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Background: Natural products have been recommended as a complementary therapy for type 2 diabetes mellitus (T2DM) due to constraints of safety and tolerability of existing anti-diabetic agents. Luteolin exhibits anti-diabetic and anti-inflammatory effects. Hence, the impact of luteolin on glucose homoeostasis and organ damage was investigated in high-fat diet (HFD) and streptozotocin (STZ) induced T2DM in rats. Methods and results: Male Wistar rats were maintained on HFD (provided 55% energy as fat) for 10 days. Subsequently, a single dose of 40 mg/kg STZ was injected intraperitoneally on the 11th day. Seventy-two hours after STZ administration, diabetic rats with established hyperglycemia (fasting serum glucose > 200 mg/dL) were randomized into different groups having six rats each and orally administered either 0.5% hydroxy propyl cellulose or pioglitazone (10 mg/kg) or luteolin (50 mg/kg or 100 mg/kg) once daily for 28 days, while continuing HFD for respective groups. Luteolin significantly reduced hyperglycaemia, homoeostasis model assessment (HOMA) of insulin resistance (HOMA-IR) levels, and improved hypoinsulinemia and HOMA of b-cell function (HOMA-B) in a dose-dependent manner. Increased TNF-α, IL-6 and NFκB levels in diabetic rats were significantly regulated. Additionally, luteolin significantly augmented PPAR-γ expression while attenuating sterol regulatory element binding protein-1c (SREBP-1c) expression. Histopathological scrutiny validated that luteolin effectively attenuated HFD-STZ-induced injury in pancreatic β-cells and kidneys to near normalcy. Conclusion: Our study showed that luteolin ameliorated hyperglycemia and improved hypoinsulinemia, β-cell dysfunction, and renal impairment in HFD-STZ-induced diabetic rats by attenuating inflammation and dysregulated cytokine secretion through modulation of PPAR-γ, TNF-α, IL-6 and NF-kB expression and down-regulation of SREBP-1c. [ABSTRACT FROM AUTHOR]

Objective To observe the preventive and therapeutic effect of intraperitoneal and subcutaneous injection of allopregnanolone (Allo) on postoperative delirium (POD) in aged male mice and to explore its possible mechanism. Methods Ninety-six 6-month-old male C57BL/6J mice were randomly divided into groups A, B, C and control group, with 24 in each. Mice in the control group were fed normally, and mice in the groups A, B and C underwent internal fixation of tibial fracture of right lower limb (POD animal model preparation method is commonly used). Mice in the group A were intraperitoneally injected with 8 mg/kg allopregnenolone at 1 h after surgery, and then were subcutaneously injected with 8 mg/kg allopregnenolone at 6, 24 and 48 h after surgery, respectively. Mice in the group B were intraperitoneally injected with 2-hydroxypropyl-β-cyclodextrin at 1 h after surgery, and subcutaneously injected with 2-hydroxypropyl-β-cyclodextrin at 6, 24 and 48 h after surgery, respectively. Mice in the group C were not given any drugs after surgery. Morris water maze test was used to evaluate the spatial learning and memory ability of mice before surgery and at the 1, 3 and 7 days after surgery, respectively. After the test, six mice were randomly killed in each group. Hippocampal tissues were taken to detect inflammatory factors and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK), nuclear factor κB (NF-κB), and NOD-like receptor protein 3 (NLRP3) pathway-related proteins in the hippocampal tissues of mice in each group. Results Compared with the control group, the escape latency of mice in group C was longer and the number of platform crossing was smaller on the 1st, 3rd and 7th days after operation (all P<0. 05). Compared with the group C, the escape latency of mice in the group A was shorter and the number of platform crossing was lager on day 1, 3 and 7 after operation (all P<0. 05). Compared with the control group, the expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), p-p38MAPK, NF-κB, and NLRP3 in the hippocampal tissues of group C were higher on day 1, 3 and 7 after surgery (all P<0. 05). Compared with the group C, the expression levels of TNF-α, IL-1β, p-p38MAPK, NF-κB and NLRP3 in the hippocampal tissues of the group A were lower on day 1, 3 and 7 after surgery (all P<0. 05). Conclusion Intraperitoneal and subcutaneous injection of allopregnenolone can reduce POD after internal fixation of tibial fracture of right lower limb in aged male mice probably by inhibiting the expression levels of TNF-α, IL-1β, p-p38MAPK, NF-κB, and NLRP3 in the hippocampal tissues. [ABSTRACT FROM AUTHOR]

Hypoxia-inducible factor-1 alpha (HIF-1α) is usually regarded as a core regulator of hypoxic response. Persistent inflammation and impaired wound healing are common manifestations of diabetic foot ulcer (DFU). In normal wounds, HIF-1α and its related regulatory molecules, such as vascular endothelial growth factor and inducible nitric oxide synthase, are activated by hypoxia signals, which in turn promote wound healing. However, abnormal regulation of the HIF-1α signaling pathway by hyperglycemia leads to impaired wound healing in DFU. In this review, we highlight the tissue-specific and stage-specific effects of the HIF-1α signaling pathway in DFU. In the early stage of DFU, HIF-1α in inflammatory cells is over-upregulated by hyperglycemia, causing the activation of nuclear factor-κB and the inducible nitric oxide synthase-mediated pro-inflammatory signaling pathway, leading to sustained inflammation, which is deleterious. In the late stage of DFU, HIF-1α in endothelial cells and keratinocytes is inhibited by hyperglycemia, which leads to the downregulation of vascular endothelial growth factor expression, resulting in insufficient angiogenesis and difficult healing at the wound site. In this review, we discuss recent advances in the knowledge of the HIF-1α signaling pathway and the key targeted molecules in impaired wound healing of DFU. We also summarize the drugs currently in clinical trials that target HIF-1α or its downstream molecules, recapitulate current gaps in our knowledge, and propose rational therapeutic strategies for DFU based on the action characteristics of HIF-1α. [ABSTRACT FROM AUTHOR]

BACKGROUND: Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia, excessive release of inflammatory factors and redox imbalance. However, the specific mechanism is not fully understood. OBJECTIVE: To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS: Passage 3 BV2 microglia were divided into three groups: control group was cultured for 24 hours (without any treatment). Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours. Lipopolysaccharide + nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours. After the processing, cell proliferation was detected by CCK-8 assay. The level of intracellular reactive oxygen species was detected by fluorescent probe. The mRNA expression levels of nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β were detected by qRT-PCR. The protein expression levels of nuclear factor κB p65, p-nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, the proliferation activity of lipopolysaccharide group was decreased (P < 0.001). Compared with the lipopolysaccharide group, the cell proliferation activity of lipopolysaccharide + nobiletin group was increased (P < 0.001). Compared with the control group, the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group (P < 0.001). Compared with the lipopolysaccharide group, the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide + nobiletin group (P < 0.01). Compared with the control group, the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group (P < 0.001, P < 0.01). Compared with the lipopolysaccharide group, mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide + nobiletin group (P < 0.01, P < 0.05). Compared with the control group, the protein expression levels of p-nuclear factor κB p65, tumor necrosis factor α, and interleukin1β in were increased the lipopolysaccharide group (P < 0.001). Compared with the lipopolysaccharide group, the expression of p-nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β was decreased in the lipopolysaccharide + nobiletin group (P < 0.001). These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression levels and significance of serum interleukin-17 （IL-17） and nuclear factor-κB （NF-κB） in Mycoplasma pneumoniae pneumonia （MPP） children in acute stage with lung consolidation. Methods Sixty-four children diagnosed with MPP in acute stage were selected. According to the imaging features of chest CT scan or chest X-ray upon admission， all patients were divided into the lung consolidation group （n = 32） and non-lung consolidation group （n = 32）. The serum levels of IL-17 and NF-κB in MPP children were detected and compared between two groups. The relationship between IL-17 and NF-κB was analyzed. The receiver operating characteristic （ROC） curve was drawn. The diagnostic efficacy of serum IL-17 and NF-κB levels for lung consolidation was assessed in MPP children in acute stage. Results The serum levels of IL-17 and NF-κB in the lung consolidation group were higher than those in the non-lung consolidation group （both P < 0.05）. Spearman correlation analysis showed that IL-17 level was positively correlated with NF-κB level in MPP children （rs = 0.818， P < 0.05）. When the serum level of IL-17 was 10.45 pg/mL， the area under the ROC curve （AUC） was 0.806， and the sensitivity and specificity for the diagnosis of lung consolidation in MPP children were 75.0% and 78.1%. When the serum level of NF-κB was 534.13 pg/mL， the AUC was 0.951， and the sensitivity and specificity for the diagnosis of lung consolidation in children with MPP were 90.6% and 84.4%. Conclusions The expression levels of IL-17 and NF-κB are up-regulated in MPP children with lung consolidation in acute stage. Early detection of serum levels of IL-17 and NF-κB has certain diagnostic efficacy for lung consolidation in MPP.

The key regulators of the transcriptional response to hypoxia and inflammation (hypoxia inducible factor, HIF, and nuclear factor-kappa B, NF-κB, respectively) are evolutionarily conserved and share significant crosstalk. Tissues often experience hypoxia and inflammation concurrently at the site of infection or injury due to fluid retention and immune cell recruitment that ultimately reduces the rate of oxygen delivery to tissues. Inflammation can induce activity of HIF-pathway genes, and hypoxia may modulate inflammatory signaling. While it is clear that these molecular pathways function in concert, the physiological consequences of hypoxia-induced inflammation and how hypoxia modulates inflammatory signaling and immune function are not well established. In this review, we summarize known mechanisms of HIF and NF-κB crosstalk and highlight the physiological consequences that can arise from maladaptive hypoxia-induced inflammation. Finally, we discuss what can be learned about adaptive regulation of inflammation under chronic hypoxia by examining adaptive and maladaptive inflammatory phenotypes observed in human populations at high altitude. We aim to provide insight into the time domains of hypoxia-induced inflammation and highlight the importance of hypoxia-induced inflammatory sensitization in immune function, pathologies, and environmental adaptation.

