to IL-4 content: in the A/A carriers by 1.65 times (P 0.05). Conclusions. Thus, in children with BA and Ile50Val polymorphism of IL4RA gene, the activity of NF-κB has a significant effect and its content exceeds the levels of IL-4 and IL-6 in the genotypes A/A and A/G (P 0.05).

Over expression of IL-32 in rheumatoid arthritis modulates NF-κB activity

IL-32 is pro-inflammatory cytokine characterized by the induction of nuclear factor NF-κB activation and an important role in key cellular processes related to inflammation. We investigated whether the IL-32 expression may be involved in the mehanistic of rheumatoid arthritis through modulates the activity of NF-κB. We report that the concentrations of IL-32 are higher in the serums of RA patients than of control subjects. Results show that IL-32 induces the production of the proinflammatory cytokines IL-1-beta, IL-6, and increased NF-κB activation. We concluded that these data show that IL-32 is constitutively produced by RA synoviocytes, induces proinflammatory cytokine secretion, and activation NF-κB. IL-32 thereby may constitute a promising target to treat RA.

Effects of Systematic Shortening of Noncovalent C8 Side Chain on the Cytotoxicity and NF-κB Inhibitory Capacity of Pyrrolobenzodiazepines (PBDs).

The systematic shortening of the noncovalent element of a C8-linked pyrrolobenzodiazepine (PBD) conjugate (13) led to the synthesis of a 19-member library of C8-PBD monomers. The critical elements of 13, which were required to render the molecule cytotoxic, were elucidated by an annexin V assay. The effects of shortening the noncovalent element of the molecule on transcription factor inhibitory capacity were also explored through an enzyme-linked immunosorbent assay-based measurement of nuclear NF-κB upon exposure of JJN-3 cells to the synthesized molecules. Although shortening the noncovalent interactive element of 13 had a less than expected effect upon compound cytotoxicity due to reduced DNA interaction, the transcription factor inhibitory capacity of the molecule was notably altered. This study suggests that a relatively short noncovalent side chain at the C8 position of PBD is sufficient to confer cytotoxicity. The shortened PBD monomers provide a new ADC payload scaffold because of their potent cytotoxicity and drug-like properties.

Exogenous ghrelin ameliorates acute lung injury by modulating the nuclear factor κB inhibitor kinase/nuclear factor κB inhibitor/nuclear factor κB pathway after hemorrhagic shock

Previous studies have shown that ghrelin, a peptide produced in the stomach, attenuates acute lung injury (ALI) in various animal models, and that some of these effects are associated with inhibition of the nuclear factor κB signaling pathway. This study investigated whether ghrelin exerts beneficial effects on hemorrhagic shock (HS)-induced ALI by modulating nuclear factor κB inhibitor kinase/nuclear factor κB inhibitor/nuclear factor κB (IKK/IκBα/NF-κB) pathway activity. HS was induced in male SD rats by withdrawing blood to a mean arterial pressure (MAP) of 40 mm Hg for 1 h; rats then received ghrelin (10 nmol/kg) or vehicle intravenously and were resuscitated with the shed blood and an equal volume of Ringer lactate solution followed by observation for 2 h. After resuscitation, samples were collected and analyzed for lung histopathology, wet to dry weight ratio (W/D), bronchoalveolar lavage fluid (BALF) protein, neutrophil infiltration, plasma inflammatory cytokines (TNF-α and IL-6), and cytoplasmic phosphorylated IKKβ, IκBα, phosphorylated IκBα and nuclear NF-κB expression. Compared to those in the two sham groups, lung injury, W/D, BALF protein, neutrophil infiltration, plasma TNF-α and IL-6 levels, and IKK/IκBα/NF-κB pathway activation were significantly increased in HS rats. After ghrelin administration, all parameters analyzed were decreased compared to those without ghrelin in HS rats. Moreover, ghrelin alleviated the decreased MAP after resuscitation compared to that in HS rats. Exogenous ghrelin attenuates the inflammatory response and acute lung injury after HS. These beneficial effects appear to be mediated through inhibition of IKK/IκBα/NF-κB signaling.

