Sorting Nexin 10 Mediates Metabolic Reprogramming of Macrophages in Atherosclerosis Through the Lyn-Dependent TFEB Signaling Pathway

Rationale: SNX10 (sorting nexin 10) has been reported to play a critical role in regulating macrophage function and lipid metabolism. Objective: To investigate the precise role of SNX10 in atherosclerotic diseases and the underlying mechanisms. Methods and Results: SNX10 expression was compared between human healthy vessels and carotid atherosclerotic plaques. Myeloid cell-specific SNX10 knockdown mice were crossed onto the APOE −/− (apolipoprotein E) background and atherogenesis (high-cholesterol diet-induced) was monitored for 16 weeks. We found that SNX10 expression was increased in atherosclerotic lesions of aortic specimens from humans and APOE −/− mice. Myeloid cell-specific SNX10 deficiency (Δ knockout [KO]) attenuated atherosclerosis progression in APOE −/− mice. The population of anti-inflammatory monocytes/macrophages was increased in the peripheral blood and atherosclerotic lesions of ΔKO mice. In vitro experiments showed that SNX10 deficiency-inhibited foam cell formation through interrupting the internalization of CD36, which requires the interaction of SNX10 and Lyn-AKT (protein kinase B). The reduced Lyn-AKT activation by SNX10 deficiency promoted the nuclear translocation of TFEB (transcription factor EB), thereby enhanced lysosomal biogenesis and LAL (lysosomal acid lipase) activity, resulting in an increase of free fatty acids to fuel mitochondrial fatty acid oxidation. This further promoted the reprogramming of macrophages and shifted toward the anti-inflammatory phenotype. Conclusions: Our data demonstrate for the first time that SNX10 plays a crucial role in diet-induced atherogenesis via the previously unknown link between the Lyn-Akt-TFEB signaling pathway and macrophage reprogramming, suggest that SNX10 may be a potentially promising therapeutic target for atherosclerosis treatment.

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Ethanol Extract of Lilium Bulbs Plays an Anti-Inflammatory Role by Targeting the IKKα/β-Mediated NF-κB Pathway in Macrophages

The American Journal of Chinese Medicine ◽

10.1142/s0192415x18500672 ◽

2018 ◽

Vol 46 (06) ◽

pp. 1281-1296 ◽

Cited By ~ 9

Author(s):

Sang Yun Han ◽

Young-Su Yi ◽

Seong-Gu Jeong ◽

Yo Han Hong ◽

Kang Jun Choi ◽

...

Keyword(s):

Nitric Oxide ◽

Signaling Pathway ◽

Nuclear Translocation ◽

Inflammatory Responses ◽

Inflammatory Activity ◽

Raw264.7 Cells ◽

No Production ◽

Dependent Manner ◽

Bioactive Components ◽

Anti Inflammatory

Lilium bulbs have long been used as Chinese traditional medicines to alleviate the symptoms of various human inflammatory diseases. However, mechanisms of Lilium bulb-mediated anti-inflammatory activity and the bioactive components in Lilium bulbs remain unknown. In the present study, the anti-inflammatory activity of Lilium bulbs and the underlying mechanism of action were investigated in macrophages using Lilium bulb ethanol extracts (Lb-EE). In a dose-dependent manner, Lb-EE inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and bone marrow-derived macrophages (BMDMs) without causing significant cytotoxicity. Lb-EE also down-regulated mRNA expression of inflammatory genes in LPS-stimulated RAW264.7 cells, which included inducuble nitric oxide synthase (iNOS), cyclooxygenase-2 (COX2), and tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]). Furthermore, Lb-EE markedly restored LPS-induced morphological changes in RAW264.7 cells to a normal morphology. HPLC analysis identified quercetin, luteolin, and kaempferol as bioactive components contained in Lb-EE. Mechanistic studies in LPS-stimulated RAW264.7 cells revealed that Lb-EE suppressed MyD88- and TRIF-induced NF-[Formula: see text]B transcriptional activation and the nuclear translocation of NF-[Formula: see text]B transcription factors. Moreover, Lb-EE inhibited IKK[Formula: see text]/[Formula: see text]-induced activation of the NF-[Formula: see text]B signaling pathway and IKK inhibition significantly reduced NO production in LPS-stimulated RAW264.7 cells. Taken together, these results suggest that Lb-EE plays an anti-inflammatory role by targeting IKK[Formula: see text]/[Formula: see text]-mediated activation of the NF-[Formula: see text]B signaling pathway during macrophage-mediated inflammatory responses.