BACKGROUND: Astrocytes can be activated into A1 and A2 subtypes of reactive astrocytes, which are completely different in gene, molecule and function, and have different specific roles in spinal cord injury repair. High-mobility group box 1 (HMGB1) has been proven to be an important inflammatory and immune factor after spinal cord injury, which can cause cell edema, inflammatory reaction and apoptosis; however, its effect on reactive astrocytes remains unclear. OBJECTIVE: To investigate the activation of rat spinal cord astrocytes into reactive astrocytes after oxygen-glucose deprivation/restoration (OGD/R) and to explore the effect of HMGB1 on different subtypes of reactive astrocytes and the relevant mechanism. METHODS: Rat spinal cord astrocytes were cultured to the third generation. Cell morphology was observed after OGD/R treatment, activation of reactive astrocytes was detected by western blot assay, and migration ability of the cells was detected by cell scratch assay. After inhibition of HMGB1, the cells were divided into five groups: normal group, OGD6 h/R6 h group, HMGB1 interference group, negative sequence group, and OGD6 h/R6 h+12 μmol/L ethyl pyruvate group (an HMGB1 inhibitor). Western blot assay and immunofluorescence method were used to detect the expression of C3 and S100A10 in different subtypes of reactive astrocytes, and cell scratch assay was used to detect the migration of reactive astrocytes. To explore the possible mechanism underlying the effect of HMGB1 on reactive astrocytes, the cells were divided into four groups: normal group, OGD6 h/R6 h group, OGD6 h/R6 h+5 µmol/L CLI-095 group (a toll-like receptor 4 inhibitor), and OGD6 h/R6 h+5 µmol/L BAY 11-7082 group (a nuclear factor-κB inhibitor). Western blot assay was used to detect the expression of toll-like receptor 4, nuclear factor-B, C3, and S100A10. RESULTS AND CONCLUSION: (1) Oxygen-glucose deprived cells were shrunk in size and had an increased migration capacity. After reoxygen-glucose treatment, the cell volume was increased and A1 and A2 reactive astrocytes were activated. S100A10 expression was the highest at OGD6 h/R6 h and C3 expression was peaked at OGD6 h/R12 h (P < 0.05). (2) Effects of silencing or inhibition of HMGB1 expression on reactive astrocytes: Compared with the OGD6 h/R6 h group, inhibition of HMGB1 significantly reduced the expression of C3 (P < 0.05), increased the expression of S100A10 (P < 0.05), and enhanced cell migration ability. (3) Compared with the OGD6 h/R6 h group, the expression of toll-like receptor 4 decreased in the OGD6 h/R6 h+CLI 095 group (P < 0.05), while the expression of nuclear factor-κB and C3 decreased and the expression of S100A10 increased in the OGD6 h/R6 h+CLI 095 group and OGD6 h/R6 h+BAY 11-7082 group (P < 0.05). (4) To conclude, astrocytes can be activated into two different subtypes of reactive astrocytes after OGD/R. Inhibition of HMGB1 can reduce A1 reactive astrocytes, increase A2 reactive astrocytes, and enhance cell migration ability through the toll-like receptor 4/nuclear factor-κB pathway. [ABSTRACT FROM AUTHOR]

Introduction: Lipopolysaccharide (LPS) tolerance is the downregulation of LPS signaling after pre-exposure to LPS, and it provides protection against hyperactive inflammation. Cytokine production decreases during LPS tolerance, and the phenotype of LPS-tolerant monocytes shifts toward M2 (anti-inflammatory) type. Magnesium sulfate (MgSO4) is a widely used anti-inflammatory agent. Although MgSO4 inhibits LPS signaling, the effect of MgSO4 on LPS tolerance is unknown. In the present study, we investigated the in vitro effects of MgSO4 on LPS tolerance. Methods: To induce LPS tolerance, THP-1 cells were stimulated with LPS (200 ng/mL, 2 h) after pre-exposure to LPS (200 ng/mL, 24 h) with or without pre-treatment of MgSO4 (20 mM, 24 h). Results: Our results revealed that MgSO4 enhanced LPS tolerance by downregulating nuclear factor-κB (NF-κB)-induced tumor necrosis factor-α or interleukin-6, and upregulating cluster of differentiation 163 (a M2-associated marker). Furthermore, the LPS-triggered upregulation of phosphoinositide 3-kinase (PI3K) was significantly increased during LPS tolerance. MgSO4 activated PI3K, but inhibited NF-κB in LPS-stimulated cells. Notably, MgSO4 mitigated the signaling of both PI3K and NF-κB in LPS-tolerant cells, suggesting the effect of MgSO4 on LPS tolerance relies on the modulation of the crosstalk between PI3K and NF-κB. Conclusion: MgSO4 enhanced LSP tolerance, thus providing evidence for a novel underlying mechanism of the anti-inflammatory effects of MgSO4. [ABSTRACT FROM AUTHOR]

Purpose: BK Polyomavirus (BKPyV) infection manifests as renal inflammation and can cause kidney damage. Tumor necrosis factor-α (TNF-α) is increased in renal inflammation and injury. The aim of this study was to investigate the effect of TNF-α blockade on BKPyV infection. Methods: Urine specimens from 22 patients with BKPyV-associated nephropathy (BKPyVN) and 35 non-BKPyVN kidney transplant recipients were analyzed. Results: We demonstrated increased urinary levels of TNF-α and its receptors, TNFR1 and TNFR2, in BKPyVN patients. Treating BKPyV-infected human proximal tubular cells (HRPTECs) with TNF-α stimulated the expression of large T antigen and viral capsid protein-1 mRNA and proteins and BKPyV promoter activity. Knockdown of TNFR1 or TNFR2 expression caused a reduction in TNF-α-stimulated viral replication. NF-κB activation induced by overexpression of constitutively active IKK2 significantly increased viral replication and the activity of the BKPyV promoter containing an NF-κB binding site. The addition of a NF-κB inhibitor on BKPyV-infected cells suppressed viral replication. Blockade of TNF-α functionality by etanercept reduced BKPyV-stimulated expression of TNF-α, interleukin-1β (IL-1β), IL-6 and IL-8 and suppressed TNF-α-stimulated viral replication. In cultured HRPTECs and THP-1 cells, BKPyV infection led to increased expression of TNF-α, interleukin-1 β (IL-1β), IL-6 and TNFR1 and TNFR2 but the stimulated magnitude was far less than that induced by poly(I:C). This may suggest that BKPyV-mediated autocrine effect is not a major source of TNFα. Conclusion: TNF-α stimulates BKPyV replication and inhibition of its signal cascade or functionality attenuates its stimulatory effect. Our study provides a therapeutic anti-BKPyV target. [ABSTRACT FROM AUTHOR]

Inflammatory processes play major roles in carcinogenesis and the progression of hepatocellular carcinoma (HCC) derived from non-alcoholic steatohepatitis (NASH). But, there are no therapies for NASH-related HCC, especially focusing on these critical steps. Previous studies have reported that farnesyltransferase inhibitors (FTIs) have anti-inflammatory and anti-tumor effects. However, the influence of FTIs on NASH-related HCC has not been elucidated. In hepatoblastoma and HCC cell lines, HepG2, Hep3B, and Huh-7, we confirmed the expression of hypoxia-inducible factor (HIF)-1α, an accelerator of tumor aggressiveness and the inflammatory response. We established NASH-related HCC models under inflammation and free fatty acid burden and confirmed that HIF-1α expression was increased under both conditions. Tipifarnib, which is an FTI, strongly suppressed increased HIF-1α, inhibited cell proliferation, and induced apoptosis. Simultaneously, intracellular interleukin-6 as an inflammation marker was increased under both conditions and significantly suppressed by tipifarnib. Additionally, tipifarnib suppressed the expression of phosphorylated nuclear factor-κB and transforming growth factor-β. Finally, in a NASH-related HCC mouse model burdened with diethylnitrosamine and a high-fat diet, tipifarnib significantly reduced tumor nodule formation in association with decreased serum interleukin-6. In conclusion, tipifarnib has anti-tumor and anti-inflammatory effects in a NASH-related HCC model and may be a promising new agent to treat this disease. [ABSTRACT FROM AUTHOR]

Turmeronols (A and B), bisabolane-type sesquiterpenoids found in turmeric, reduce inflammation outside the brain in animals; however, their effects on neuroinflammation, a common pathology of various neurodegenerative diseases, are not understood. Inflammatory mediators produced by microglial cells play a key role in neuroinflammation, so this study evaluated the anti-inflammatory effects of turmeronols in BV-2 microglial cells stimulated with lipopolysaccharide (LPS). Pretreatment with turmeronol A or B significantly inhibited LPSinduced nitric oxide (NO) production; mRNA expression of inducible NO synthase; production of interleukin (IL)-1β, IL-6, and tumor necrosis factor α and upregulation of their mRNA expression; phosphorylation of nuclear factor-κB (NF-κB) p65 proteins and inhibitor of NF-κB kinase (IKK); and nuclear translocation of NF-κB. These results suggest that these turmeronols may prevent the production of inflammatory mediators by inhibiting the IKK/NF-κB signaling pathway in activated microglial cells and can potentially treat neuroinflammation associated with microglial activation. [ABSTRACT FROM AUTHOR]

Linear ubiquitin chain assembly complex (LUBAC) is a ubiquitin E3 ligase complex composed of HOIP, HOIL-1L, and SHARPIN that catalyzes the formation of linear/M1- linked ubiquitin chain. It has been shown to play a pivotal role in the nuclear factor (NF)- κB signaling induced by proinflammatory stimuli. Here, we found that tumor susceptibility gene (TSG101) physically interacts with HOIP, a catalytic component of LUBAC, and potentiates LUBAC activity. Depletion of TSG101 expression by RNA interference decreased TNFa-induced linear ubiquitination and the formation of TNFa receptor 1 signaling complex (TNF- RSC). Furthermore, TSG101 facilitated the TNFa-induced stimulation of the NF-κB pathway. Thus, we suggest that TSG101 functions as a positive modulator of HOIP that mediates TNFa-induced NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