Complement regulatory protein CD59a plays a protective role in immune liver injury of trichloroethylene-sensitized BALB/c mice

Trichloroethylene (TCE) is a major occupational and environmental chemical compound which causes occupational dermatitis medicamentosa-like of TCE with severe liver damage. Our previous studies showed that complement activation was a newly recognized mechanism for TCE-induced liver damage. The objective of this study was to explore the role of the key complement regulatory protein, CD59a, in TCE-induced immune liver injury. We firstly evaluated the changes of CD59a expression in liver tissue and then investigated if the changes were associated with membrane attack complex (MAC) formation, nuclear factor kappa B (NF-κB) activation and liver damage in BALB/c mice model of TCE-induced skin sensitization in the absence or presence of soluble recombinant rat CD59-Cys. The results showed that low expression of CD59a accompanied by MAC deposition in the liver of TCE-sensitized BALB/c mice, which was consistent in time. In addition, activation of NF-κB pathway, upregulation of inflammatory cytokine and liver damage also occured. Additional experiment showed that recombinant rat sCD59-Cys alleviated inflammation and liver damage in TCE-sensitized BALB/c mice. Moreover, recombinant rat sCD59-Cys reduced MAC formation and inhibited NF-κB activation measured by P-IκBα and nuclear NF-κB p65 in the liver of TCE-sensitized BALB/c mice. In conclusion, recombinant rat sCD59-Cys plays a protective role in immune liver injury of TCE-sensitized BALB/c mice.

Role of NF-κB activation in mouse bone marrow stromal cells exposed to 900 MHz radiofrequency fields (RF)

ABSTRACTNuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a primary transcription factor which plays a key role in several cellular processes including proliferation and survival. It is well-known that exposure to non-ionizing radiofrequency fields (RF), which are ubiquitous, interact with cellular components. The aim of the study was thus to examine whether exposure of mouse bone marrow stromal cells (BMSC) to RF also resulted in cellular interactions. BMSC were exposed to 900 MHz RF at 120 μW/cm2 power intensity for 4 hr/day for 5 consecutive days. The relative protein expression levels of NF-κB in the cytoplasm and nucleus of RF-exposed cells were compared to non-RF-exposed controls. At 30 min post-RF exposure a significant decrease in protein expression of NF-κB in the cytoplasm was accompanied by a concomitant increase in nuclear NF-κB protein expression levels. Similar responses were noted in the cytoplasm and nuclear NF-κB levels at 2 hr with a return to control concentrations in primary transcription factor at 24 hr post-RF treatment. Daily incubation of BAY 11–7082 an inhibitor of NF-κB for 90 min for 5 days followed by RF each day prevented the fall in cytoplasmic NF-κB and rise in nuclear primary transcription factor at 30 min and 2 hr. There were no marked alterations at 24 hr. Data showed that the effects of RF treatment on BMSC involved transient activation of NF-κB which may be attributed to RF-mediated cellular perturbation as evidenced by consequences of BAY 11–7082 inhibition.

Treatment With a Flavonoid-Rich Fraction of Bergamot Juice Improved Lipopolysaccharide-Induced Periodontitis in Rats.