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Mettl14 Mediates the Inflammatory Response of Macrophages in Atherosclerosis Through the NF-κB/IL-6 Signaling Pathway

10.21203/rs.3.rs-1070642/v1 ◽

2021 ◽

Author(s):

Yang Zheng ◽

Yunqi Li ◽

Xianwen Ran ◽

Di Wang ◽

Xianghui Zheng ◽

...

Keyword(s):

Inflammatory Response ◽

Signaling Pathway ◽

Foam Cell ◽

Epigenetic Modification ◽

Gene Knockout ◽

Critical Role ◽

Cell Formation ◽

Vital Role ◽

Foam Cell Formation

Abstract The inflammatory response of macrophages has been reported to play a critical role in atherosclerosis. The inflammatory state of macrophages is modified by epigenetic reprogramming. m6A RNA methylation is an epigenetic modification of RNAs. However, little is known about the potential roles and underlying mechanisms of m6A modification in macrophage inflammation. Herein, we showed that the expression of the m6A modification “writer” Mettl14 was increased in coronary heart disease and LPS-stimulated THP-1 cells. Knockdown of Mettl14 promoted M2 polarization of macrophages, inhibited foam cell formation and decreased migration. Mechanistically, the expression of Myd88 and IL-6 was decreased in Mettl14 knockdown cells. Through m6A modification, Mettl14 regulated the stability of Myd88 mRNA. Furthermore, Myd88 affected the transcription of IL-6 via the distribution of p65 in nuclei rather than directly regulating the expression of IL-6 through m6A modification. In vivo, Mettl14 gene knockout significantly reduced the inflammatory response of macrophages and the development of atherosclerotic plaques. Taken together, our data demonstrate that Mettl14 plays a vital role in macrophage inflammation in atherosclerosis via the NF-κB/IL-6 signaling pathway, suggesting that Mettl14 may be a promising therapeutic target for the clinical treatment of atherosclerosis.

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Berry Phenolic and Volatile Extracts Inhibit Pro-Inflammatory Cytokine Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-κB Signaling Pathway

Antioxidants ◽

10.3390/antiox9090871 ◽

2020 ◽

Vol 9 (9) ◽

pp. 871

Author(s):

Inah Gu ◽

Cindi Brownmiller ◽

Nathan B. Stebbins ◽

Andy Mauromoustakos ◽

Luke Howard ◽

...

Keyword(s):

Signaling Pathway ◽

Nuclear Translocation ◽

Complex Mixture ◽

Limited Information ◽

Potential Health ◽

Tnf Α ◽

Anti Inflammatory ◽

Factor Α ◽

Raw264.7 Macrophage ◽

Necrosis Factor

Berries are a rich source of phytochemicals, especially phenolics well known for protective activity against many chronic diseases. Berries also contain a complex mixture of volatile compounds that are responsible for the unique aromas of berries. However, there is very limited information on the composition and potential health benefits of berry volatiles. In this study, we isolated phenolic and volatile fractions from six common berries and characterized them by HPLC/HPLC-MS and GC/GC-MS, respectively. Berry phenolic and volatile fractions were evaluated for an anti-inflammatory effect using lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells by measuring levels of pro-inflammatory cytokines and the nuclear factor-kappa B (NF-κB) signaling pathway. Results showed that LPS-induced excessive production of nitric oxide (NO), prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), which were inhibited by berry phenolic and volatile extracts. Moreover, berry phenolic and volatile extracts reduced the nuclear translocation of NF-κB by blocking the phosphorylation of p65 and degradation of IκBα. These findings showed that berry volatiles from six berries had comparable anti-inflammatory effects to berry phenolics through the suppression of pro-inflammatory mediators and cytokines expression via NF-κB down-regulation, despite being present in the fruit at a lower concentration.