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Objective To investigate the expression of MicroRNA-27a (miR-27a) in the placenta of gestational diabetes mellitus (GDM) rats and its effects on oxidative stress and inflammation in GDM rats, as well as to analyze the underlying mechanism. Methods SD normal pregnant rats were injected intraperitoneally with strepto-zotocin (STZ) to induce the establishment of GDM models. After successful modeling, they were divided into four groups: model group, agomir-NC group, miR-27a agomir group, and miR-27a agomir + lipopolysaccharide (LPS) group, each with 12 rats; the control group consisted of12 normal pregnant rats. The levels of fasting blood glucose (FBG) and fasting insulin (FINS) in rats were detected, and the insulin resistance index (HOMA-IR) was calculated; the pregnancy outcome and placenta quality were analyzed; the levels of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α), the activities of superoxide dismutase (SOD) and catalase (CAT), and the content of malondialdehyde (MDA) in placenta tissue were determined; Hematoxylin-Eosin (HE) staining and TUNEL method were used to detect pancreatic and placental tissue damage and cell apoptosis; and real -time quantitative PCR (RT - qPCR) and Western blot were used to detect the levels of miR-27a, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor-κB p65 (NF-κB p65) mRNAs and proteins in placental tissues. Results Overexpression of miR-27a significantly reduced FBG, FINS and HOMA-IR in GDM rats, accompanied by a reduction in pro-inflammatory cytokine levels and oxidative stress, improved placental and pancreatic tissue damage and apoptosis, and the pregnancy outcome, and inhibited the activation of the TLR4/MyD88/NF -κB signaling pathway in the placenta (P < 0.05). Conclusion The overexpression of miR -27a can reduce the placental inflammation, oxidative stress, and IRin GDM rats. Its mechanism could be related to the inhibition of the TLR4/MyD88/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

The c-Jun N-terminal kinase (JNK) family includes three proteins (JNK1-3) that regulate many physiological processes, including cell proliferation and differentiation, cell survival, and inflammation. Because of emerging data suggesting that JNK3 may play an important role in neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease, as well as cancer pathogenesis, we sought to identify JNK inhibitors with increased selectivity for JNK3. A panel of 26 novel tryptanthrin-6-oxime analogs was synthesized and evaluated for JNK1-3 binding (Kd) and inhibition of cellular inflammatory responses. Compounds 4d (8-methoxyindolo[2,1-b]quinazolin-6,12-dione oxime) and 4e (8-phenylindolo[2,1-b]quinazolin-6,12-dione oxime) had high selectivity for JNK3 versus JNK1 and JNK2 and inhibited lipopolysaccharide (LPS)-induced nuclear factor-κB/activating protein 1 (NF-κB/AP-1) transcriptional activity in THP-1Blue cells and interleukin-6 (IL-6) production by MonoMac-6 monocytic cells in the low micromolar range. Likewise, compounds 4d, 4e, and pan-JNK inhibitor 4h (9-methylindolo[2,1-b]quinazolin-6,12-dione oxime) decreased LPS-induced c-Jun phosphorylation in MonoMac-6 cells, directly confirming JNK inhibition. Molecular modeling suggested modes of binding interaction of these compounds in the JNK3 catalytic site that were in agreement with the experimental data on JNK3 binding. Our results demonstrate the potential for developing anti-inflammatory drugs based on these nitrogen-containing heterocyclic systems with selectivity for JNK3. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases (CVDs) are the leading cause of death globally. Atherosclerosis is the basis of major CVDs – myocardial ischemia, heart failure, and stroke. Among numerous functional molecules, the transcription factor nuclear factor κB (NF-κB) has been linked to downstream target genes involved in atherosclerosis. The activation of the NF-κB family and its downstream target genes in response to environmental and cellular stress, hypoxia, and ischemia initiate different pathological events such as innate and adaptive immunity, and cell survival, differentiation, and proliferation. Thus, NF-κB is a potential therapeutic target in the treatment of atherosclerosis and related CVDs. Several biologics and small molecules as well as peptide/proteins have been shown to regulate NF-κB dependent signaling pathways. In this review, we will focus on the function of NF-κB in CVDs and the role of NF-κB inhibitors in the treatment of CVDs. [ABSTRACT FROM AUTHOR]

Low doses of antibiotics are ubiquitous in the marine environment and may exert negative effects on non-target aquatic organisms. Using primary macrophages of common carp, we investigated the mechanisms of action following exposure to several common antibiotics; cefotaxime, enrofloxacin, tetracycline, sulfamonomethoxine, and their mixtures, and explored the immunomodulatory effects associated with the nuclear factor-κB (NF-κB) signaling pathway. A KEGG pathway analysis was conducted using the sixty-six differentially expressed genes found in all treatments, and showed that exposure to 100 μg/L of antibiotics could affect regulation of the NF-κB signaling pathway, suggesting that activation of NF-κB is a common response in all four classes of antibiotics. In addition, the four antibiotics induced nf-κb and NF-κB-associated cytokines expression, as verified by qPCR, however, these induction responses by four antibiotics were minor when compared to the same concentration of LPS treatment (100 μg/L). Antagonists of NF-κB blocked many of the immune effects of the antibiotics, providing evidence that NF-κB pathways mediate the actions of all four antibiotics. Moreover, exposure to environmentally relevant, low levels (0.01-100 μg/L) of antibiotics induced a NF-κB-mediated immune response, including endogenous generation of ROS, activity of antioxidant enzymes, as well as expression of cytokine and apoptosis. Moreover, exposure to mixtures of antibiotics presented greater effects on most tested immunological parameters than exposure to a single antibiotic, suggesting additive effects from multiple antibiotics in the environment. This study demonstrates that exposure of fish primary macrophages to low doses of antibiotics activates the NF-kB pathway.

BackgroundRadiation-induced liver damage is a major complication for primary liver cancer and other upper abdominal tumors during radiation therapy. The early biological effects of radiation-induced liver damage at different doses of radiation and its mechanisms of action have not yet been elucidated. ObjectiveTo establish X-ray-induced radioactive mouse liver damage model and explore the level of oxidative stress and its correlation with nuclear factor-κB (NF-κB) and transforming growth factor-β1 (TGF-β1). MethodsA total of 24 male C57BL/6J mice aged 6 weeks were randomly divided into 4 groups (control, 0.8 Gy, 1.6 Gy, and 4 Gy), with 6 mice in each group. X-rays irradiated the whole body of mice singly in each dose group. At 24 h after radiation, histopathological changes in mouse liver were evaluated; peripheral blood cell count, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, as well as liver tissue superoxide dismutase (SOD) activity, malondialdehyde (MDA) level, reduced glutathione (GSH) level, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) level were measured; real-time fluorescence quantitative PCR was used to detect liver tissue NF-κB p65 and TGF-β1 mRNA expression levels; the correlations of oxidative stress indicators with NF-κB p65 and TGF-β1 mRNA expression levels were analyzed by Pearson correlation. ResultsCompared with the control group, at 24 h after different doses of X-ray radiation, early injury-related histopathological changes were observed in liver, and the serum levels of AST and ALT were significantly increased in the 4 Gy group (P

ABSTRACT: Excessive inflammation in bovine mammary endothelial cells (BMEC) due to mastitis leads to disease progression and eventual culling of cattle. Sirtuin 3 (SIRT3), a mitochondrial deacetylase, downregulates pro-inflammatory cytokines in BMEC exposed to high concentrations of nonesterified fatty acids by blunting nuclear factor-κB (NFκB) signaling. In nonruminants, SIRT3 is under the control of PGC1α, a transcriptional cofactor. Specific aims were to study (1) the effect of SIRT3 on inflammatory responses of lipopolysaccharide (LPS)-challenged bovine mammary epithelial cells (bovine mammary alveolar cells-T, MAC-T) models, and (2) the role of PGC1α in the attenuation of NFκB signaling via SIRT3. To address these objectives, first, MAC-T cells were incubated in triplicate with 0, 50, 100, 150, or 200 μg/mL LPS (derived from Escherichia coli O55:B5) for 12 h with or without a 2-h incubation of the NFκB inhibitor ammonium pyrrolidine dithiocarbamate (APDC, 10 μM). Second, SIRT3 was overexpressed using adenoviral expression (Ad-SIRT3) at different multiplicity of infection (MOI) for 6 h followed by a 12 h incubation with 150 μg/mL LPS. Third, cells were treated with the PGC1α agonist ZLN005 (10 μg/mL) for 24 h and then challenged with 150 μg/mL LPS for 12 h. Fourth, cells were initially treated with the PGC1α inhibitor SR-18292 (100 μM) for 6 h followed by a 6-h culture with or without 50 MOI Ad-SIRT3 and a challenge with 150 μg/mL LPS for 12 h. Data were analyzed using one-way ANOVA with subsequent Bonferroni correction. Linear and quadratic contrasts were used to determine dose-responses to LPS. There were linear and quadratic effects of LPS dosage on cell viability. Incubation with 150 and 200 μg/mL LPS for 12 h decreased cell viability to 78.6 and 34.9%, respectively. Compared with controls, expression of IL1B, IL6, and TNFA was upregulated by 5.2-, 5.9-, and 2.7-fold with 150 μg/mL LPS; concentrations of IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) in cell medium also increased. Compared with the LPS group, LPS+APDC increased cell viability and reversed the upregulation of IL1B, IL6, and TNFA expression. However, mRNA and protein abundance of SIRT3 decreased linearly with increasing LPS dose. Ad-SIRT3 infection (50 MOI) reduced IL1B, IL6, and TNFA expression and also their concentrations in cell medium, and decreased pNFκB P65/NFκB P65 ratio and nuclear abundance of NFκB P65. The PGC1α agonist increased SIRT3 expression, whereas it decreased cytokine expression, pNFκB P65/NFκB P65 ratio, and prevented NFκB P65 nuclear translocation. Contrary to the agonist, the PGC1α inhibitor had opposite effects, and elevated the concentrations of IL-1β, IL-6, and TNF-α in cell medium. Overall, data suggested that SIRT3 activity could attenuate LPS-induced inflammatory responses in mammary cells via alterations in the PGC1α-NFκB pathway. As such, there may be potential benefits for targeting SIRT3 in vivo to help prevent or alleviate negative effects of mastitis.