Objective: In this study, we investigated the effects of a flavonoid-rich fraction of Bergamot juice (BJe) in rats subjected to experimental periodontitis induced by a single intragingival injection of lipopolysaccharides (LPS).Main Methods: Periodontitis was induced by a single intragingival injection of 1 μl LPS (10 μg/μl) derived from Salmonella typhimurium in sterile saline solution. The injection was made in the mesolateral side at the interdental papilla between the first and the second molar. Fourteen days after LPS injection, we performed radiographic analyses and then we surgically removed the gingivomucosal tissue surrounding the mandibular first molar for histological, immunohistochemical and molecular analysis.Results: LPS significantly induced oedema, tissue damage and increased neutrophil infiltration. At molecular level, we found increased NF-κB translocation as well as raised both TNF-α and IL-1β expression, other than modulation of apoptosis-associated proteins. Moreover, the increased myeloperoxidase activity was associated with up-regulation of adhesion molecules. Immunohistochemical analysis for nitrotyrosine and poly ADP-ribose displayed an intense staining in the gingivomucosal tissue. Oral administration of BJe for 14 consecutive days reduced tissue injury and several markers of gingival inflammation including nuclear NF-κB translocation, cytokines expression, myeloperoxidase activity and the expression of some adhesion molecules such as ICAM and P-selectin. BJe also decreased both nitrosative stress and PARP positive staining. Moreover, it caused down-regulation of Bax and up-regulation of Bcl-2 expression.Conclusion: Our findings demonstrate that BJe improves LPS-induced periodontitis in rats by reducing the typical markers of inflammation, thus suggesting its potential in the treatment of periodontal diseases.

A Multicenter Prospective Study of First-Line Antibiotic Therapy for Early-Stage Gastric Mucosa-Associated Lymphoid Tissue (MALT) Lymphoma and Diffuse Large B-Cell Lymphoma with Histological Evidence of MALT

Background: To evaluate the efficacy of first-line Helicobacter pylori (HP) eradication (HPE) therapy against HP-positive gastric mucosa-associated lymphoid tissue (MALT) lymphoma and diffuse large B-cell lymphomas (DLBCL) with histological evidence of MALT (DLBCL[MALT]). Methods: Patients with gastric DLBCL(MALT) (n=10) and gastric MALT lymphoma (n=36) who had participated in a multicenter prospective study on first-line HPE therapy, conducted by the Taiwan Cooperative Oncology Group between December-2006 and September-2014, were enrolled. The HPE regimen consisted of omeprazole, amoxicillin, and clarithromycin for 14 days. The histological responses of tumors were re-assessed using the Groupe d'Etude des Lymphomes de l'Adult (GELA) scoring system. The primary end point was the therapeutic efficacy (complete remission, CR) of first-line HPE. The levels of expression of cytotoxin-associated gene A (CagA), BCL10, and NF-κB and the presence of t(11;18)(q21;q21) associated with HP-independence (lack of lymphoma regression after HPE) were assessed. Findings: Of the total 46 patients, 34 (73.9 %) achieved CR, including eight with DLBCL(MALT) and 26 with MALT lymphoma. The CR rates for patients with stages IE (n=40) and IIE1 (n=6) tumors were 75.0 % and 66.7 %, respectively. The frequency of CagA expression was significantly higher (85.3 % [29/34] vs 33.3 % [4/12]; p=0.001), whereas the frequency of nuclear BCL10 expression was significantly lower (8.8 % [3/34] vs 83.3 % [10/12]; p<0.0001), in HP-dependent tumors than in HP-independent tumors. The expression of nuclear NF-kB was significantly associated with HP-independence (p=0.0001). The API2-MALT1 fusion transcript was detected in 4 (33.0 %) of the 12 HP-independent MALT lymphomas, but in none of the 10 DLBCL(MALT) tumors. Interpretation: HPE is considered a first-line treatment for early-stage HP-positive DLBCL(MALT) as well as MALT lymphoma of the stomach. BCL10, and/or NF-kB are potential therapeutic targets in these lymphoma subtypes. Trial Registration Number: This trial is registered with ClinicalTrials.gov, number NCT00327132. Funding Statement: This study was sponsored, conducted and analyzed by the TCOG and following research grants: MOST 107-2314-B-002 -217-MY3 from Ministry of Science and Technology, Taiwan, and MOHW107-TDU-B-211-123002 from Ministry of Health and Welfare, Taiwan. Declaration of Interests: All authors declare they have no competing interests. Ethics Approval Statement: All experimental protocols were approved by the Institutional Review Board (IRB) of the Research Ethical Committee of National Taiwan University Hospital (NTUH) (NTUH IRB numbers: 950606).

Clofibrate Treatment Decreases Inflammation and Reverses Myocardial Infarction-Induced Remodelation in a Rodent Experimental Model.

