bims-noxint 2020-03-15 papers

bims-noxint

Biomed News

on NADPH oxidases in tumorigenesis

Issue of 2020–03–15
nine papers selected by
Laia Caja Puigsubira, Uppsala University

High-fat diet increased NADPH-oxidase-related oxidative stress and aggravated LPS-induced intestine injury.
Role of oxidative stress in the pathogenesis of nonalcoholic fatty liver disease.
Induction of Cyclooxygenase-2 by Overexpression of the Human NADPH Oxidase 5 (NOX5) Gene in Aortic Endothelial Cells.
Effect of beta- and alpha-glucans on immune modulating factors expression in enterocyte-like Caco-2 and goblet-like LS 174T cells.
NOX-dependent reactive oxygen species production underlies arrhythmias susceptibility in dexamethasone-treated rats.
Reduced EGFR Level in eIF2α Phosphorylation-Deficient Hepatocytes Is Responsible for Susceptibility to Oxidative Stress.
NoxO1 Knockout Promotes Longevity in Mice.
Muscone relieves inflammatory pain by inhibiting microglial activation-mediated inflammatory response via abrogation of the NOX4/JAK2-STAT3 pathway and NLRP3 inflammasome.
Identification of highly potent and selective inhibitor, TIPTP, of the p22phox-Rubicon axis as a therapeutic agent for rheumatoid arthritis.

Life Sci. 2020 Mar 09. pii: S0024-3205(20)30287-3. [Epub ahead of print] 117539

High-fat diet increased NADPH-oxidase-related oxidative stress and aggravated LPS-induced intestine injury.

Shan Wu, Lijie Pan, Haofeng Liao, Weifeng Yao, Ning Shen, Chaojin Chen, Dezhao Liu, Mian Ge.

   AIMS: Lipopolysaccharide (LPS)-induced intestinal injury is a common clinical feature of sepsis. Aggravated inflammation and higher sensitivity to infection are associated with high-fat diet (HFD) in patients with type 2 diabetes and/or obesity. However, the mechanism by which HFD exacerbates LPS-induced intestinal injury has not been elucidated. This study aims to examine the effects of HFD on intestinal injury induced by LPS and the underlying mechanism.
MAIN METHODS: Mice were fed with HFD or regular chow for 12 weeks and were then challenged with LPS. Vas2870 was administered to mice that received HFD before the initiation of the diet. The levels of tight junction protein expression, oxidative stress, organ injury, and nicotinamide adenine dinucleotide phosphate (NADPH)-associated proteins were assessed periodically.
KEY FINDINGS: LPS treatment resulted in severe intestinal pathological injury and increased oxidative stress, evidenced by significantly increased serum diamine oxidase, reactive oxygen species, malondialdehyde, and intestinal fatty acid binding protein contents. Additionally, a decrease in tight junction protein expression was observed, indicating a loss of tight junction integrity. LPS treatment induced the expression of Nox2 and Nox4. All the effects were more severe in HFD mice. Treatment with vas2870 conferred protection against LPS-induced intestinal injury in HFD-fed mice, partially reduced oxidative stress, and rescued the expression of tight junction proteins.
CONCLUSION: HFD aggravated LPS-induced intestine injury through exacerbating intestinal Nox-related oxidative stress, which led to a loss of the integrity of tight junctions and consequently increased intestinal permeability.

Keywords:  High fat diet; LPS-induced intestinal injury; NADPH oxidases

DOI:  https://doi.org/10.1016/j.lfs.2020.117539
Free Radic Biol Med. 2020 Mar 07. pii: S0891-5849(19)31515-1. [Epub ahead of print]

Role of oxidative stress in the pathogenesis of nonalcoholic fatty liver disease.

Ze Chen, Ruifeng Tian, Zhigang She, Jingjing Cai, Hongliang Li.