Nasopharyngeal carcinoma (NPC) is a prevalent malignant tumor worldwide. FKBP3 has been reported to participate in tumorigenesis. Nevertheless, the role and mechanism of FKBP3 in NPC remains unclear. In this study, FKBP3 expression was observed to upregulate in NPC patients and cells. Moreover, knockdown of FKBP3 suppressed cell growth, invasion, and migration in HK1 and C666-1 cells. Mechanically, FKBP3 could enhance the p-p65 expression and activated p65 signaling pathway and increased interleukin-6 (IL-6) expression through enhancing histone deacetylase 2 (HDAC2) expression. In rescued experiment, the overexpression of HDAC2 restored diminished cell growth, invasion, and migration caused by FKBP3 depletion. In summary, the knockdown of FKBP3 suppressed NPC cell growth, invasion and migration, deactivated nuclear factor-κB/IL-6 signaling pathway through inhibiting HDAC2 expression, providing a potential therapeutic strategy for NPC treatment.

Abstract Background Inflammation is present in neurological and peripheral disorders. Thus, targeting inflammation has emerged as a viable option for treating these disorders. Previous work indicated pretreatment with beta-funaltrexamine (β-FNA), a selective mu-opioid receptor (MOR) antagonist, inhibited inflammatory signaling in vitro in human astroglial cells, as well as lipopolysaccharide (LPS)-induced neuroinflammation and sickness-like-behavior in mice. This study explores the protective effects of β-FNA when treatment occurs 10 h after LPS administration and is the first-ever investigation of the sex-dependent effects of β-FNA on LPS-induced inflammation in the brain and peripheral tissues, including the intestines. Results Male and female C57BL/6J mice were administered LPS followed by treatment with β-FNA-immediately or 10 h post-LPS. Sickness- and anxiety-like behavior were assessed using an open-field test and an elevated-plus-maze test, followed by the collection of whole brain, hippocampus, prefrontal cortex, cerebellum/brain stem, plasma, spleen, liver, large intestine (colon), proximal small intestine, and distal small intestine. Levels of inflammatory chemokines/cytokines (interferon γ-induced-protein, IP-10 (CXCL10); monocyte-chemotactic-protein 1, MCP-1 (CCL2); interleukin-6, IL-6; interleukin-1β, IL-1β; and tumor necrosis factor-alpha, TNF-α) in tissues were measured using an enzyme-linked immunosorbent assay. Western blot analysis was used to assess nuclear factor-kappa B (NF-κB) expression. There were sex-dependent differences in LPS-induced inflammation across brain regions and peripheral tissues. Overall, LPS-induced CXCL10, CCL2, TNF-α, and NF-κB were most effectively downregulated by β-FNA; and β-FNA effects differed across brain regions, peripheral tissues, timing of the dose, and in some instances, in a sex-dependent manner. β-FNA reduced LPS-induced anxiety-like behavior most effectively in female mice. Conclusion These findings provide novel insights into the sex-dependent anti-inflammatory effects of β-FNA and advance this agent as a potential therapeutic option for reducing both neuroinflammation an intestinal inflammation.

Abstract Background SET domain containing 6 (SETD6) has been shown to be upregulated in multiple human cancers and can promote malignant cell survival. However, expression and function of SETD6 in lung adenocarcinoma (LUAD) remains unaddressed. This study aimed to demonstrate the expression pattern, biological roles and potential mechanisms by which SETD6 dysregulation is associated with LUAD. Methods The expression level of SETD6 was evaluated in LUAD clinical specimens and its correlation with clinical parameters were analyzed. In vitro, gain-of-function and loss-of-function experiments were performed to evaluate the effects of SETD6 on cell proliferation, apoptosis, migration, and colony formation of LUAD cell line A549. Western-blot was performed to investigate the involvement of nuclear factor-κB (NF-κB) and nuclear factor erythroid 2–related factor 2 (Nrf2) pathways as downstream signaling of SETD6 in LUAD cells. Results Compared with non-tumorous tissues, SETD6 was overexpressed in tumor tissues, and its overexpression significantly correlates with higher rates of regional lymph node metastasis and poor prognosis in patients with LUAD. In A549 cell line, SETD6 overexpression could promote cell proliferation, migration, colony formation and inhibit cell apoptosis, whereas SETD6 knockdown caused the opposite effects. Furthermore, we demonstrated that the mechanisms underlying the effect of SETD6 on LUAD biological behaviors may be through its interaction with NF-κB and Nrf2 signaling pathways. Conclusions SETD6, which is highly expressed in LUAD tumor tissues, plays an important role in promoting the malignant behaviors of LUAD via likely the NF-κB and Nrf2 signaling pathways.

Objective To explore the preventive and therapeutic effects of gingerol on lipopolysaccharide （LPS）-induced procoagulant and fibrinolytic inhibition in acute respiratory distress syndrome（ARDS）rats and its possible mechanism. Methods Male SD rats were randomly divided into control group, LPS group and different dose of gingerol group（10 m/kg, 20 m/kg, 30 m/kg）. The ARDS model of rats was induced by LPS intratracheal atomization inhalation and treated by intraperitoneal injection of different doses of gingerol. The pathological changes of rat lung were observed by H&E staining, and the degree of lung injury was evaluated. The wet/dry ratio of lung tissue was calculated to evaluate pulmonary edema;the mRNA expression of tissue factor（TF）, and plasminogen activator inhibitor-1（PAI-1） was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction;western blot was used to detect the protein expressions of TF, PAI-1 and nuclear factor-κB p65（p65）, phosphorylation of p65（p-p65）, inhibitor of nuclear factor-κB kinase（IKKβ）and phosphorylation of IKKβ（p-IKKβ）in lung tissues. Results Compared with those in the control group, the pulmonary tissues in the model group were seen with damaged alveolar structures, widened alveolar septa, increased inflammatory cell infiltration and increased lung injury score. The mRNA level of TF and PAI-1 in lung tissue was significantly increased, and the protein expression level of TF, PAI-1, p65 and IKK in lung tissue of group A was significantly increased. Compared to those in the model group, lung tissues in mice of gingerol groups were seen with decreased deconstruction, narrowed alveolar septa, decreased inflammtory cells infiltration and declined lung injury score. In gingerol groups, pulmonary expressions of TF, PAI-1, p-p65, and p-IKKβ protein were all decreased or declined, among which the changes of above indexes in mice with high dose gingerol were more obvious than those in mice with medium and low dose. Conclusion Gingerol dose-dependently ameliorates alveolar hypercoagulation and fibrinolytic inhibition and reduces lung injury for LPS-induced ARDS in rat, and the effects might be associated with the inactivation of NF-κ B signal pathway. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effect of polymyxin B on hepatitis B virus （HBV） transcription and replication. METHODS: The cytotoxicity of polymyxin B in HepG2-NTCP, HepAD38, HepG2. 2. 15, HepG2, Huh-7 and PLC/PRF/5 cells was examined by MTT assay. The HBV-infected HepG2-NTCP cells and HepG2. 2. 15 cells with stable HBV replication were treated with 10, 50 and 100 nmol/L polymyxin B, and the mRNA and DNA levels of HBV in the cells were measured by RT-qPCR. The levels of hepatitis B surface antigen （HBsAg） and hepatitis B e antigen （HBeAg） in the cell supernatants were quantified by ELISA. The regulatory effect of polymyxin B on HBV transcription and replication was clarified. Subsequently, we used transcriptome sequencing to screen nuclear factor-κB （NF-κB） as the effector molecule of polymyxin B regulating HBV transcription and replication, and we determined the regulatory effect of knockdown and overexpression of NF-κB on HBV transcription and replication. We used dual-luciferase reporter assay to determine the effect of NF-κB on the activity of Cp, Xp, Sp1 and Sp2 promoters. Finally, a reverse complementation assay was used to investigate whether the regulation of HBV transcription and replication by polymyxin B is dependent on NF-κB. RESULTS: The MTT assay results showed that polymyxin B did not exhibit significant toxicity in HepG2-NTCP, HepAD38, HepG2. 2. 15, HepG2, Huh-7 and PLC/PRF/5 cells when used at concentrations less than 100 μmol/L. Polymyxin B decreased the HBV RNA, HBV DNA, HBsAg and HBeAg levels in HBV-infected HepG2-NTCP cells and HepG2. 2. 15 cells with stable HBV replication in a concentration-dependent manner. Transcriptome sequencing revealed that polymyxin B significantly decreased the expression level of NF-κB in HBV-infected HepG2-NTCP cells. Furthermore, overexpression of NF-κB significantly increased the levels of HBV RNA, HBV DNA, HBsAg and HBeAg in HBV-infected HepG2- NTCP cells, and the opposite effect was found when NF-κB was knocked down. Moreover, the results of dual-luciferase reporter assay showed that NF-κB enhanced HBV Sp2 promoter activity. The reverse complementation assay revealed that polymyxin B regulates HBV transcription and replication in a manner dependent on NF-κB. CONCLUSION: Polymyxin B inhibits HBV transcription and replication by down-regulating NF-κB and inhibiting HBV Sp2 promoter activity. [ABSTRACT FROM AUTHOR]