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Arsenic trioxide induces macrophage autophagy and atheroprotection by regulating ROS-dependent TFEB nuclear translocation and AKT/mTOR pathway

Cell Death and Disease ◽

10.1038/s41419-020-03357-1 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Shaohong Fang ◽

Xin Wan ◽

Xiaoyi Zou ◽

Song Sun ◽

Xinran Hao ◽

...

Keyword(s):

Arsenic Trioxide ◽

Signaling Pathway ◽

Nuclear Translocation ◽

Early Stage ◽

Mtor Pathway ◽

Mtor Signaling ◽

Mtor Signaling Pathway ◽

Pi3k Inhibitor Ly294002 ◽

Vascular Stents ◽

Atherosclerotic Lesions

AbstractInducing autophagy and inhibiting apoptosis may provide a therapeutic treatment for atherosclerosis (AS). For the treatment of progressive AS, arsenic trioxide (ATO) has been used to coat vascular stents. However, the effect of ATO on autophagy of macrophages is still unknown. Therefore, the aims of this study were to characterize the effects and the mechanism of actions of ATO on autophagy in macrophages. Our results showed that ATO-induced activation of autophagy was an earlier event than ATO-induced inhibition of the expression of apoptosis markers in macrophages and foam cells. Nuclear transcription factor EB (TFEB) prevents atherosclerosis by activating macrophage autophagy and promoting lysosomal biogenesis. Here, we report that ATO triggered the nuclear translocation of TFEB, which in turn promoted autophagy and autophagosome-lysosome fusion. Both the latter events were prevented by TFEB knockdown. Moreover, ATO decreased the p-AKT and p-mTOR in the PI3K/AKT/mTOR signaling pathway, thus inducing autophagy. Correspondingly, treatment with the autophagy inhibitor 3-methyladenine (3-MA) abolished the autophagy-inducing effects of ATO. Meanwhile, PI3K inhibitor (LY294002) and mTOR inhibitor (rapamycin) cooperated with ATO to induce autophagy. Furthermore, reactive oxygen species (ROS) were generated in macrophages after treatment with ATO. The ROS scavenger N-acetyl-1-cysteine (NAC) abolished ATO-induced nuclear translocation of TFEB, as well as changes in key molecules of the AKT/mTOR signaling pathway and downstream autophagy. More importantly, ATO promoted autophagy in the aorta of ApoE−/− mice and reduced atherosclerotic lesions in early AS, which were reversed by 3-MA treatment. In summary, our data indicated that ATO promoted ROS induction, which resulted in nuclear translocation of TFEB and inhibition of the PI3K/AKT/mTOR pathway. These actions ultimately promoted macrophage autophagy and reduced atherosclerotic lesions at early stages. These findings may provide a new perspective for the clinical treatment of early-stage atherosclerosis and should be further studied.

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Macrophage-derived miR-21 drives overwhelming glycolytic and inflammatory response during sepsis via repression of the PGE2/IL-10 axis

10.1101/2020.05.09.086421 ◽

2020 ◽

Author(s):

Paulo Melo ◽

Annie Rocio Pineros Alvarez ◽

C. Henrique Serezani

Keyword(s):

Inflammatory Response ◽

Systemic Inflammation ◽

Inflammatory Mediators ◽

Aerobic Glycolysis ◽

Critical Role ◽

Organ Damage ◽

Metabolic Reprogramming ◽

Prostaglandin E ◽

Anti Inflammatory

AbstractMyeloid cells play a critical role in the development of systemic inflammation and organ damage during sepsis. The mechanisms the development of aberrant inflammatory response remains to be elucidated. MicroRNAs are small non-coding RNAs that could prevent the expression of inflammatory molecules. Although the microRNA-21 (miR-21) is abundantly expressed in macrophages, the role of miR-21 in sepsis is controversial. Here we showed that miR-21 is upregulated in neutrophils and macrophages from septic mice. We found that myeloid-specific miR-21 deletion enhances animal survival, followed by decreased bacterial growth and organ damage during sepsis. Increased resistance against sepsis was associated with a reduction of aerobic glycolysis (as determined by reduced extracellular acidification rate (ECAR) and expression of glycolytic enzymes) and systemic inflammatory response (IL-1βTNFα and IL-6). While miR-21-/- macrophages failed to induce aerobic glycolysis and production of pro-inflammatory cytokines, we observed increased levels of the anti-inflammatory mediators’ prostaglandin E2 (PGE2) and IL10. Using blocking antibodies and pharmacological tools, we further discovered that increased survival and decreased systemic inflammation in miR21Δmyel during sepsis is dependent on the PGE2/IL10-mediated glycolysis inhibition. Together, we are showing a heretofore unknown role of macrophage miR21 in the orchestrating the balance between anti-inflammatory mediators and metabolic reprogramming that drives cytokine storm and tissue damage during sepsis.

