SARS-CoV-2 Spike Glycoprotein S1 Induces Neuroinflammation in BV-2 Microglia

AbstractIn addition to respiratory complications produced by SARS‐CoV‐2, accumulating evidence suggests that some neurological symptoms are associated with the disease caused by this coronavirus. In this study, we investigated the effects of the SARS‐CoV‐2 spike protein S1 stimulation on neuroinflammation in BV-2 microglia. Analyses of culture supernatants revealed an increase in the production of TNF-α, IL-6, IL-1β and iNOS/NO. S1 also increased protein levels of phospho-p65 and phospho-IκBα, as well as enhanced DNA binding and transcriptional activity of NF-κB. These effects of the protein were blocked in the presence of BAY11-7082 (1 µM). Exposure of S1 to BV-2 microglia also increased the protein levels of NLRP3 inflammasome and enhanced caspase-1 activity. Increased protein levels of p38 MAPK was observed in BV-2 microglia stimulated with the spike protein S1 (100 ng/ml), an action that was reduced in the presence of SKF 86,002 (1 µM). Results of immunofluorescence microscopy showed an increase in TLR4 protein expression in S1-stimulated BV-2 microglia. Furthermore, pharmacological inhibition with TAK 242 (1 µM) and transfection with TLR4 small interfering RNA resulted in significant reduction in TNF-α and IL-6 production in S1-stimulated BV-2 microglia. These results have provided the first evidence demonstrating S1-induced neuroinflammation in BV-2 microglia. We propose that induction of neuroinflammation by this protein in the microglia is mediated through activation of NF-κB and p38 MAPK, possibly as a result of TLR4 activation. These results contribute to our understanding of some of the mechanisms involved in CNS pathologies of SARS-CoV-2.

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Skullcapflavone II Suppresses TNF-α/IFN-γ-Induced TARC, MDC, and CTSS Production in HaCaT Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22126428 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6428

Author(s):

Hanon Lee ◽

Dong Hun Lee ◽

Jang-Hee Oh ◽

Jin Ho Chung

Keyword(s):

P38 Mapk ◽

Therapeutic Potential ◽

Inflammatory Diseases ◽

Mapk Signaling ◽

Hacat Cells ◽

Protein Levels ◽

Tnf Α ◽

Ifn Γ ◽

Mapk Signaling Pathways ◽

Pro Inflammatory Cytokines

Skullcapflavone II (SFII), a flavonoid derived from Scutellaria baicalensis, has been reported to have anti-inflammatory properties. However, its therapeutic potential for skin inflammatory diseases and its mechanism are unknown. Therefore, this study aimed to investigate the effect of SFII on TNF-α/IFN-γ-induced atopic dermatitis (AD)-associated cytokines, such as thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Co-stimulation with TNF-α/IFN-γ in HaCaT cells is a well-established model for induction of pro-inflammatory cytokines. We treated cells with SFII prior to TNF-α/IFN-γ-stimulation and confirmed that it significantly inhibited TARC and MDC expression at the mRNA and protein levels. Additionally, SFII also inhibited the expression of cathepsin S (CTSS), which is associated with itching in patients with AD. Using specific inhibitors, we demonstrated that STAT1, NF-κB, and p38 MAPK mediate TNF-α/IFN-γ-induced TARC and MDC, as well as CTSS expression. Finally, we confirmed that SFII significantly suppressed TNF-α/IFN-γ-induced phosphorylation of STAT1, NF-κB, and p38 MAPK. Taken together, our study indicates that SFII inhibits TNF-α/IFN-γ-induced TARC, MDC, and CTSS expression by regulating STAT1, NF-κB, and p38 MAPK signaling pathways.

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Multiple Myeloma Cells Survival and Proliferation Rely on High Levels and Activity of the Serine-Threonine Kinase CK2.

Blood ◽

10.1182/blood.v104.11.643.643 ◽

2004 ◽

Vol 104 (11) ◽

pp. 643-643 ◽

Cited By ~ 1

Author(s):