Myocardial infarction (MI) initiates an inflammatory response that promotes both beneficial and deleterious effects. The early response helps the myocardium to remove damaged tissue; however, a prolonged later response brings cardiac remodeling characterized by functional, metabolic, and structural pathological changes. Current pharmacological treatments have failed to reverse ischemic-induced cardiac damage. Therefore, our aim was to study if clofibrate treatment was capable of decreasing inflammation and apoptosis, and reverse ventricular remodeling and MI-induced functional damage. Male Wistar rats were assigned to (1) Sham coronary artery ligation (Sham) or (2) Coronary artery ligation (MI). Seven days post-MI, animals were further divided to receive vehicle (V) or clofibrate (100 mg/kg, C) for 7 days. The expression of IL-6, TNF-α, and inflammatory related molecules ICAM-1, VCAM-1, MMP-2 and -9, nuclear NF-kB, and iNOS, were elevated in MI-V. These inflammatory biomarkers decreased in MI-C. Also, apoptotic proteins (Bax and pBad) were elevated in MI-V, while clofibrate augmented anti-apoptotic proteins (Bcl-2 and 14-3-3ε). Clofibrate also protected MI-induced changes in ultra-structure. The ex vivo evaluation of myocardial functioning showed that left ventricular pressure and mechanical work decreased in infarcted rats; clofibrate treatment raised those parameters to control values. Echocardiogram showed that clofibrate partially reduced LV dilation. In conclusion, clofibrate decreases cardiac remodeling, decreases inflammatory molecules, and partly preserves myocardial diameters.

Fold-Change Detection of NF-κB at Target Genes with Different Transcript Outputs

The transcription factor nuclear factor (NF)-κB promotes inflammatory and stress-responsive gene transcription across a range of cell types in response to the cytokine tumor necrosis factor (TNF). Although NF-κB signaling exhibits significant variability across single cells, some target genes supporting high levels of TNF-inducible transcription exhibit fold-change detection of NF-κB, which may buffer against stochastic variation in signaling molecules. It is unknown whether fold-change detection is maintained at NF-κB target genes with low levels of TNF-inducible transcription, for which stochastic promoter events may be more pronounced. Here, we used a microfluidic cell-trapping device to measure how TNF-induced activation of NF-κB controls transcription in single Jurkat T cells at the promoters of integrated HIV and the endogenous cytokine gene IL6, which produce only a few transcripts per cell. We tracked TNF-stimulated NF-κB RelA nuclear translocation by live-cell imaging and then quantified transcript number by RNA FISH in the same cell. We found that TNF-induced transcript abundance at 2 h for low- and high-abundance target genes correlates with similar strength with the fold change in nuclear NF-κB. A computational model of TNF-NF-κB signaling, which implements fold-change detection from competition for binding to κB motifs, could reproduce fold-change detection across the experimentally measured range of transcript outputs. However, multiple model parameters affecting transcription had to be simultaneously varied across promoters to maintain fold-change detection while also matching other trends in the single-cell data for low-abundance transcripts. Our results suggest that cells use multiple biological mechanisms to tune transcriptional output while maintaining robustness of NF-κB fold-change detection.

Phycocyanin attenuates X-ray-induced pulmonary inflammation via the TLR2-MyD88-NF-κB signaling pathway

Phycocyanin (PC), a natural algal protein, is reported for having anti-oxidant and antiinflammatory properties. We investigated its ability to attenuate lung inflammation in mice subjected to X-ray radiation. Male C57BL/6 mice were assigned to the control, total body irradiation, PC pretreatment, and PC treatment groups. Mice in the PC pretreatment group were gavaged with 200 mg/kg PC for 7 consecutive days before irradiation, and those in the PC treatment group were gavaged with 200 mg/kg PC for 7 consecutive days after irradiation. Lungs were collected on Day 7 after irradiation exposure. Hematoxylin and eosin staining of mouse lung sections showed considerable inflammation damage 7 days after irradiation compared with the control lung but a reduction in pathological injury in the PC treatment group. Pretreatment or treatment with PC significantly decreased levels of interleukin-6 and tumor necrosis factor-α in the lung, and also increased the relative mRNA expression of superoxide dismutase and glutathione. In vivo, PC significantly reduced the expression of Toll-like receptor TLR2, myeloid differentiation primary response Myd88, and nuclear factor NF-κB, at both the transcriptional and translation level. Taken together, these data indicated that PC attenuated lung inflammatory damage induced by radiation by blocking the TLR2-MyD88-NF-κB signaling pathway. Therefore, PC could be a protective agent against radiation-induced inflammatory damage in normal tissues.