  Nonalcoholic fatty liver disease (NAFLD) has emerged as the most common chronic liver disease worldwide and is strongly associated with the presence of oxidative stress. Disturbances in lipid metabolism lead to hepatic lipid accumulation, which affects different reactive oxygen species (ROS) generators, including mitochondria, endoplasmic reticulum, and NADPH oxidase. Mitochondrial function adapts to NAFLD mainly through the downregulation of the electron transport chain (ETC) and the preserved or enhanced capacity of mitochondrial fatty acid oxidation, which stimulates ROS overproduction within different ETC components upstream of cytochrome c oxidase. However, non-ETC sources of ROS, in particular, fatty acid β-oxidation, appear to produce more ROS in hepatic metabolic diseases. Endoplasmic reticulum stress and NADPH oxidase alterations are also associated with NAFLD, but the degree of their contribution to oxidative stress in NAFLD remains unclear. Increased ROS generation induces changes in insulin sensitivity and in the expression and activity of key enzymes involved in lipid metabolism. Moreover, the interaction between redox signaling and innate immune signaling forms a complex network that regulates inflammatory responses. Based on the mechanistic view described above, this review summarizes the mechanisms that may account for the excessive production of ROS, the potential mechanistic roles of ROS that drive NAFLD progression, and therapeutic interventions that are related to oxidative stress.

Keywords:  Endoplasmic reticulum stress; Innate immunity; Insulin resistance; Lipid metabolism; Mitochondrial dysfunction; Nonalcoholic fatty liver disease; Oxidative stress

DOI:  https://doi.org/10.1016/j.freeradbiomed.2020.02.025
Cells. 2020 Mar 06. pii: E637. [Epub ahead of print]9(3):

Induction of Cyclooxygenase-2 by Overexpression of the Human NADPH Oxidase 5 (NOX5) Gene in Aortic Endothelial Cells.

Javier Marqués, Adriana Cortés, Álvaro Pejenaute, Eduardo Ansorena, Gloria Abizanda, Felipe Prósper, Juan José Martínez-Irujo, Carlos de Miguel, Guillermo Zalba.

  Oxidative stress is a main molecular mechanism that underlies cardiovascular diseases. A close relationship between reactive oxygen species (ROS) derived from NADPH oxidase (NOX) activity and the prostaglandin (PG) biosynthesis pathway has been described. However, little information is available about the interaction between NOX5 homolog-derived ROS and the PG pathway in the cardiovascular context. Our main goal was to characterize NOX5-derived ROS effects in PG homeostasis and their potential relevance in cardiovascular pathologies. For that purpose, two experimental systems were employed: an adenoviral NOX5-β overexpression model in immortalized human aortic endothelial cells (TeloHAEC) and a chronic infarction in vivo model developed from a conditional endothelial NOX5 knock-in mouse. NOX5 increased cyclooxygenase-2 isoform (COX-2) expression and prostaglandin E2 (PGE2) production through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in TeloHAEC. Protein kinase C (PKC) activation and intracellular calcium level (Ca++) mobilization increased ROS production and NOX5 overexpression, which promoted a COX-2/PGE2 response in vitro. In the chronic infarction model, mice encoding endothelial NOX5 enhanced the cardiac mRNA expression of COX-2 and PGES, suggesting a COX-2/PGE2 response to NOX5 presence in an ischemic situation. Our data support that NOX5-derived ROS may modulate the COX-2/PGE2 axis in endothelial cells, which might play a relevant role in the pathophysiology of heart infarction.

Keywords:  COX-2; NADPH oxidase 5; NF-κB; PGE2; PKC; chronic infarction; oxidative stress

DOI:  https://doi.org/10.3390/cells9030637
Int J Biol Macromol. 2020 Mar 09. pii: S0141-8130(19)39856-3. [Epub ahead of print]

Effect of beta- and alpha-glucans on immune modulating factors expression in enterocyte-like Caco-2 and goblet-like LS 174T cells.

Bruna De Felice, Simona Damiano, Concetta Montanino, Andrea Del Buono, Giuliana La Rosa, Bruna Guida, Mariarosaria Santillo.