Francesco A. Piazza ◽

Maria Ruzzene ◽

Giovanni Di Maira ◽

Enrico Brunetta ◽

Luca Bonanni ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Progression ◽

Transcriptional Activity ◽

Kinase Activity ◽

Serine Threonine Kinase ◽

Cycle Progression ◽

Threonine Kinase ◽

Protein Levels ◽

Tnf Α ◽

Ck2 Inhibitors

Abstract Survival and proliferation of Multiple Myeloma plasma cells (MMPCs) depend on the activation of signaling pathways through the interaction with the surrounding bone marrow microenvironment. CK2 is a ubiquitous cellular serine-threonine kinase, whose involvement in oncogenic transformation, apoptosis and cell cycle progression has recently become matter of intense research. Due to its connection with signaling molecules pivotal for plasma cell (PCs) survival, such as those implicated in the TNF-α/NF-κB, IGF1/PI3K/AKT and Wnt/β-catenin pathways, CK2 is likely to play a central role in MM biology. We investigated CK2 function in MMPCs survival and cell cycle progression, in the modulation of the sensitivity to chemotherapeutics and in the regulation of the I-κB/NF-κB dependent pathway. We first analysed the CK2 protein levels and specific kinase activity in MMPCs. Different cell lines and highly purified CD138+ PCs from 5 patients were used. We observed higher protein levels of the CK2 catalytic subunit αin the neoplastic MMPCs than in controls (resting peripheral blood and splenic B lymphocytes). Moreover, also the total CK2-dependent kinase activity was found significantly increased in MMPCs. We also assessed the levels and pattern of total protein phosphorylation by radioactive phosphate incorporation assay. We found that MMPCs share a similar pattern of phoshorylated proteins. The degree of phosphorylation of some of these proteins was significantly reduced in the presence of specific CK2 inhibitors. Next, using a panel of highly specific CK2 inhibitors, we studied the effects of hampering CK2 function in MMPCs. A dose-dependent cytotoxic effect was observed after the treatment with such compounds that was associated with the activation of both the extrinsic and intrinsic caspase-dependent pathways, the release from mitochondria of cytochrome c and smac/diablo and cell cycle arrest in G2-M. A possible role for CK2 inhibition in sensitising MMPCs to melphalan-induced apoptosis was also investigated. Indeed, CK2 blockade lowered the threshold of sensitivity of MMPCs to the cytotoxic effect of melphalan. We then looked at the consequences of CK2 blockade on the NF-κB dependent signaling cascade. Basal and TNF-α-dependent I-κB-αdegradation, as well as NF-κB transcriptional activity upon TNF-αstimulation, were partially impaired by CK2 blockade in MMPCs. Finally, we detected association between the endogenous αcatalytic subunit of CK2 and the NF-κB p50/p105 member by confocal microscopy and co-immunoprecipitation. Altogether, our data suggest a pivotal role for CK2 in controlling survival, proliferation and sensitivity to chemotherapeutics of MMPCs and implicate this kinase in the regulation of the NF-κB pathway in MM through the modulation of I-κB protein levels and NF-κB transcriptional activity. This latter effect is possibly exerted through physical association of CK2 with NF-κB transcription factors. Our findings also suggest that CK2 inhibition could be exploited as a novel therapeutic approach for MM.

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Deoxynivalenol Induces Inflammation in IPEC-J2 Cells by Activating P38 Mapk And Erk1/2

Toxins ◽

10.3390/toxins12030180 ◽

2020 ◽

Vol 12 (3) ◽

pp. 180 ◽

Cited By ~ 2

Author(s):

Hua Zhang ◽

Xiwen Deng ◽

Chuang Zhou ◽

Wenda Wu ◽

Haibin Zhang

Keyword(s):

P38 Mapk ◽

Inhibitory Effect ◽

Epithelial Cell Line ◽

P38 Inhibitor ◽

Protein Levels ◽

Extracellular Signal ◽

Tnf Α ◽

Proinflammatory Gene ◽

Upregulated Genes ◽

Proinflammatory Gene Expression

Fusarium-derived mycotoxin deoxynivalenol (DON) usually induces diarrhea, vomiting and gastrointestinal inflammation. We studied the cytotoxic effect of DON on porcine small intestinal epithelium using the intestinal porcine epithelial cell line IPEC-J2. We screened out differentially expressed genes (DEGs) using RNA-seq and identified 320 upregulated genes and 160 downregulated genes. The enrichment pathways of these DEGs focused on immune-related pathways. DON induced proinflammatory gene expression, including cytokines, chemokines and other inflammation-related genes. DON increased IL1A, IL6 and TNF-α release and DON activated the phosphorylation of extracellular signal-regulated kinase-1 and-2 (ERK1/2), JUN N-terminal kinase (JNK) and p38 MAPK. A p38 inhibitor attenuated DON-induced IL6, TNF-α, CXCL2, CXCL8, IL12A, IL1A, CCL20, CCL4 and IL15 production, while an ERK1/2 inhibitor had only a small inhibitory effect on IL15 and IL6. An inhibitor of p38 MAPK decreased the release of IL1A, IL6 and TNF-α and an inhibitor of ERK1/2 partly attenuated protein levels of IL6. These data demonstrate that DON induces proinflammatory factor production in IPEC-J2 cells by activating p38 and ERK1/2.