Role of NF-κB activation in mouse bone marrow stromal cells exposed to 900-MHz radiofrequency fields (RF)

ABSTRACTNuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a primary transcription factor which plays a key role in several cellular processes including proliferation and survival. It is well known that exposure to non-ionizing radiofrequency fields (RFs), which are ubiquitous, resulted in interaction with cellular components. The aim of the study was thus to examine whether exposure of mouse bone marrow stromal cells (BMSC) to RF also resulted in cellular interactions. BMSC were exposed to 900 MHz RF at 120 μW/cm2 power intensity for 4 hr/day for 5 consecutive days. The relative protein expression levels of NF-κB in the cytoplasm and nucleus of RF-exposed cells were compared to non-RF-exposed controls. At 30-min post-RF exposure, a significant decrease in protein expression of NF-κB in the cytoplasm was accompanied by a concomitant increase in nuclear NF-κB protein expression levels. Similar responses were noted in the cytoplasm and nuclear NF-κB levels at 2 hr with a return to control concentrations in primary transcription factor at 24-hr post-RF treatment. Daily incubation of BAY 11-7082, an inhibitor of NF-κB, for 90 min for 5 days followed by RF each day prevented the fall in cytoplasmic NF-κB and rise in nuclear primary transcription factor at 30 min and 2 hr. There were no marked alterations at 24 hr. Data showed that the effects of RF treatment on BMSC involved transient activation of NF-κB which may be attributed to RF-mediated cellular perturbation as evidenced by consequences of BAY 11-7082 inhibition.

Anti-inflammatory effect of aged black garlic on 12-O-tetradecanoylphorbol-13-acetate-induced dermatitis in mice.

BACKGROUND/OBJECTIVESAlthough aged black garlic has various biological activities such as anti-allergy, anti-inflammation and neuroprotection, effect of aged black garlic on chemically contact dermatitis is unclarified.MATERIALS/METHODSTo evaluate anti-dermatitic activity of aged black garlic extract, we investigated effects of a fraction of aged black garlic extract (BG10) on both in vivo and in vitro.RESULTSBG10 almost inhibited formation of nitric monoxide and interleukin-6 (IL-6; IC50, 7.07 µg/mL) at 25 µg/mL, and dose-dependently reduced production of tumor necrosis factor-α (TNF-α; IC50, 52.07 µg/mL) and prostaglandin E2 (IC50, 38.46 µg/mL) in lipopolysaccharide-stimulated RAW264.7 cells. In addition, BG10 significantly inhibited the expression of inducible nitric oxide synthase, cyclooxygenase-2 and nuclear NF-κB, and improved that of cytosolic levels of NF-κB and IκBα in the cells. Consistent with in vitro studies, BG10 (0.5 mg/mL) not only reduced ear edema but also suppressed the formation of IL-6 and TNF-α induced by 12-O-tetradecanoylphorbol-13-acetate in ear tissues of mice.CONCLUSIONSThese findings suggest BG10 has anti-dermatitic activity through inhibiting activation of macrophages. Therefore, such effects of BG10 may provide information for the application of aged black garlic for prevention and therapy of contact dermatitis.

The Inhibitory Properties of Ethanol Extracts of Some Culinary-Medicinal Mushrooms on the Secretion of Interleukin-8 and Vascular Endothelial Growth Factor by PC3 Cancer Cells.