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TGF-βRII regulates glucose metabolism in oral cancer-associated fibroblasts via promoting PKM2 nuclear translocation

Cell Death Discovery ◽

10.1038/s41420-021-00804-6 ◽

2022 ◽

Vol 8 (1) ◽

Author(s):

Fanglong Wu ◽

Shimeng Wang ◽

Qingxiang Zeng ◽

Junjiang Liu ◽

Jin Yang ◽

...

Keyword(s):

Glucose Metabolism ◽

Oral Cancer ◽

Nuclear Translocation ◽

Aerobic Glycolysis ◽

Critical Role ◽

Metabolic Reprogramming ◽

Cancer Associated Fibroblasts

AbstractCancer-associated fibroblasts (CAFs) are highly heterogeneous and differentiated stromal cells that promote tumor progression via remodeling of extracellular matrix, maintenance of stemness, angiogenesis, and modulation of tumor metabolism. Aerobic glycolysis is characterized by an increased uptake of glucose for conversion into lactate under sufficient oxygen conditions, and this metabolic process occurs at the site of energy exchange between CAFs and cancer cells. As a hallmark of cancer, metabolic reprogramming of CAFs is defined as reverse Warburg effect (RWE), characterized by increased lactate, glutamine, and pyruvate, etc. derived from aerobic glycolysis. Given that the TGF-β signal cascade plays a critical role in RWE mainly through metabolic reprogramming related proteins including pyruvate kinase muscle isozyme 2 (PKM2), however, the role of nuclear PKM2 in modifying glycolysis remains largely unknown. In this study, using a series of in vitro and in vivo experiments, we provide evidence that TGF-βRII overexpression suppresses glucose metabolism in CAFs by attenuating PKM2 nuclear translocation, thereby inhibiting oral cancer tumor growth. This study highlights a novel pathway that explains the role of TGF-βRII in CAFs glucose metabolism and suggests that targeting TGF-βRII in CAFs might represent a therapeutic approach for oral cancer.

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PCSK9 and LRP5 in macrophage lipid internalization and inflammation

Cardiovascular Research ◽

10.1093/cvr/cvaa254 ◽

2020 ◽

Cited By ~ 1

Author(s):

Lina Badimon ◽

Aureli Luquero ◽

Javier Crespo ◽

Esther Peña ◽

Maria Borrell-Pages

Keyword(s):