Simple Summary: Kirsten rat sarcoma virus (KRAS)-mutant cancers are frequent, metastatic, lethal, and largely undruggable. The aim of this study was to investigate the pathways through which KRAS-mutant cancers foster their growth, thereby unravelling novel therapeutic targets. We show that KRAS-mutant tumors secrete the protein versican, which then drives the activation of NF-κB kinase (IKK) β in a type of host immune cells called macrophages. Following this activation, macrophages fuel the tumor with interleukin (IL)-1β, to close an inflammatory loop through which KRAS-mutant cancers attract host immune cells to the tumor site to accelerate tumor growth and aggressiveness. Importantly, we show that targeting IL-1β and/or versican can be an effective treatment for KRAS-mutant cancers, holding great promise for cancer patients. Kirsten rat sarcoma virus (KRAS)-mutant cancers are frequent, metastatic, lethal, and largely undruggable. While interleukin (IL)-1β and nuclear factor (NF)-κB inhibition hold promise against cancer, untargeted treatments are not effective. Here, we show that human KRAS-mutant cancers are addicted to IL-1β via inflammatory versican signaling to macrophage inhibitor of NF-κB kinase (IKK) β. Human pan-cancer and experimental NF-κB reporter, transcriptome, and proteome screens reveal that KRAS-mutant tumors trigger macrophage IKKβ activation and IL-1β release via secretory versican. Tumor-specific versican silencing and macrophage-restricted IKKβ deletion prevents myeloid NF-κB activation and metastasis. Versican and IKKβ are mutually addicted and/or overexpressed in human cancers and possess diagnostic and prognostic power. Non-oncogene KRAS/IL-1β addiction is abolished by IL-1β and TLR1/2 inhibition, indicating cardinal and actionable roles for versican and IKKβ in metastasis. [ABSTRACT FROM AUTHOR]

Objective To investigate the role and mechanism of dapagliflozin（Dapa）, a sodium glucose cotransporter 2 inhibitor, in acute liver injury. Methods Eight-week-old C57BL6/J mice were given a single intraperitoneal injection of CCl4 to induce acute liver injury. The mice were preventively given 5 mg/kg Dapa by gavage 24 h and 2 h before CCl4 injection, while those in the control group were given an equal volume of solvent gavage. After 24 h, the mice were anesthetized and sacrificed. H＆E staining, plasma biochemistry, RT-q PCR, and Western blotting were used to detect the severity of liver injury and the expressions of macrophage-related genes. Results In the CCl4 group, hepatic infiltration of inflammatory cells increased, and liver and renal functions significantly deteriorated, which was further aggravated by Dapa. CCl4 could promote the expressions of M1 macrophages and fibrosis-related genes in the liver, but reduce those of M2 and antioxidant-related genes, and the latter was further inhibited by Dapa. In addition, the protein expression of arginase 1 decreased and that of SGLT2 increased after Dapa intervention, while NF-κB pathway did not change significantly, suggesting that Dapa might directly affect the energy metabolism homeostasis in the liver and aggravate acute liver injury induced by CCl4. Conclusion Dapa can exacerbate hepatic and renal damage in acute stage of liver injury, inhibit macrophages M2 polarization, and aggravate oxidative stress and inflammatory injury induced by CCl4. [ABSTRACT FROM AUTHOR]

Abnormal placental inflammation has a role in the pathophysiology of preeclampsia. Magnesium sulfate (MgSO4) has anti-inflammatory properties and is a fetal neuroprotective agent. MgSO4 is often used to treat severe preeclampsia; however, the specificmechanisms of action underlyingthistherapeutic effect remain unclear. The objective of this study was to investigate the effects of MgSO4 (270 mg/kg) on placental inflammation in a rat model of lipopolysaccharide (LPS; 1.0 µg/kg)-induced preeclampsia. Compared to normal pregnant rats, LPS-treated pregnant rats had higher blood pressure, proteinuria, and expression of the anti-angiogenic factor sFlt-1 and the pro-inflammatory factors interleukin-1β (IL-1β) and IL-12 in placental tissue. LPS-treated pregnant rats had placental insufficiency, poor fetal outcomes, and significantly decreased expression of the anti-inflammatory factors apolipoprotein E (APOE) and IL-10 in placental tissue. MgSO4 treatment had favorable effects on maternal and fetal outcomes. MgSO4 treatment improved placental function by repressing an exaggerated inflammatory response in the placenta and promoting angiogenesis via the NF-κB pathway. These findings suggest MgSO4 has a potential role in the prevention of preeclampsia and in the treatment of mild and moderate preeclampsia. [ABSTRACT FROM AUTHOR]

Bauhinia championii is a herbal medicine used to treat osteoarthritis (OA) in Chinese traditional medicine. However, the molecular mechanisms underlying the therapeutic effects of this medicinal herb against OA have rarely been reported. Given that it has been established that extracellular matrix metabolism plays an important role in the pathogenesis of OA, the present study focused on the effects and mechanisms of Bauhinia championii in the regulation of extracellular matrix metabolism in chondrocytes induced by IL-1β. Rat chondrocytes were isolated, cultured and identified in vitro. The CCK-8 method was used to detect the cell viability of Bauhinia championii aqueous extract (BCAE)-treated chondrocytes. The chondrocyte inflammatory and degeneration models were induced by 10 ng/mL IL-1β, then chondrocytes were grouped into different groups to evaluate the effect of BCAE on extracellular matrix degradation and the regulation of TLR4/NF-κB signaling pathway. Furthermore, whether the regulatory effect of BCAE on TLR4/NF-κB signaling pathway is related to miRNA-145-5p was also investigated by cell transfection. We found that BCAE promoted chondrocyte viability in a dose- and time-dependent manner. BCAE delayed chondrocyte degeneration induced by IL-1β. BCAE could reduce the degradation of the cartilage extracellular matrix by inhibiting the TLR4/NF-κB signaling pathway. miRNA-145-5p negatively regulated the expression of TLR4 in chondrocytes, while BCAE could upregulate the expression of miRNA-145-5p in chondrocytes induced by IL-1β. These results suggest that BCAE upregulates the expression of miRNA-145-5p to inhibit the TLR4/NF-κB signaling pathway, thereby alleviating the metabolic imbalance of the extracellular matrix and protecting chondrocytes from degeneration.

Background: Cartilage harvest and transplantation is a common surgery using costal, auricular, and septal cartilage for craniofacial reconstruction. However, absorption and warping of the cartilage grafts can occur due to inflammatory factors associated with wound healing. Transcription factor nuclear factor-κB (NF-κB) is activated by the various stimulation such as interleukin-1 (IL-1), and plays a central role in the transactivation of this inflammatory cytokine gene. Inhibition of NF-κB may have anti-inflammatory effects. The aim of this study was to explore the potential of an NF-κB decoy oligodeoxynucleotide (Decoy) as a chondroprotective agent. Materials and Methods: Safe and efficacious concentrations of Decoy were assessed using rabbit nasal septal chondrocytes (rNSChs) and assays for cytotoxicity, proteoglycan (PG) synthesis, and PG turnover were carried out. The efficacious concentration of Decoy determined from the rNSChs was then applied to human nasal septal cartilage (hNSC) in vitro and analyzed for PG turnover, the levels of inflammatory markers, and catabolic enzymes in explant-conditioned culture medium. Results: Over the range of Decoy conditions and concentrations, no inhibition of PG synthesis or cytotoxicity was observed. Decoy at 10 μM effectively inhibited PG degradation in the hNSC explant, prolonging PG half-life by 63% and decreasing matrix metalloprotease 3 (MMP-3) by 70.7% (p = 0.027). Conclusions: Decoy may be considered a novel chondroprotective therapeutic agent in cartilage transplantation due to its ability to inhibit cartilage degradation due to inflammation cytokines.

Excessive inflammation in bovine mammary endothelial cells (BMEC) due to mastitis leads to disease progression and eventual culling of cattle. Sirtuin 3 (SIRT3), a mitochondrial deacetylase, downregulates pro-inflammatory cytokines in BMEC exposed to high concentrations of nonesterified fatty acids by blunting nuclear factor-κB (NFκB) signaling. In nonruminants, SIRT3 is under the control of PGC1α, a transcriptional cofactor. Specific aims were to study (1) the effect of SIRT3 on inflammatory responses of lipopolysaccharide (LPS)-challenged bovine mammary epithelial cells (bovine mammary alveolar cells-T, MAC-T) models, and (2) the role of PGC1α in the attenuation of NFκB signaling via SIRT3. To address these objectives, first, MAC-T cells were incubated in triplicate with 0, 50, 100, 150, or 200 μg/mL LPS (derived from Escherichia coli O55:B5) for 12 h with or without a 2-h incubation of the NFκB inhibitor ammonium pyrrolidine dithiocarbamate (APDC, 10 μ M). Second, SIRT3 was overexpressed using adenoviral expression (Ad-SIRT3) at different multiplicity of infection (MOI) for 6 h followed by a 12 h incubation with 150 μg/mL LPS. Third, cells were treated with the PGC1α agonist ZLN005 (10 μg/mL) for 24 h and then challenged with 150 μg/mL LPS for 12 h. Fourth, cells were initially treated with the PGC1α inhibitor SR-18292 (100 μ M) for 6 h followed by a 6-h culture with or without 50 MOI Ad-SIRT3 and a challenge with 150 μg/mL LPS for 12 h. Data were analyzed using one-way ANOVA with subsequent Bonferroni correction. Linear and quadratic contrasts were used to determine dose-responses to LPS. There were linear and quadratic effects of LPS dosage on cell viability. Incubation with 150 and 200 μg/mL LPS for 12 h decreased cell viability to 78.6 and 34.9%, respectively. Compared with controls, expression of IL1B , IL6 , and TNFA was upregulated by 5.2-, 5.9-, and 2.7-fold with 150 μg/mL LPS; concentrations of IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) in cell medium also increased. Compared with the LPS group, LPS+APDC increased cell viability and reversed the upregulation of IL1B , IL6 , and TNFA expression. However, mRNA and protein abundance of SIRT3 decreased linearly with increasing LPS dose. Ad-SIRT3 infection (50 MOI) reduced IL1B , IL6 , and TNFA expression and also their concentrations in cell medium, and decreased pNFκB P65/NFκB P65 ratio and nuclear abundance of NFκB P65. The PGC1α agonist increased SIRT3 expression, whereas it decreased cytokine expression, pNFκB P65/NFκB P65 ratio, and prevented NFκB P65 nuclear translocation. Contrary to the agonist, the PGC1α inhibitor had opposite effects, and elevated the concentrations of IL-1β, IL-6, and TNF-α in cell medium. Overall, data suggested that SIRT3 activity could attenuate LPS-induced inflammatory responses in mammary cells via alterations in the PGC1α-NFκB pathway. As such, there may be potential benefits for targeting SIRT3 in vivo to help prevent or alleviate negative effects of mastitis. [ABSTRACT FROM AUTHOR]