Interleukin (IL)-8, a cytokine produced by immune and non-immune cells, induces angiogenesis via increased vascular endothelial growth factor (VEGF) secretion; both cytokines promote tumor growth. IL-8 and VEGF plasma levels correlate with prostate cancer severity, suggesting that therapeutic options aimed at their downregulation may modulate tumor growth. Available data suggest that Agaricus bisporus (white button mushroom [WBM]) extracts inhibit cancer cell proliferation through aromatase inhibition. However, the extent to which they affect IL-8 and VEGF remains to be elucidated. The aims of this study were to (1) investigate the antiproliferative properties of WBM, brown A. bisporus (portabella), and Lentinus edodes (shiitake mushroom) on PC3 cancer cells; (2) demonstrate that these properties are exerted through the regulation of both IL-8 and VEGF; and (3) determine the role of NFκB activation in the antiproliferative process of mushroom extracts. Cytokine secretion in the supernatant, NFκB activity, and cell proliferation were measured in PC3 cells incubated with 0-100 µg/mL of ethanol extracts of mushrooms. Mushroom extracts decreased IL-8 secretion and cell proliferation (P < .05), and also tended to decrease VEGF (P < .09). Decreased cell proliferation did not appear to result from cell death because trypan blue exclusion tests showed comparable cell viability among cultures. Mushroom extracts also decreased nuclear and total NFκB activity, and the ratio of nuclear to cytoplasmic activity (P < .05) suggesting altered translocation from the cytoplasm to the nucleus. Our data suggest that the three types of studied mushrooms may modulate tumor growth through inhibition of IL-8, VEGF, and NFκB pathways.

Highly Bioavailable Curcumin Derivative 'Theracurmin®' for Crohn's Disease: A Randomized, Double-Blind, Placebo-Controlled Multicentre Study

Background: The new curcumin derivative, Theracurmin® has a 27-fold higher absorption rate than the natural curcumin powder. Its pharmacologic actions include inhibition of the nuclear factor NF-κB, known to mediate the expression of inflammatory cytokines. We were interested to evaluate efficacy of Theracurmin® in patients with Crohn's disease (CD). Methods: Theracurmin® (360mg/day, n=20) or placebo (n=10) was administered to patients with active mild to moderate CD for 12 weeks. Efficacy assessment included clinical and endoscopic remission, healing of anal lesions, and blood levels of inflammatory markers. Findings: In the Theracurmin® group, significant reduction of clinical disease activity was observed at 12 weeks relative to week 0 (P=0.005). The clinical remission rates were 35%, 40%, and 40% at weeks 4, 8, and 12, respectively, significantly higher than in the placebo group (P=0.033, P=0.020, and P=0.020). Further, in the Theracurmin® group, significant healing of anal lesions (P=0.017) and endoscopic CD severity (P=0.032) were observed at weeks 8 and 12, respectively, relative to week 0. However, the mucosal healing rate at week 12 in the Theracurmin® group was 15.0%. No serious adverse event was observed in either group throughout our observation time. Interpretation: In this study, Theracurmin® in patients with active mild to moderate CD produced significant clinical and endoscopic efficacy together with a favourable safety profile. Trial registration: UMI15770 Funding Statement: Theracurmin® was a gift from Theravalues Corporation, Tokyo, Japan. The authors received no external funding for this study. Declaration of Interests: All authors acknowledge having no conflict of interest in connection with the publication of this manuscript. Ethics Approval Statement: At each institution, the study protocol was reviewed and approved by the committee on the Ethics of Clinical Trials involving human subjects. Further, the trial was conducted by adherence to the principles of Good Clinical Practice (GCP) and the ethical standards laid down in the 1964 Helsinki Declaration and its subsequent amendments. Similarly, informed consent was obtained from all patients after explaining the purpose of the study and the nature of the procedures involved.