Nuclear Translocation ◽

Foam Cell ◽

Critical Role ◽

Primary Cultures ◽

Cell Formation ◽

Density Lipoprotein ◽

Blood Cholesterol ◽

Foam Cell Formation ◽

Lipid Uptake ◽

Cholesterol Levels

Abstract Aims Atherosclerosis, the leading cause of cardiovascular diseases, is driven by high blood cholesterol levels and chronic inflammation. Low-density lipoprotein receptors (LDLR) play a critical role in regulating blood cholesterol levels by binding to and clearing LDLs from the circulation. The disruption of the interaction between proprotein convertase subtilisin/kexin 9 (PCSK9) and LDLR reduces blood cholesterol levels. It is not well known whether other members of the LDLR superfamily may be targets of PCSK9. The aim of this work was to determine if LDLR-related protein 5 (LRP5) is a PCSK9 target and to study the role of PCSK9 and LRP5 in foam cell formation and lipid accumulation. Methods and results Primary cultures of human inflammatory cells (monocytes and macrophages) were silenced for LRP5 or PCSK9 and challenged with LDLs. We first show that LRP5 is needed for macrophage lipid uptake since LRP5-silenced macrophages show less intracellular CE accumulation. In macrophages, internalization of LRP5-bound LDL is already highly evident after 5 h of LDL incubation and lasts up to 24 h; however, in the absence of both LRP5 and PCSK9, there is a strong reduction of CE accumulation indicating a role for both proteins in lipid uptake. Immunoprecipitation experiments show that LRP5 forms a complex with PCSK9 in lipid-loaded macrophages. Finally, PCSK9 participates in TLR4/NFkB signalling; a decreased TLR4 protein expression levels and a decreased nuclear translocation of NFκB were detected in PCSK9 silenced cells after lipid loading, indicating a downregulation of the TLR4/NFκB pathway. Conclusion Our results show that both LRP5 and PCSK9 participate in lipid uptake in macrophages. In the absence of LRP5, there is a reduced release of PCSK9 indicating that LRP5 also participates in the mechanism of release of soluble PCSK9. Furthermore, PCSK9 up-regulates TLR4/NFκB favouring inflammation.

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Gambogic acid, a novel ligand for transferrin receptor, potentiates TNF-induced apoptosis through modulation of the nuclear factor-κB signaling pathway

Blood ◽

10.1182/blood-2007-03-079616 ◽

2007 ◽

Vol 110 (10) ◽

pp. 3517-3525 ◽

Cited By ~ 162

Author(s):

Manoj K. Pandey ◽

Bokyung Sung ◽

Kwang Seok Ahn ◽

Ajaikumar B. Kunnumakkara ◽

Madan M. Chaturvedi ◽

...

Keyword(s):

Signaling Pathway ◽

Transferrin Receptor ◽

Nuclear Translocation ◽

Critical Role ◽

Nuclear Factor Κb ◽

Chemotherapeutic Agents ◽

Gambogic Acid ◽

Human Leukemia ◽

Gene Products ◽

Signaling Mechanism

Abstract Gambogic acid (GA), a xanthone derived from the resin of the Garcinia hanburyi, has been recently demonstrated to bind transferrin receptor and exhibit potential anticancer effects through a signaling mechanism that is not fully understood. Because of the critical role of NF-κB signaling pathway, we investigated the effects of GA on NF-κB–mediated cellular responses and NF-κB–regulated gene products in human leukemia cancer cells. Treatment of cells with GA enhanced apoptosis induced by tumor necrosis factor (TNF) and chemotherapeutic agents, inhibited the expression of gene products involved in antiapoptosis (IAP1 and IAP2, Bcl-2, Bcl-xL, and TRAF1), proliferation (cyclin D1 and c-Myc), invasion (COX-2 and MMP-9), and angiogenesis (VEGF), all of which are known to be regulated by NF-κB. GA suppressed NF-κB activation induced by various inflammatory agents and carcinogens and this, accompanied by the inhibition of TAK1/TAB1-mediated IKK activation, inhibited IκBα phosphorylation and degradation, suppressed p65 phosphorylation and nuclear translocation, and finally abrogated NF-κB–dependent reporter gene expression. The NF-κB activation induced by TNFR1, TRADD, TRAF2, NIK, TAK1/TAB1, and IKKβ was also inhibited. The effect of GA mediated through transferrin receptor as down-regulation of the receptor by RNA interference reversed its effects on NF-κB and apoptosis. Overall our results demonstrate that GA inhibits NF-κB signaling pathway and potentiates apoptosis through its interaction with the transferrin receptor.

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Overexpressed WDR3 induces the activation of Hippo pathway by interacting with GATA4 in pancreatic cancer

10.21203/rs.3.rs-104564/v2 ◽

2021 ◽

Author(s):

Wenjie Su ◽

Shikai Zhu ◽

Kai Chen ◽

Hongji Yang ◽

Mingwu Tian ◽

...