Activation of Kupffer cells (KCs) by gut-derived lipopolysaccharide (LPS) instigates nuclear factor-κB (NF-κB)-mediated inflammatory responses in alcohol-associated liver diseases (ALD). Here, we utilized a novel optogenetically engineered exosome technology called 'exosomes for protein loading via optically reversible protein–protein interactions (EXPLOR)' to efficiently deliver the super-repressor IκB-loaded exosomes (Exo-srIκB) to the liver and examined its therapeutic potential in acute-on-chronic alcohol-associated liver injury. We detected enhanced uptake of DiI-labeled Exo-srIκB by LPS-treated inflammatory KCs, which suppressed LPS-induced inflammatory gene expression levels. In animal experiments, a single intravenous injection of Exo-srIκB prior to alcohol binge drinking significantly attenuated alcohol-associated hepatic steatosis and infiltration of neutrophils and macrophages but not a liver injury. Notably, three consecutive days of Exo-srIκB injection remarkably reduced alcohol-associated liver injury, steatosis, apoptosis of hepatocytes, fibrosis-related gene expression levels in hepatic stellate cells, infiltration of neutrophils and macrophages, and inflammatory gene expression levels in hepatocytes and KCs. In particular, the above effects occurred with inhibition of nuclear translocation of NF-κB in liver tissues, and these beneficial effects of Exo-srIκB on ALD were shown regardless of doses. Our results suggest an exosome-based modulation of NF-κB activity in KCs by Exo-srIκB as a novel and efficient therapeutic approach in ALD. [ABSTRACT FROM AUTHOR]

Neuroinflammation is involved in a wide range of brain disorders, thus there is great interest in identifying novel anti-inflammatory agents to include in therapeutic strategies. Our previous in vitro studies revealed that beta-funaltrexamine (β-FNA), a well-characterized selective mu-opioid receptor (MOR) antagonist, inhibits inflammatory signaling in human astroglial cells, albeit through an apparent MOR-independent mechanism. We also previously determined that lipopolysaccharide (LPS)-induced sickness behavior and neuroinflammation in mice are prevented by pretreatment with β-FNA. Herein we investigated the temporal importance of β-FNA treatment in this pre-clinical model of LPS-induced neuroinflammation. Adult, male C57BL/6J mice were administered an i.p. injection of LPS followed by treatment (i.p. injection) with β-FNA immediately or 4 h post-LPS. Sickness behavior was assessed using an open-field test, followed by assessment of inflammatory signaling in the brain, spleen, and plasma. Levels of inflammatory chemokines/cytokines (interferon γ-induced protein, CXCL10; monocyte chemotactic protein 1, CCL2; and interleukin-6, IL-6) in tissues were measured using an enzyme-linked immunosorbent assay and nuclear factor-kappa B (NFκB), p38 mitogen activated kinase (p38 MAPK), and glial fibrillary acidic protein (GFAP) expression were measured by western blot. LPS-induced sickness behavior and chemokine expression were inhibited more effectively when β-FNA treatment occurred immediately after LPS administration, as opposed to 4 h post-LPS; and β-FNA-mediated effects were time-dependent as evidenced by inhibition at 24 h, but not at 8 h. The inhibitory effects of β-FNA on chemokine expression were more evident in the brain versus the spleen or plasma. LPS-induced NFκB-p65 and p38 MAPK expression in the brain and spleen were inhibited at 8 and 24 h post-LPS. These findings extend our understanding of the anti-inflammatory effects of β-FNA and warrant further investigation into its therapeutic potential. [ABSTRACT FROM AUTHOR]

Objective: To analysis the correlation between nuclear factor-κB (NF-κB) p65 and programmed cell death-1(PD-1), programmed death ligand-1 (PD-L1) in non-small cell lung cancer (NSCLC) tissues and its prognostic value. Methods: 80 NSCLS cases clinical datas and pathological specimens from July 2014 to June 2017 were collected as research objects, another 30 normal lung tissues were collected as control group, the expression of NF-κB p65, PD-1 and PD-L1 in tissues were detected by immunohistochemistry method, the influencing factors of PD-1 and PD-L1 and correlation were analysed, the influencing factors of prognosis were analysed by Logistic regression analysis, the correlation between p65, PD-1, PD-L1 and prognosis was analysed. Results: The positive rates of p65, PD-1 and PD-L1 in NSCLC tissues were higher than in normal control tissues (P＜0.05). TNM stage, differentiation extent, lymph node metastasis, expression of p65 were influencing factors of expressions of PD-1 and PD-L1 (P＜0.05). The correlation analysis showed that p65 was positively correlated with PD-1 and PD-L1 (P＜0.05). The prognosis of NSCLC patients were independently associated with TNM stage, differentiation extent, lymph node metastasis, expression of p65, PD-1 and PD-L1 (P＜0.05). The overall survival (OS) of negative expression patients of p65, PD-1 and PD-L1 was longer than positive expression patients (P＜0.05). Conclusion: The expression of NF-κB p65 was closely correlated with PD-1 and PD-L1, and the expression of p65, PD-1 and PD-L1 had superior predictive value for prognosis of NSCLC patients. [ABSTRACT FROM AUTHOR]

The cytokine oncostatin M (OSM) is regarded as a critical mediator in various inflammatory responses. While the gaseous signaling molecule hydrogen sulfide (H2S) plays a role in a variety of pathophysiological conditions, such as hypertension, inflammatory pain, osteoarthritis, ischemic stroke, oxidative stress, retinal degeneration, and inflammatory responses, the underlying mechanism of H2S action on OSM expression in neutrophils needs to be clarified. In this work, we studied how H2S reduces OSM expression in neutrophil-like differentiated (d)HL-60 cells. To evaluate the effects of H2S, sodium hydrosulfide (NaHS, a donor that produces H2S), ELISA, real-time PCR (qPCR), immunoblotting, and immunofluorescence staining were utilized. Although exposure to granulocyte–macrophage colony-stimulating factor (GM-CSF) resulted in upregulated levels of production and mRNA expression of OSM, these upregulated levels were reduced by pretreatment with NaHS in dHL-60 cells. Similarly, the same pretreatment lowered phosphorylated levels of phosphatidylinositol 3-kinase, Akt, and nuclear factor-kB that had been elevated by stimulation with GM-CSF. Overall, our results indicated that H2S could be a therapeutic agent for inflammatory disorders via suppression of OSM. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of 6% hydroxyethyl starch（HES） 130/0.4 on the inflammatory mediators and its signal pathway micro RNA（miR）-146a/Toll-like receptor 4（TLR4）/nuclear factor-κB（NF-κB） in traumatic rabbits.Methods A total of 16 New Zealand male rabbits were selected in April 2022.After anesthesia, the hind limbs of the rabbits were beaten with blunt objects to cause hind limb fractures, and then the fractures were treated with open internal fixation.The skin was prepared and disinfected, the lateral incision of the fracture site was exposed, and the femur fracture site was exposed, after that a suitable domestic four-hole steel plate was placed across the fracture line.A soft tissue defect model was made in the muscle tissue corresponding to the fracture of about 2 cm×2 cm, and at the same time, the muscle tissue of the blood fracture was taken for backup examination（T0）.According to random number table method, the rabbits were divided into the sodium lactate Ringer（LR） group and the HES group, with eight rabbits in each group.The LR group was injected with 10 mL/kg LR,while the HES group was injected 10 mL/kg HES once a day for five days.After the infusion rabbits were anesthetized, the muscle tissue of the injured site was taken for backup examination（T1）.The enzyme-linked immunosorbent assay（ELISA） was used to determine tumor necrosis factor-α（TNF-α）,interleukin-1（IL-1） and IL-6 in blood of rabbits at T0 and T1,and western blotting was used to determine NF-κB P65,TLR4,IL-1 receptor-associated kinase 1（IRAK1）,and TNF receptor-associated factor 6（TRAF6） protein levels in the muscle tissue at T0 and T1.The reverse transcription-polymerase chain reaction was used to determine miR-146a in the rabbit muscle tissue at T0 and T1.Results Compared with T0,the levels of TNF-α,IL-1,IL-6 in blood, and the levels of miR-146a, NF-κB P65,TLR4,IRAK1,and TRAF6 protein in muscle of the two groups were significantly higher at T1,and the differences were statistically significant（P<0.05）.Compared with the LR group, the levels of TNF-α,IL-1 and IL-6 in blood, NF-κB P65,TLR4,IRAK1 and TRAF6 protein in muscle of the rabbits in the HES group were significantly lower at T1,while the level of miR-146a in muscle was significantly higher, with statistical significance differences（P<0.05）.Conclusion HES may upregulate miR-146a, negatively feedback to regulate IRAK1 and TRAF6 in the TLR4 signal pathway, and inhibit the production of inflammatory mediators, thus reducing the inflammatory response of traumatic rabbits. [ABSTRACT FROM AUTHOR]