Targeting MIAT reduces apoptosis of cardiomyocytes after ischemia/reperfusion injury

ABSTRACT This study aims to investigate the role of targeting lncRNA myocardial infarction-associated transcript (MIAT) in protection against hypoxia/reoxygenation (H/R) injury in H9c2 cells in vitro and myocardial ischemia/reperfusion (I/R) injury in vivo by regulating expression of NF-kB and p53 upregulated modulator of apoptosis (PUMA). H9C2 cells were infected with lentivirus expressing the short-hairpin RNA direct against human MIAT gene (Lv-MIAT shRNA) or lentivirus expressing scrambled control (Lv-NC shRNA) or PUMA siRNA or p65 siRNA or their control siRNA respectively. Then the H9c2 cells were infected with Lv-shRNA to 2 hours of hypoxia (H) and 24 hour of reoxygenation (R). 100 ul of Lv-MIAT shRNA (1 × 108 PFU) or Lv-NC shRNA was transfected into mouse hearts, then the hearts were subjected to I/R (1h/72 h). We discovered targeting MIAT remarkably enhanced H9c2 cell viability, decreased H/R-induced cell apoptosis and LDH leakage and significantly decreased I/R-induced myocardial infarct size, reduced myocardial apoptosis and enhanced the heart function. Targeting MIAT downregulated p65 nuclear translocation, NF-κB activity and anti-apoptotic protein cleaved-caspase-3, Bax, and upregulated anti-apoptotic protein Bcl-2 induced by H/R or I/R. Our study suggests that targeting MIAT may protect against H9c2 cardiomyoblasts H/R injury or myocardial I/R injury via inhibition of cell apoptosis, mediated by NF-κB and PUMA signal pathway.

Role of Signaling Molecules in the Regulation of Granulocytopoiesis during Stress-Inducing Stimulation.

The role of signaling molecules in synthesis of humoral regulators of granulocytopoiesis by the hematopoietic microenvironmental cells during stress was analyzed using specific inhibitors. The major role in stimulation of the synthesis of granulocytic CSF during stressful stimulation is played by PI3K/Akt signaling cascade. Nuclear transcription factor NF-κB plays an auxiliary role in the regulation of functional activity of the bone marrow mononuclears. However, this factor affects the synthesis of granulocytic CSF by CD4+ cells of the bone marrow in response to stressful stimulation. Different degree and specific character of involvement of the signaling proteins in the regulation of the production of humoral factors determining colony-stimulating activity are explained by changes in functional state of monocyte-derived macrophages in different periods of stress response.

Hydrogen sulfide treatment alleviated ventilator-induced lung injury through regulation of autophagy and endoplasmic reticulum stress.

Mechanical ventilation has significant therapeutic benefits, but it may cause or aggravate lung injury, which is called ventilator-induced lung injury (VILI). Endogenous hydrogen sulfide (H2S) has roles including regulating inflammation, and promoting vasodilatation; it also exhibits anti-oxidative stress and anti-fibrosis effects. H2S has been reported to alleviate lung injury, but the effects and mechanism of H2S on VILI remain unclear. The present study established a rat model of VILI and treated them with H2S, then measured the changes in respiratory function indicators, lung tissue histopathology, and oxidative, inflammatory, and apoptotic indicators. The effect of H2S on autophagy in the VILI model and the involvement of endoplasmic reticulum (ER) stress were also investigated. To further explore the mechanism, L2 alveolar epithelial cells were treated with cyclic strain to mimic mechanical strain along with the H2S donor NaHS, and the involvement of the NF-κB/MAPK signaling pathway was examined. The results showed that H2S significantly alleviated VILI and inhibited the inflammation and oxidative stress induced by VILI. H2S also significantly reduced autophagy and ER stress in rats. The phosphorylation of IRE1α, PERK and eIF2α and the expression of nuclear ATF4, and GADD34 in L2 cells were all significantly reduced with NaHS. Nuclear NF-κB p65, MAPK p38, JNK, and ERK were all activated by cyclic strain, but inhibited by the ER stress inhibitor 4-PBA or NaHS. Our findings revealed that H2S treatment alleviated VILI by regulating autophagy and ER stress, and the PERK/eIF2α/ATF4/GADD34 and NF-κB/MAPK pathways were involved in the underlying mechanism.