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Mechanism of estrogen-mediated intestinal protection following trauma-hemorrhage: p38 MAPK-dependent upregulation of HO-1

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00112.2008 ◽

2008 ◽

Vol 294 (6) ◽

pp. R1825-R1831 ◽

Cited By ~ 35

Author(s):

Jun-Te Hsu ◽

Wen-Hong Kan ◽

Chi-Hsun Hsieh ◽

Mashkoor A. Choudhry ◽

Martin G. Schwacha ◽

...

Keyword(s):

P38 Mapk ◽

Critical Role ◽

Cell Types ◽

Intestinal Injury ◽

Male Rats ◽

Protein Levels ◽

Mapk Activity ◽

Inflammatory Protein ◽

Tnf Α ◽

Sb 203580

p38 MAPK has been reported to regulate the inflammatory response in various cell types via extracellular stimuli. p38 MAPK activation also results in the induction of heme oxygenase (HO)-1, which exerts potent anti-inflammatory effects. Although studies have shown that 17β-estradiol (E2) prevented organ dysfunction following trauma-hemorrhage, it remains unknown whether p38 MAPK/HO-1 plays any role in E2-mediated attenuation of intestinal injury under those conditions. To study this, male rats underwent trauma-hemorrhage (mean blood pressure ∼40 mmHg for 90 min) followed by fluid resuscitation. At the onset of resuscitation, rats were treated with vehicle, E2 (1 mg/kg body wt), the p38 MAPK inhibitor SB-203580 (2 mg/kg body wt) or E2 plus SB-203580. Two hours thereafter, intestinal myeloperoxidase (MPO) activity and lactate, TNF-α, IL-6, ICAM-1, cytokine-induced neutrophil chemoattractant (CINC)-1, and macrophage inflammatory protein (MIP)-2 levels were measured. Intestinal p38 MAPK and HO-1 protein levels were also determined. Trauma-hemorrhage led to an increase in intestinal MPO activity and lactate, TNF-α, IL-6, ICAM-1, CINC-1, and MIP-2 levels. This was accompanied with a decrease in intestinal p38 MAPK activity and increase in HO-1 expression. Administration of E2 normalized all the above parameters except HO-1, which was further increased following trauma-hemorrhage. Administration of SB-203580 with E2 abolished the E2-mediated restoration of the above parameters as well as the increase in intestinal HO-1 expression following trauma-hemorrhage. These results suggest that the p38 MAPK/HO-1 pathway plays a critical role in mediating the salutary effects of E2 on shock-induced intestinal injury.

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Proinflammatory responses of human airway cells to ricin involve stress-activated protein kinases and NF-κB

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00207.2007 ◽

2007 ◽

Vol 293 (6) ◽

pp. L1385-L1394 ◽

Cited By ~ 35

Author(s):

John Wong ◽

Veselina Korcheva ◽

David B. Jacoby ◽

Bruce E. Magun

Keyword(s):

P38 Mapk ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Nucleosome Assembly ◽

Human Airway ◽

Tnf Α ◽

Proinflammatory Responses ◽

Pulmonary Damage ◽

Interfering Rna ◽

Airway Cells

Ricin is a potential bioweapon because of its toxicity, availability, and ease of production. When delivered to the lungs, ricin causes severe pulmonary damage with symptoms that are similar to those observed in acute lung injury and adult respiratory distress syndrome. The airway epithelium plays an important role in the pathogenesis of many lung diseases, but its role in ricin intoxication has not been elucidated. Exposure of cultured primary human airway epithelial cells to ricin resulted in the activation of SAPKs and NF-κB and in the increased expression of multiple proinflammatory molecules. Among the genes upregulated by ricin and identified by microarray analysis were those associated with transcription, nucleosome assembly, inflammation, and response to stress. Sequence analysis of the promoters of these genes identified NF-κB as one of the transcription factors whose binding sites were overrepresented. Although airway cells secrete TNF-α in response to ricin, blocking TNF-α did not prevent ricin-induced activation of NF-κB. Decreased levels of IκB-α in airway cells exposed to ricin suggest that translational suppression may be responsible for the activation of NF-κB. Inhibition of p38 MAPK by a chemical inhibitor or NF-κB by short interfering RNA resulted in a marked reduction in the expression of proinflammatory genes, demonstrating the importance of these two pathways in ricin intoxication. Therefore, the p38 MAPK and NF-κB pathways are potential therapeutic targets for reducing the inflammatory consequences of ricin poisoning.