Background: Inflammation is present in neurological and peripheral disorders. Thus, targeting inflammation has emerged as a viable option for treating these disorders. Previous work indicated pretreatment with beta-funaltrexamine (β-FNA), a selective mu-opioid receptor (MOR) antagonist, inhibited inflammatory signaling in vitro in human astroglial cells, as well as lipopolysaccharide (LPS)-induced neuroinflammation and sickness-like-behavior in mice. This study explores the protective effects of β-FNA when treatment occurs 10 h after LPS administration and is the first-ever investigation of the sex-dependent effects of β-FNA on LPS-induced inflammation in the brain and peripheral tissues, including the intestines. Results: Male and female C57BL/6J mice were administered LPS followed by treatment with β-FNA-immediately or 10 h post-LPS. Sickness- and anxiety-like behavior were assessed using an open-field test and an elevated-plus-maze test, followed by the collection of whole brain, hippocampus, prefrontal cortex, cerebellum/brain stem, plasma, spleen, liver, large intestine (colon), proximal small intestine, and distal small intestine. Levels of inflammatory chemokines/cytokines (interferon γ-induced-protein, IP-10 (CXCL10); monocyte-chemotactic-protein 1, MCP-1 (CCL2); interleukin-6, IL-6; interleukin-1β, IL-1β; and tumor necrosis factor-alpha, TNF-α) in tissues were measured using an enzyme-linked immunosorbent assay. Western blot analysis was used to assess nuclear factor-kappa B (NF-κB) expression. There were sex-dependent differences in LPS-induced inflammation across brain regions and peripheral tissues. Overall, LPS-induced CXCL10, CCL2, TNF-α, and NF-κB were most effectively downregulated by β-FNA; and β-FNA effects differed across brain regions, peripheral tissues, timing of the dose, and in some instances, in a sex-dependent manner. β-FNA reduced LPS-induced anxiety-like behavior most effectively in female mice. Conclusion: These findings provide novel insights into the sex-dependent anti-inflammatory effects of β-FNA and advance this agent as a potential therapeutic option for reducing both neuroinflammation an intestinal inflammation. [ABSTRACT FROM AUTHOR]

The epithelium-derived cytokines interleukin (IL)-25, IL-33, and thymic stromal lymphopoietin (TSLP) are important mediators that initiate innate type 2 immune responses in asthma. Leukotriene receptor antagonists (LTRAs) are commonly used to prevent asthma exacerbations. However, the effects of LTRAs on epithelium-derived cytokines expression in airway epithelial cells are unclear. This study aimed to investigate the effects of LTRAs on the expression of epithelium-derived cytokines in human airway epithelial cells and to explore possible underlying intracellular processes, including epigenetic regulation. A549 or HBE cells in air-liquid interface conditions were pretreated with different concentrations of LTRAs. The expression of epithelium-derived cytokines and intracellular signaling were investigated by real-time PCR, enzyme-linked immunosorbent assay, and Western blot. In addition, epigenetic regulation was investigated using chromatin immunoprecipitation analysis. The expression of IL-25, IL-33, and TSLP was increased under LTRAs treatment and suppressed by inhaled corticosteroid cotreatment. Montelukast-induced IL-25, IL-33, and TSLP expression were mediated by the mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways and regulated by histone H3 acetylation and H3K36 and H3K79 trimethylation. LTRAs alone might increase inflammation and exacerbate asthma by inducing the production of IL-25, IL-33, and TSLP; therefore, LTRA monotherapy may not be an appropriate therapeutic option for asthma. [ABSTRACT FROM AUTHOR]

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Background: SET domain containing 6 (SETD6) has been shown to be upregulated in multiple human cancers and can promote malignant cell survival. However, expression and function of SETD6 in lung adenocarcinoma (LUAD) remains unaddressed. This study aimed to demonstrate the expression pattern, biological roles and potential mechanisms by which SETD6 dysregulation is associated with LUAD. Methods: The expression level of SETD6 was evaluated in LUAD clinical specimens and its correlation with clinical parameters were analyzed. In vitro, gain-of-function and loss-of-function experiments were performed to evaluate the effects of SETD6 on cell proliferation, apoptosis, migration, and colony formation of LUAD cell line A549. Western-blot was performed to investigate the involvement of nuclear factor-κB (NF-κB) and nuclear factor erythroid 2–related factor 2 (Nrf2) pathways as downstream signaling of SETD6 in LUAD cells. Results: Compared with non-tumorous tissues, SETD6 was overexpressed in tumor tissues, and its overexpression significantly correlates with higher rates of regional lymph node metastasis and poor prognosis in patients with LUAD. In A549 cell line, SETD6 overexpression could promote cell proliferation, migration, colony formation and inhibit cell apoptosis, whereas SETD6 knockdown caused the opposite effects. Furthermore, we demonstrated that the mechanisms underlying the effect of SETD6 on LUAD biological behaviors may be through its interaction with NF-κB and Nrf2 signaling pathways. Conclusions: SETD6, which is highly expressed in LUAD tumor tissues, plays an important role in promoting the malignant behaviors of LUAD via likely the NF-κB and Nrf2 signaling pathways. [ABSTRACT FROM AUTHOR]

Objective: To investigate the regulatory effects of Survival protein (Survivin) on the expression of trans⁃ forming growth factor β-activated kinase 1 (Tak1) and nuclear factor κB (NF-κB), cell cycle, and apoptosis of esophageal cancer cells. Methods: Human esophageal carcinoma cells Eca109 were cultured in vitro and divided into the blank con⁃ trol group, Survivin-siRNA group, Survivin-siRNA empty vector group, Survivin overexpression group, Survivin overex⁃ pression empty vector group, and YM155 group. Cells in the Survivin-siRNA and Survivin-siRNA empty vector groups were transfected with Survivin siRNA vector or empty vector of Survivin siRNA, respectively. Cells in the Survivin overex⁃ pression group and the Survivin overexpression empty vector group were transfected with the overexpression vector of Sur⁃ vivin or the overexpression empty vector of Survivin for 6 h, respectively, and then they were cultured in a fresh medium for 24 h to collect the cells. Cells in the blank control group were not transfected, and cells in the YM155 group were added with Survivin inhibitor YM155. After that, we collected the cells in each transfection group simultaneously. The expres⁃ sion levels of Survivin, phosphorylated Tak1 (p-Tak1), and NF-κB p65 were detected by Western blotting, and the cell cycle distribution and apoptosis rate were detected by flow cytometry. Results: Compared with the blank control group，the relative expression of Survivin significantly increased in the Survivin overexpression group but significantly decreased in the Survivin-siRNA group and YM155 group (all P<0. 05) . Compared with the blank control group, the expression lev⁃ els of p-TAK1 and NF-κB protein in each transfection group and YM155 group had no significant changes (all P>0. 05) . Compared with the blank control group, the proportion of cells in G2 phase significantly increased (P<0. 05), the propor⁃ tion of cells in S phase significantly decreased (P<0. 05), and the proportion of cells in the G1 phase had no significant change in the YM155 group (all P>0. 05) . Compared with the blank control group, there were no significant changes in the proportions of cells in the G1, G2, and S phases in each transfection group (all P>0. 05) . Compared with the blank con⁃ trol group, the early apoptosis rate and total apoptosis rate of the Survivin-siRNA group increased (both P<0. 05), but the late apoptosis rate had no significant change (P>0. 05) . Compared with the blank control group, there were no significant changes in the early apoptosis rate, late apoptosis rate, or total apoptosis rate in the other transfection groups and the YM155 group (all P>0. 05) . Conclusion: Survivin expression is involved in regulating cell cycle and apoptosis in esoph⁃ ageal cancer cells, but its effect is not implemented by regulating Tak1 and NF-κB. [ABSTRACT FROM AUTHOR]

Objective： To explore the improvement effects of grape seed proanthocyanidin extract (GSPE) on the periodontal inflammation in the diabetic periodontitis rats, and to clarify its mechanism. Methods：Forty male SD rats were randomly divided into control group, diabetic periodontitis model group (model group), low dose (100 mg·kg－1) of GSPE group and high dose (200 mg·kg－1) of GSPE group； there were 10 rats in each group. Except for control group, the rats in other groups were used to establish the diabetic periodontitis models by periodontal ligation combined with intraperitoneal injection of streptozotocin (STZ). After the successful establishment of the models, the rats in low and high doses of GSPE groups were given 100 and 200 mg·kg－1 GSPE, respectively, while the rats in control group and model group were given the same amount of normal saline once a day for 8 weeks. The general situation of the rats and the changes of blood glucose levels were observed, the pathomorphology of periodontal tissue of the rats in various groups were observed by HE staining, the alveolar bone absorption of the rats in various groups was measured, the activities of superoxide dismutase (SOD) and the catalase (CAT) and the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in periodontal tissue of the rats in various groups were detected by related kits. Enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β) and interleukin-6 (IL-6) in serum of the rats in various groups. The expression levels of Toll-like receptor 4 (TLR4) and nuclear factor- κB (NF- κB) proteins in periodontal tissue of the rats in various groups were detected by Western blotting method. Results：Compared with control group, the rats in model group showed obvious symptoms of“ three more and one less”；the level of blood glucose of the rats was increased obviously (P< 0. 05), the inflammatory cells were infiltrated in periodontal connective tissue, periodontal tissue was damaged, and the alveolar bone absorption amount was increased (P<0. 05)；the activities of SOD and CAT in periodontal tissue were significantly decreased (P<0. 05), the levels of ROS and MDA were significantly increased (P<0. 05), the levels of TNF- α, IL-1β and IL-6 in serum were significantly increased (P<0. 05), and the expression levels of TLR4 and NF- κ B proteins were significantly increased (P<0. 05). Compared with model group, the general condition of the rats in low and high doses of GSPE groups was obviously improved, the levels of blood glucose were reduced (P<0. 05), periodontal tissue damage was obviously reduced, the alveolar bone absorption amounts were reduced (P<0. 05), the activities of SOD and CAT in periodontal tissue were significantly increased (P<0. 05), the levels of ROS and MDA were significantly decreased (P<0. 05), the levels of TNF- α, IL-1β and IL-6 in serum were significantly decreased (P<0. 05), and the expression levels of TLR4 and NF-κB proteins were significantly decreased (P<0. 05). Compared with low dose of GSPE group, the activities of SOD and CAT in periodontal tissue of the rats in high dose of GSPE group were significantly increased (P<0. 05), the levels of ROS and MDA were significantly decreased (P<0. 05), the levels of TNF-α, IL-1β and IL-6 in serum were significantly decreased (P<0. 05), and the expression levels of TLR4 and NF- κB proteins were significantly decreased (P<0. 05). Conclusion：GSPE can improve the periodontal inflammation in the diabetic periodontitis rats, and its mechanism may be related to regulating the TLR4 and NF- κB signaling pathway in periodontal tissue. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of nootkatone (NKT) on chronic unpredictable mild stress (CUMS)-induced depressive-like behaviors and the mechanism underlying NKT improving the depressive-like behaviors. Methods: The CUMS-induced depression model was established in mice. Fifty mice were randomized into 5 groups (n=10) in accordance with a random number table: control group, CUMS group, CUMS + NKT (6 mg/kg) group, CUMS + NKT (12 mg/kg) group, and CUMS + ketamine group. From the 22th day, NKT (6 or 12 mg/kg) or ketamine (0.5 mg/kg) was given with intragastric administration every day for 21 days. Behavioral tests including forced swimming test (FST), tail suspension test (TST), sucrose preference test (SPT) and open-field test (OFT) were carried out. The mRNA and protein expressions of interleukin (IL)-1β, IL-18, IL-6, and tumor necrosis factor (TNF)-α in hippocampus were assessed using quantitative realtime polymerase chain reaction (PCR), Western blot analysis, and enzyme linked immunosorbent assay. The nuclear factor-κB (NF-κB)/NOD-like receptor 3 (NLRP3) inflammasome pathway was analyzed using Western blot and immunofluorescence analysis. Results: NKT treatment improved CUMS-induced depressive-like behaviors in mice (P<0.05 or P<0.01). NKT significantly decreased the mRNA and protein levels of IL-1β, IL-18, IL-6, and TNF-α in hippocampus of CUMS mice (P<0.05 or P<0.01). Furthermore, NKT repressed CUMS-induced activation of NF-κB signaling and NLRP3 inflammasome (P<0.01). More important, Nigericin, a NLRP3 activator, destroyed the effect of NKT on repressing neuroinflammation and improving depressive-like behaviors (P<0.05 or P<0.01). Conclusion: NKT ameliorates the depressive-like symptoms, in part by repressing NF-κB/NLRP3-mediated neuroinflammation. [ABSTRACT FROM AUTHOR]