  Glucans are complex polysaccharides consisting of repeated units of d-glucose linked by glycosidic bonds. The nutritional contribution in α-glucans is mainly given by starch and glycogen while in β-glucans by mushrooms, yeasts and whole grains, such as barley and spelt well represented in the Mediterranean Diet. Numerous and extensive studies performed on glucans highlighted their marked anti-tumor, antioxidant and immunomodulatory activity. It has recently been shown that rather than merely being a passive barrier, the intestinal epithelium is an essential modulator of immunity. Indeed, epithelial absorptive enterocytes and mucin secreting goblet cells can produce specific immune modulating factors, driving innate immunity to pathogens as well as preventing autoimmunity. Despite the clear evidence of the effects of glucans on immune system cells, there are only limited data about their effects on immune activity of mucosal intestinal cells strictly related to intestinal barrier integrity. The aim of the study was to evaluate the effects of α and β glucans, alone or in combination with other substances with antioxidant properties, on reactive oxygen species (ROS) levels, on the expression of ROS-generating enzyme DUOX-2 and of the immune modulating factors Tumor Necrosis Factor (TNF-α), Interleukin 1 β (IL-1β) and cyclooxygenase-2 (COX-2) in two intestinal epithelial cells, the enterocyte-like Caco-2 cells and goblet cell-like LS174T. In our research, the experiments were carried out incubating the cells with glucans for 18 h in culture medium containing 0.2% FBS and measuring ROS levels fluorimetrically as dihydrodichlorofluoresce diacetate (DCF-DA) fluorescence, protein levels of DUOX-2 by Western blotting and mRNA levels of, TNF-α, IL-1β and COX-2 by qRT-PCR. α and β glucans decreased ROS levels in Caco-2 and LS 174T cells. The expression levels of COX-2, TNF-α, and IL-1β were also reduced by α- and β-glucans. Additive effects on the expression of these immune modulating factors were exerted by vitamin C. In Caco-2 cells, the dual oxidase DUOX-2 expression is positively modulated by ROS. Accordingly, in Caco-2 or LS174T cells treated with α and β-glucans alone or in combination with Vitamin C, the decrease of ROS levels was associated with a reduced expression of DUOX-2. The treatment of cells with the NADPH oxidase (NOX) inhibitor apocynin decrease ROS, DUOX-2, COX-2, TNF-α and IL-1β levels indicating that NOX dependent ROS regulate the expression of immune modulating factors of intestinal cells. However, the combination of vitamin C, α and β-glucans with apocynin did not exert an additive effect on COX-2, TNF-α and IL-1β levels when compared with α-, β-glucans and Vitamin C alone. The present study showing a modulatory effect of α and β-glucans on ROS and on the expression of immune modulating factors in intestinal epithelial cells suggests that the assumption of food containing high levels of these substances or dietary supplementation can contribute to normal immunomodulatory function of intestinal barrier.

Keywords:  Cyclooxygenase-2 (COX-2); Interleukin 1 β (IL-1β); Oxidative stress; ROS; Tumor necrosis factor (TNF-α); α and β-glucans

DOI:  https://doi.org/10.1016/j.ijbiomac.2020.03.046
Free Radic Biol Med. 2020 Mar 05. pii: S0891-5849(20)30407-X. [Epub ahead of print]152 1-7

NOX-dependent reactive oxygen species production underlies arrhythmias susceptibility in dexamethasone-treated rats.

Fabricio Nunes Macedo, Diego Santos de Souza, João Eliakim Dos Santos Araújo, Cácia Oliveira Dantas, Rodrigo Miguel-Dos-Santos, Elizabete Silva-Filha, Thallita Kelly Rabelo, Robervan Vidal Dos Santos, Rui Zhang, André Sales Barreto, Carla Maria Lins de Vasconcelos, Sandra Lauton-Santos, Márcio Roberto Viana Dos Santos, Lucindo José Quintans-Júnior, Valter Joviniano Santana-Filho, Thássio Ricardo Ribeiro Mesquita.

  Dexamethasone is the most clinically used glucocorticoid with an established role in the treatment of a wide spectrum of inflammatory-related diseases. While the therapeutic actions are well known, dexamethasone treatment causes a number of cardiovascular side effects, which are complex, frequent and, in some cases, clinically unnoticeable. Here, we investigated whether a therapeutic regimen of dexamethasone affects cardiac arrhythmogenesis, focusing on the contribution of Nox-derived reactive oxygen species (ROS). Male Wistar rats were treated with dexamethasone (2 mg/kg, i.p.) for 7 days. Afterward, hemodynamic measurements, autonomic modulation, left ventricular function, cardiac fibrosis, reactive oxygen species (ROS) generation, Nox protein expression, superoxide dismutase (SOD) and catalase activities, and arrhythmias incidence were evaluated. Here, we show that dexamethasone increases blood pressure, associated with enhanced cardiac and vascular sympathetic modulation. Moreover, a marked increase in the cardiac ROS generation was observed, whereas the enhanced SOD activity did not prevent the higher levels of lipid peroxidation in the dexamethasone group. On the other hand, increased cardiac Nox 4 expression and hydrogen peroxide decomposition rate was observed in dexamethasone-treated rats, while Nox 2 remained unchanged. Interestingly, although preserved ventricular contractility and β-adrenergic responsiveness, we found that dexamethasone-treated rats displayed greater interstitial and perivascular fibrosis than control. Surprisingly, despite the absence of arrhythmias at basal condition, we demonstrated, by in vivo and ex vivo approaches, that dexamethasone-treated rats are more susceptible to develop harmful forms of ventricular arrhythmias when challenged with pharmacological drugs or burst pacing-induced arrhythmias. Notably, concomitant treatment with apocynin, an inhibitor of NADPH oxidase, prevented these ectopic ventricular events. Together, our results reveal that hearts become arrhythmogenic during dexamethasone treatment, uncovering the pivotal role of ROS-generating NADPH oxidases for arrhythmias vulnerability.