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AAnti-Inflammatory Effects of Plasma Circulating Exosomes Obtained from Normal-Weight and Obese Subjects on Hepatocytes

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530320666200505121426 ◽

2020 ◽

Vol 20 ◽

Author(s):

Reza Afrisham ◽

Sahar Sadegh-Nejadi ◽

Reza Meshkani ◽

Solaleh Emamgholipour ◽

Molood Bagherieh ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Hepg2 Cells ◽

Human Liver ◽

Liver Cells ◽

Normal Weight ◽

Protein Levels ◽

Human Liver Cells ◽

Tnf Α ◽

Anti Inflammatory

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.

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The Impact of Royal Jelly against Hepatic Ischemia/Reperfusion-Induced Hepatocyte Damage in Rats: The Role of Cytoglobin, Nrf-2/HO-1/COX-4, and P38-MAPK/NF-κB-p65/TNF-α Signaling Pathways

Current Molecular Pharmacology ◽

10.2174/1874467213666200514223829 ◽

2020 ◽

Vol 14 (1) ◽

pp. 88-100

Author(s):

Fares E.M. Ali ◽

Heba M. Saad Eldien ◽

Nashwa A.M. Mostafa ◽

Abdulrahman H. Almaeen ◽

Mohamed R.A. Marzouk ◽

...

Keyword(s):

Signaling Pathways ◽

P38 Mapk ◽

Royal Jelly ◽

Ischemia Reperfusion ◽

Histopathological Changes ◽

Down Regulation ◽

Hepatic Ischemia ◽

Tnf Α ◽

Hepatic Ischemia Reperfusion ◽

The Impact

Objective: The present study was conducted to elucidate the underlying molecular mechanism as well as the potential hepatoprotective effects of royal jelly (RJ) against hepatic ischemia/reperfusion (IR) injury. Methods: Rats were assigned into four groups; sham (received vehicle), IR (30 minutes ischemia and 45 minutes reperfusion), sham pretreated with RJ (200 mg/kg P.O.), and IR pretreated with RJ (200 mg/kg P.O.). The experiment has lasted for 28 days. Results: Hepatic IR significantly induced hepatic dysfunctions, as manifested by elevation of serum transaminases, ALP and LDH levels. Moreover, hepatic IR caused a significant up-regulation of P38-MAPK, NF-κB-p65, TNF-α and MDA levels along with marked down-regulation of Nrf-2, HO-1, COX-4, cytoglobin, IκBa, IL-10, GSH, GST and SOD levels. Additionally, marked histopathological changes were observed after hepatic IR injury. On the contrary, pretreatment with RJ significantly improved hepatic functions along with the alleviation of histopathological changes. Moreover, RJ restored oxidant/antioxidant balance as well as hepatic expressions of Nrf-2, HO-1, COX-4, and cytoglobin. Simultaneously, RJ significantly mitigated the inflammatory response by down-regulation of P38-MAPK, NF-κB-p65, TNF-α expression. Conclusion: The present results revealed that RJ has successfully protected the liver against hepatic IR injury through modulation of cytoglobin, Nrf-2/HO-1/COX-4, and P38-MAPK/NF-κB-p65/TNF-α signaling pathways.

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Let-7a-5p regulates the inflammatory response in chronic rhinosinusitis with nasal polyps

Diagnostic Pathology ◽

10.1186/s13000-021-01089-0 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Jianwei Zhang ◽

Lei Han ◽

Feng Chen

Keyword(s):

Inflammatory Response ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Mapk Pathway ◽

Inhibitory Effect ◽

Luciferase Reporter ◽

Western Blot Assay ◽

Protein Levels ◽

Tnf Α

Abstract Background Let-7a-5p is demonstrated to be a tumor inhibitor in nasopharyngeal carcinoma. However, the role of let-7a-5p in chronic rhinosinusitis with nasal polyps (CRSwNP) has not been reported. This study is designed to determine the pattern of expression and role of let-7a-5p in CRSwNP. Methods The expression level of let-7a-5p, TNF-α, IL-1β, and IL-6 in CRSwNP tissues and cells were detected by RT-qPCR. Western blot assay was carried out to measure the protein expression of the Ras-MAPK pathway. Dual luciferase reporter assay and RNA pull-down assay were used to explore the relationship between let-7a-5p and IL-6. Results Let-7a-5p was significantly downregulated in CRSwNP tissues and cells. Moreover, the mRNA expression of TNF-α, IL-1β and IL-6 was increased in CRSwNP tissues, while let-7a-5p mimic inhibited the expression of TNF-α, IL-1β and IL-6. Besides that, let-7a-5p was negatively correlated with TNF-α, IL-1β and IL-6 in CRSwNP tissues. In our study, IL-6 was found to be a target gene of let-7a-5p. Additionally, let-7-5p mimic obviously reduced the protein levels of Ras, p-Raf1, p-MEK1 and p-ERK1/2, while IL-6 overexpression destroyed the inhibitory effect of let-7a-5p on the Ras-MAPK pathway in CRSwNP. Conclusion We demonstrated that let-7a-5p/IL-6 interaction regulated the inflammatory response through the Ras-MAPK pathway in CRSwNP.