Keyword(s):

Pancreatic Cancer ◽

Signaling Pathway ◽

Nuclear Translocation ◽

Critical Role ◽

Biological Role ◽

Hippo Signaling ◽

Hippo Signaling Pathway ◽

Pancreatic Cancer Cells ◽

Independent Functions

Abstract Background: WD repeat domain 3 (WDR3) is involved in a variety of cellular processes including gene regulation, cell cycle progression, signal transduction and apoptosis. However, the biological role of WDR3 in pancreatic cancer and the associated mechanism remains unclear. We seek to explore the immune-independent functions and relevant mechanism for WDR3 in pancreatic cancer.Methods: The GEPIA web tool was searched, and IHC assays were conducted to determine the mRNA and protein expression levels of WDR3 in pancreatic cancer patients. MTS, colony formation, and transwell assays were conducted to determine the biological role of WDR3 in human cancer. Western blot analysis, RT-qPCR, and immunohistochemistry were used to detect the expression of specific genes. An immunoprecipitation assay was used to explore protein-protein interactions.Results: Our study proved that overexpressed WDR3 was correlated with poor survival in pancreatic cancer and that WDR3 silencing significantly inhibited the proliferation, invasion, and tumor growth of pancreatic cancer. Furthermore, WDR3 activated the Hippo signaling pathway by inducing yes association protein 1 (YAP1) expression, and the combination of WDR3 silencing and administration of the YAP1 inhibitor TED-347 had a synergistic inhibitory effect on the progression of pancreatic cancer. Finally, the upregulation of YAP1 expression induced by WDR3 was dependent on an interaction with GATA binding protein 4 (GATA4), the transcription factor of YAP1, which interaction induced the nuclear translocation of GATA4 in pancreatic cancer cells.Conclusions: We identified a novel mechanism by which WDR3 plays a critical role in promoting pancreatic cancer progression by activating the Hippo signaling pathway through the interaction with GATA4. Therefore, WDR3 is potentially a therapeutic target for pancreatic cancer treatment.

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Bergenin-activated SIRT1 inhibits TNF-a-induced pro-inflammatory response via blocking NF-kB signaling pathway

10.21203/rs.2.20671/v1 ◽

2020 ◽

Author(s):

Bei xian Zhou ◽

Bin Wu ◽

Min Chen ◽

Cuifen Chen ◽

Yun Gao ◽

...

Keyword(s):

Inflammatory Response ◽

Signaling Pathway ◽

Western Blotting ◽

Nuclear Translocation ◽

Sirtuin 1 ◽

Burn Wound ◽

Burn Wound Healing ◽

Mrna And Protein Expression ◽

Sirt1 Inhibitor ◽

Anti Inflammatory

Abstract BackgroundBergenin, a kind of polyphenol compound, has been showed to exhibit antiulcerogenic, anti-inflammatory, antitussive and burn wound healing properties, However, its therapeutic effect on TNF-a induced pro-inflammatory response in airway and its potential mechanisms of actions were still unclearAim of this studyThis study aimed to investigate the anti-inflammatory effects and possible mechanism of bergenin in TNF-a-stimulated 16-HBE cells.Materials and methodsCCK8 was used to determine the cytotoxicity. Cytokines expression were analyzed by qRT-qPCR and ELISA. Immunofluorescence, western blotting and SIRT1 activity assay were employed to investigate the possible molecular mechanism. ResultsOur results showed that bergenin treatment obviously decreased the mRNA and protein expressions of IL-6 and IL-8 in TNF-a-stimulated 16-HBE cells. Bergenin blocked TNF-a-mediated the activation of NF-kB signaling and NF-kB nuclear translocation. Interestingly, RT‑qPCR and western blotting revealed that bergenin did not affect the expression of sirtuin-1 (SIRT1), but significantly increased the activity of SIRT1. Furthermore, bergenin-mediated SIRT1 activation was further confirmed by the resulted that bergenin decreased acetylation levels of NF-kB-p65 and p53. The inhibitory effects of bergenin on the mRNA and protein expression levels of IL-6 and IL-8 were reversed by addition of SIRT1 inhibitor. In addition, combination of bergenin and dexamethasone showed additive effects on the reduction of IL-6 and IL-8.ConclusionsThese findings demonstrated that bergenin could suppress TNF-α-induced pro-inflammatory response via augmenting SIRT1 activity to block NF-kB signaling pathway, which may contribute to provide beneficial effect for the treatment of airway inflammation in asthma.

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