Background: Intestinal barrier integrity in the pathogenesis of sepsis is critical. Despite an abundance of evidence, the molecular mechanism of the intestinal barrier in sepsis pathology remains unclear. Here, we report a protective role of polo-like kinase 1 (PLK1) in intestinal barrier integrity during sepsis. Methods: Mice with PLK1 overexpression (CAG-PLK1 mice) or PLK1 inhibition (BI2536-treated mice) underwent caecal ligation and puncture (CLP) to establish a sepsis model. The intestinal barrier function, apoptosis in the intestinal epithelium, mitochondrial function and NF-κB signalling activity were evaluated. To suppress the activation of NF-κB signalling, the NF-κB inhibitor PDTC, was administered. The Caco-2 cell line was chosen to establish an intestinal epithelial injury model in vitro. Results: Sepsis destroyed intestinal barrier function, induced excessive apoptosis in the intestinal epithelium, and disrupted the balance of mitochondrial dynamics in wild-type mice. PLK1 overexpression alleviated sepsis-induced damage to the intestinal epithelium by inhibiting the activation of NF-κB signalling. PLK1 colocalized and interacted with TANK in Caco-2 cells. Transfecting Caco-2 cells with TANK-SiRNA suppressed NF-κB signalling and ameliorated mitochondrial dysfunction, apoptosis and the high permeability of cells induced by lipopolysaccharide (LPS). Furthermore, TANK overexpression impaired the protective effect of PLK1 on LPS-induced injuries in Caco-2 cells. Conclusion: Our findings reveal that the PLK1/TANK/NF-κB axis plays a crucial role in sepsis-induced intestinal barrier dysfunction by regulating mitochondrial dynamics and apoptosis in the intestinal epithelium and might be a potential therapeutic target in the clinic. [ABSTRACT FROM AUTHOR]

Mastitis is a common inflammatory cow mammary infection; that causes significant economic loss in dairy industry. Given the interesting connection between metformin's anti-inflammatory function and mastitis model induced by LTA in pbMECs, our objective was to prove that metformin was beneficial in suppressing proinflammatory response induced by LTA through modulation of mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways and activation of peroxisome proliferator-activated receptor-γ (PPARγ) in pbMECs. The proliferation of cells and mRNA expression were measured using EdU assay and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Immunoblotting and immunofluorescence analysis were conducted to evaluate the expression of target proteins in inflammatory and anti-inflammatory responses to metformin and LTA. Finally, pbMECs were allowed to treat with the PPAR antagonist GW9662, and inflammatory markers were detected in the cells. Our results showed that LTA concentration at 100 µg/mL significantly stimulated the MAPK14, IL-6 and IL-1β mRNA expressions compared to the control cells (P < 0.05) in dose-dependent tests for LTA. Metformin suppressed the phosphorylation expressions of MAPK (ERK1/2, p38, and JNK) in LTA-stimulated pbMECs. Metformin also reduced the protein expression of NF-κ B, interleukin-8 (IL-8), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in pbMECs pretreated with LTA. Metformin administration activated PPARγ phosphorylation by up-regulating the expression of PPARγ in LTA-stimulated pbMECs. Treatment with GW9662 resulted in increased IL-6 expression, which was reversed by metformin. These findings collectively indicated that metformin act to attenuate LTA-stimulated inflammatory response in pbMECs by suppressing MAPK and NF-κ B activation via a mechanism partially dependent on PPARγ activation. These results suggested that metformin could function as an anti-inflammatory drug in the treatment of mastitis. [ABSTRACT FROM AUTHOR]

Background: The NF-κB signaling cascade regulates immune response and is often dysregulated in tumor development. UBE2W is a novel type I ubiquitin-conjugating enzyme (E2) whose biological function is still unclear. Aims: This study was designed to investigate whether UBE2W regulates NF-κB signaling pathway and is involved in the progression of experimental colitis. Methods: At the cellular level, the effect of UBE2W on NF-κB transcriptional activity was measured using a dual-luciferase reporter assay. The influence of UBE2W on NF-κB pathway activation and the entry of p65 into the nucleus were determined by Western blot and immunofluorescence analyses, respectively. Moreover, the colitis model was established by administering 2.5% dextran sulfate sodium (DSS)/water to UBE2W overexpression, UBE2W-knockdown and control mice. Body weight, stool consistency, colon length and clinical severity were examined. Expression of pro-inflammatory cytokines and phosphorylation of p65 and IκB in the colon tissue were measured by qRT-PCR and Western blot, respectively. Results: UBE2W inhibited TNFα-induced NF-κB transcription activity, attenuated IκB and p65 phosphorylation, downregulated TNFα and IL-8 expression and blocked the entry of p65 into the nucleus. In the DSS-induced colitis model, UBE2W-knockdown mice had increased weight loss, more serious diarrhea and mucosal injures compared with the control mice. Moreover, phosphorylation of IκB and p65 and the expression of pro-inflammatory mediators such as TNFα, IL-6 were significantly increased in UBE2W knockdown mice. However, these changes were completely reversed in UBE2W overexpression mice. Conclusions: The overexpression of UBE2W ameliorates the severity of DSS-induced colitis, which may be mediated by inhibiting the expression of pro-inflammatory mediators and activation of the NF-κB signaling pathway. These findings provide evidence that UBE2W might have potential therapeutic implications in IBD. [ABSTRACT FROM AUTHOR]

Nuciferine, isolated from Nelumbo nucifera (commonly known as lotus) leaves, has been shown to have beneficial effects, including antioxidant, anti-obesity, anti-diabetic, and anti-inflammatory properties. However, little is known about the mechanism of nuciferine action on the inflammatory response. This study aimed to investigate the anti-inflammatory effects of nuciferine and its underlying molecular mechanisms in lipopolysaccharide (LPS)-stimulated murine macrophages. In this study, nuciferine reduced LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production and mRNA expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Nuciferine also decreased the production of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. Furthermore, nuciferine inhibited the LPS-mediated transcriptional activity of nuclear factor (NF)-κB and activator protein (AP)-1, and the nuclear translocation of NF-κB p65 and activating transcription factor 2 (ATF2), an AP-1 subunit. Nuciferine also decreased the phosphorylation of IκB kinase (IKK), inhibitor of NF-κB (IκB), NF-κB, mitogen-activated protein kinase 3 (MKK3), MKK6, p38 mitogen-activated protein kinase (MAPK), and ATF2. Overall, our findings suggest that nuciferine may exert anti-inflammatory effects in LPS-induced macrophages by inhibiting the NF-κB and p38 MAPK/ATF2 signaling pathways. [ABSTRACT FROM AUTHOR]

Levosimendan, a calcium sensitizer, has an organ protective profile through the inhibition of inflammatory mediators and cytokines in critical conditions, such as heart failure, ischemia-reperfusion injury, and sepsis. The survival effect of levosimendan for acute liver failure has not been examined yet. Male Sprague-Dawley rats were examined in the D-galactosamine hydrochloride and lipopolysaccharide (GalN/LPS) model. Levosimendan was injected intraperitoneally before GalN/LPS treatment. Survival was monitored for 7 days. For biochemical analyses, liver and blood samples were collected from the rats at 1 and 8 h after GaIN/LPS treatment. The pretreatment of levosimendan at 4 mg/kg significantly increased survival in GalN/LPS rats. In the liver specimen, levosimendan significantly inhibited the activation of nuclear factor-κB (NF-κB) at 1 h, and significantly decreased the mRNA expression of inflammatory mediators, including inducible nitric oxide synthase and tumor necrosis factor-α (TNF-α), at 8 h. In serum, levosimendan decreased the levels of nitrite, a metabolite of nitric oxide, and TNF-α protein, as well as aspartate aminotransferase and alanine aminotransferase. These results indicated that Levosimendan ameliorated liver dysfunction and survival in acute liver failure model rats through the suppression of NF-κB activation. [ABSTRACT FROM AUTHOR]