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PBRM1 Cooperates with YTHDF2 to Control HIF-1α Protein Translation

Cells ◽

10.3390/cells10061425 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1425

Author(s):

Alena Shmakova ◽

Mark Frost ◽

Michael Batie ◽

Niall S. Kenneth ◽

Sonia Rocha

Keyword(s):

Protein Expression ◽

Cellular Response ◽

Transcriptional Activity ◽

Mrna Translation ◽

Protein Translation ◽

Signalling Pathways ◽

Independent Function ◽

Protein Levels ◽

Associated Proteins ◽

Core Components

PBRM1, a component of the chromatin remodeller SWI/SNF, is often deleted or mutated in human cancers, most prominently in renal cancers. Core components of the SWI/SNF complex have been shown to be important for the cellular response to hypoxia. Here, we investigated how PBRM1 controls HIF-1α activity. We found that PBRM1 is required for HIF-1α transcriptional activity and protein levels. Mechanistically, PBRM1 is important for HIF-1α mRNA translation, as absence of PBRM1 results in reduced actively translating HIF-1α mRNA. Interestingly, we found that PBRM1, but not BRG1, interacts with the m6A reader protein YTHDF2. HIF-1α mRNA is m6A-modified, bound by PBRM1 and YTHDF2. PBRM1 is necessary for YTHDF2 binding to HIF-1α mRNA and reduction of YTHDF2 results in reduced HIF-1α protein expression in cells. Our results identify a SWI/SNF-independent function for PBRM1, interacting with HIF-1α mRNA and the epitranscriptome machinery. Furthermore, our results suggest that the epitranscriptome-associated proteins play a role in the control of hypoxia signalling pathways.

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Application of Humanized Zebrafish Model in the Suppression of SARS-CoV-2 Spike Protein Induced Pathology by Tri-Herbal Medicine Coronil via Cytokine Modulation

Molecules ◽

10.3390/molecules25215091 ◽

2020 ◽

Vol 25 (21) ◽

pp. 5091

Author(s):

Acharya Balkrishna ◽

Siva Kumar Solleti ◽

Sudeep Verma ◽

Anurag Varshney

Keyword(s):

Herbal Medicine ◽

High Performance ◽

Normal Skin ◽

A549 Cells ◽

Lung Epithelial Cells ◽

Spike Protein ◽

Protective Effects ◽

Cell Degeneration ◽

Zebrafish Model ◽

Tnf Α

Zebrafish has been a reliable model system for studying human viral pathologies. SARS-CoV-2 viral infection has become a global chaos, affecting millions of people. There is an urgent need to contain the pandemic and develop reliable therapies. We report the use of a humanized zebrafish model, xeno-transplanted with human lung epithelial cells, A549, for studying the protective effects of a tri-herbal medicine Coronil. At human relevant doses of 12 and 58 µg/kg, Coronil inhibited SARS-CoV-2 spike protein, induced humanized zebrafish mortality, and rescued from behavioral fever. Morphological and cellular abnormalities along with granulocyte and macrophage accumulation in the swim bladder were restored to normal. Skin hemorrhage, renal cell degeneration, and necrosis were also significantly attenuated by Coronil treatment. Ultra-high-performance liquid chromatography (UHPLC) analysis identified ursolic acid, betulinic acid, withanone, withaferine A, withanoside IV–V, cordifolioside A, magnoflorine, rosmarinic acid, and palmatine as phyto-metabolites present in Coronil. In A549 cells, Coronil attenuated the IL-1β induced IL-6 and TNF-α cytokine secretions, and decreased TNF-α induced NF-κB/AP-1 transcriptional activity. Taken together, we show the disease modifying immunomodulatory properties of Coronil, at human equivalent doses, in rescuing the pathological features induced by the SARS-CoV-2 spike protein, suggesting its potential use in SARS-CoV-2 infectivity.

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