Keywords:  Arrhythmia; Autonomic nervous system; Dexamethasone; Hypertension; NADPH oxidase

DOI:  https://doi.org/10.1016/j.freeradbiomed.2020.03.005
Mol Cells. 2020 Mar 09.

Reduced EGFR Level in eIF2α Phosphorylation-Deficient Hepatocytes Is Responsible for Susceptibility to Oxidative Stress.

Mi-Jeong Kim, Woo-Gyun Choi, Kyung-Ju Ahn, In Gyeong Chae, Rina Yu, Sung Hoon Back.

  Reactive oxygen species (ROS) play a significant role in intracellular signaling and regulation, particularly when they are maintained at physiologic levels. However, excess of ROS can cause cell damage and induce cell death. We recently reported that eIF2α phosphorylation protects hepatocytes from oxidative stress and liver fibrosis induced by fructose metabolism. Here, we found that hepatocyte-specific eIF2α phosphorylation-deficient mice have significantly reduced expression of the epidermal growth factor receptor (EGFR) and altered EGFR-mediated signaling pathways. EGFR-mediated signaling pathways are important for cell proliferation, differentiation, and survival in many tissues and cell types. Therefore, we studied whether the reduced amount of EGFR is responsible for the eIF2α phosphorylationdeficient hepatocytes' vulnerability to oxidative stress. ROS such as hydrogen peroxide and superoxides induce both EGFR tyrosine phosphorylation and eIF2α phosphorylation. eIF2α phosphorylation-deficient primary hepatocytes, or EGFR knockdown cells, have decreased ROS scavenging ability compared to normal cells. Therefore, these cells are particularly susceptible to oxidative stress. However, overexpression of EGFR in these eIF2α phosphorylationdeficient primary hepatocytes increased ROS scavenging ability and alleviated ROS-mediated cell death. Therefore, we hypothesize that the reduced EGFR level in eIF2α phosphorylation-deficient hepatocytes is one of critical factors responsible for their susceptibility to oxidative stress.

Keywords:  eIF2α phos phorylation; epidermal growth factor receptor; hydrogen peroxide; menadione; reactive oxygen species

DOI:  https://doi.org/10.14348/molcells.2020.2197
Antioxidants (Basel). 2020 Mar 10. pii: E226. [Epub ahead of print]9(3):

NoxO1 Knockout Promotes Longevity in Mice.

Tim Schader, Christina Reschke, Manuela Spaeth, Susanne Wienstroer, Szeka Wong, Katrin Schröder.

  According to the free radical theory of aging, reactive oxygen species (ROS) have been proposed to be a major cause of aging for a long time. Meanwhile, it became clear that ROS have diverse functions in a healthy organism. They act as second messengers, and as transient inhibitors of phosphatases and others. In fact, their detrimental role is highly dependent on the context of their production. NADPH oxidases (Nox) have been discovered as a controllable source of ROS. NoxO1 enables constitutive ROS formation by Nox1 by acting as a constitutively active cytosolic subunit of the complex. We previously found that both Nox1 and NoxO1 were highly expressed in the colon, and that NoxO1-/- deficiency reduces colon health. We hypothesized that a healthy colon potentially contributes to longevity and NoxO1 deficiency would reduce lifetime, at least in mouse. In contrast, here we provide evidence that the knockout of NoxO1 results in an elongated life expectancy of mice. No better endothelial function, nor an improved expression of genes related to longevity, such as Sirt1, were found, and therefore may not serve as an explanation for a longer life in NoxO1 deficiency. Rather minor systemic differences, such as lower body weight occur. As a potential reason for longer life, we suggest better DNA repair capacity in NoxO1 deficient mice. Although final fatal DNA damage appears similar between wildtype and NoxO1 knockout animals, we identified less intermediate DNA damage in colon cells of NoxO1-/- mice, while the number of cells with intact DNA is elevated in NoxO1-/- colons. We conclude that NoxO1 deficiency prolongs lifetime of mice, which correlates with less intermediate and potentially fixable DNA damage at least in colon cells.

Keywords:  NADPH oxidase; NoxO1; longevity; mice

DOI:  https://doi.org/10.3390/antiox9030226
Int Immunopharmacol. 2020 Mar 04. pii: S1567-5769(19)32684-0. [Epub ahead of print]82 106355

Muscone relieves inflammatory pain by inhibiting microglial activation-mediated inflammatory response via abrogation of the NOX4/JAK2-STAT3 pathway and NLRP3 inflammasome.

Shanshan Yu, Guoqing Zhao, Fanglei Han, Wenzhao Liang, Yuan Jiao, Zinan Li, Longyun Li.

  Previous studies have shown that muscone, a pharmacologically active ingredient isolated from musk, has excellent effects on anti-inflammation. However, its effect on microglia activation-induced inflammatory pain is not known yet. In the present study, a mouse BV2 microglia cell activation-mediated inflammatory model was developed with LPS induction, and a mouse inflammatory pain model was established with CFA injection. The inhibitory effect of muscone on microglia inflammatory activation was verified by measuring pro-inflammatory cytokines expression (interleukin-6, tumor necrosis factor-α, and interleukin-1β; IL-6, TNF-α and IL-1β). We found that muscone suppressed microglial activation-mediated inflammatory response through the NADPH oxidase 4 (NOX4)/janus kinase 2-signal transducer and activator of transcription 3 (JAK2-STAT3) pathway and pyrin-domain-containing 3 (NLRP3) inflammasome. Notably, muscone mitigated CFA-induced pain hypersensitivity and inflammation, as well as microglia cell activation in vivo. Furthermore, muscone inhibited the CFA-induced NOX4, p-JAK2/p-STAT3, and NLRP3 inflammasome expression in spinal cord of mice. In conclusion, this study uncovered that muscone relieved inflammatory pain by inhibiting microglial activation-mediated inflammatory response via abrogation of the NOX4/JAK2-STAT3 pathway and NLRP3 inflammasome. This finding of muscone is promising for treating inflammatory pain.

Keywords:  Inflammatory pain; JAK2-STAT3 pathway; Microglial activation; Muscone; NLRP3; NOX4

DOI:  https://doi.org/10.1016/j.intimp.2020.106355
Sci Rep. 2020 Mar 12. 10(1): 4570

Identification of highly potent and selective inhibitor, TIPTP, of the p22phox-Rubicon axis as a therapeutic agent for rheumatoid arthritis.

Ye-Ram Kim, Jae-Sung Kim, Su-Jin Gu, Sungsin Jo, Sojin Kim, Sun Young Kim, Daeun Lee, Kiseok Jang, Hyunah Choo, Tae-Hwan Kim, Jae U Jung, Sun-Joon Min, Chul-Su Yang.

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease linked to oxidative stress, which is associated with significant morbidity. The NADPH oxidase complex (NOX) produces reactive oxygen species (ROS) that are among the key markers for determining RA's pathophysiology. Therefore, understanding ROS-regulated molecular pathways and their interaction is necessary for developing novel therapeutic approaches for RA. Here, by combining mouse genetics and biochemistry with clinical tissue analysis, we reveal that in vivo Rubicon interacts with the p22phox subunit of NOX, which is necessary for increased ROS-mediated RA pathogenesis. Furthermore, we developed a series of new aryl propanamide derivatives consisting of tetrahydroindazole and thiadiazole as p22phox inhibitors and selected 2-(tetrahydroindazolyl)phenoxy-N-(thiadiazolyl)propanamide 2 (TIPTP, M.W. 437.44), which showed considerably improved potency, reaching an IC50 value up to 100-fold lower than an inhibitor that we previously synthesized reported N8 peptide-mimetic small molecule (blocking p22phox-Rubicon interaction). Notably, TIPTP treatment showed significant therapeutic effects a mouse model for RA. Furthermore, TIPTP had anti-inflammatory effects ex vivo in monocytes from healthy individuals and synovial fluid cells from RA patients. These findings may have clinical applications for the development of TIPTP as a small molecule inhibitor of the p22phox-Rubicon axis for the treatment of ROS-driven diseases such as RA.

DOI: https://doi.org/10.1038/s41598-020-61630-x