bims-noxint 2022-04-24 papers

bims-noxint

Biomed News

on NADPH oxidases in tumorigenesis

Issue of 2022–04–24
eleven papers selected by
Laia Caja Puigsubira, Uppsala University

HDAC1 expression is positively correlated with NADPH oxidase 4-mediated oxidative stress in a mouse model of traumatic brain injury.
The Notch pathway attenuates burn-induced acute lung injury in rats by repressing reactive oxygen species.
Euphorbia supina Extracts Block NADPH Oxidase-Mediated, Ceramide-Induced Apoptosis Initiated by Diesel Particulate Matter.
NOX activation in reactive astrocytes regulates astrocytic LCN2 expression and neurodegeneration.
Setanaxib (GKT137831) Ameliorates Doxorubicin-Induced Cardiotoxicity by Inhibiting the NOX1/NOX4/Reactive Oxygen Species/MAPK Pathway.
Novel potential NOX2 inhibitors, Dudleya brittonii water extract and polygalatenoside A inhibit intracellular ROS generation and growth of melanoma.
Cellular signals converge at the NOX2-SHP-2 axis to induce reductive carboxylation in cancer cells.
Impact of klotho on the expression of SRGAP2a in podocytes in diabetic nephropathy.
Hydroxysafflor Yellow A Blocks HIF-1α Induction of NOX2 and Protects ZO-1 Protein in Cerebral Microvascular Endothelium.
GABA and Fermented Curcuma longa L. Extract Enriched with GABA Ameliorate Obesity through Nox4-IRE1α Sulfonation-RIDD-SIRT1 Decay Axis in High-Fat Diet-Induced Obese Mice.
Exercise Affects the Formation and Recovery of Alcoholic Liver Disease through the IL-6-p47phox Oxidative-Stress Axis.

J Neurophysiol. 2022 Apr 20.

HDAC1 expression is positively correlated with NADPH oxidase 4-mediated oxidative stress in a mouse model of traumatic brain injury.

Yiming Chen, Yiming Lu, Siyi Xu, Min Liu, Wei Chen, Yanfei Zhang, Liang Wei, Chunlong Zhong.

  Accumulating evidence has demonstrated that histone deacetylase 1 (HDAC1) expression is statistically correlated with the severity of traumatic brain injury (TBI). However, the specific role of HDAC1 in the occurrence and development of TBI remains unclear. The Lateral Fluid Percussion Injury (LFPI) was used to conduct TBI mouse model in C57BL/6J and C57BL/6J-Hdac1em1cyagen mice. Western blot and qRT-PCR were performed to estimate the expression of HADC1 and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in brain tissues. Modified Neurological Severity Score (mNSS) and brain water content were performed to detect the neurological deficit. Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were used to detect the oxidative stress. Oxidative stresses, HDAC1 and NOX4 expression were upregulated in the lesioned cortices tissues after TBI. HDAC1 protein expression was positively correlated with the NOX4 in TBI mouse. Hdac1 knockout attenuated brain edema and neurological dysfunction caused by TBI in mice. Hdac1 knockout inhibited the expressions of NOX4 induced by TBI and attenuated TBI induced oxidative stress. HDAC1 expression is positively correlated with to NOX4-mediated oxidative stress in a TBI mouse model.

Keywords:  HDAC1; NOX4; Oxidative stress; Traumatic brain injury (TBI)

DOI:  https://doi.org/10.1152/jn.00049.2022
Burns Trauma. 2022 ;10 tkac008

The Notch pathway attenuates burn-induced acute lung injury in rats by repressing reactive oxygen species.

Weixia Cai, Kuo Shen, Peng Ji, Yanhui Jia, Shichao Han, Wanfu Zhang, Xiaolong Hu, Xuekang Yang, Juntao Han, Dahai Hu.

   Background: Acute lung injury (ALI) is a common complication following severe burns. The underlying mechanisms of ALI are incompletely understood; thus, available treatments are not sufficient to repair the lung tissue after ALI.
Methods: To investigate the relationship between the Notch pathway and burn-induced lung injury, we established a rat burn injury model by scalding and verified lung injury via lung injury evaluations, including hematoxylin and eosin (H&E) staining, lung injury scoring, bronchoalveolar lavage fluid and wet/dry ratio analyses, myeloperoxidase immunohistochemical staining and reactive oxygen species (ROS) accumulation analysis. To explore whether burn injury affects Notch1 expression, we detected the expression of Notch1 and Hes1 after burn injury. Then, we extracted pulmonary microvascular endothelial cells (PMVECs) and conducted Notch pathway inhibition and activation experiments, via a γ-secretase inhibitor (GSI) and OP9-DLL1 coculture, respectively, to verify the regulatory effect of the Notch pathway on ROS accumulation and apoptosis in burn-serum-stimulated PMVECs. To investigate the regulatory effect of the Notch pathway on ROS accumulation, we detected the expression of oxidative-stress-related molecules such as superoxide dismutase, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) 2, NOX4 and cleaved caspase-3. NOX4-specific small interfering RNA (siRNA) and the inhibitor GKT137831 were used to verify the regulatory effect of the Notch pathway on ROS via NOX4.
Results: We successfully established a burn model and revealed that lung injury, excessive ROS accumulation and an inflammatory response occurred. Notch1 detection showed that the expression of Notch1 was significantly increased after burn injury. In PMVECs challenged with burn serum, ROS and cell death were elevated. Moreover, when the Notch pathway was suppressed by GSI, ROS and cell apoptosis levels were significantly increased. Conversely, these parameters were reduced when the Notch pathway was activated by OP9-DLL1. Mechanistically, the inhibition of NOX4 by siRNA and GKT137831 showed that the Notch pathway reduced ROS production and cell apoptosis by downregulating the expression of NOX4 in PMVECs.
Conclusions: The Notch pathway reduced ROS production and apoptosis by downregulating the expression of NOX4 in burn-stimulated PMVECs. The Notch-NOX4 pathway may be a novel therapeutic target to treat burn-induced ALI.

Keywords:  Acute lung injury; Burn; Nicotinamide adenine dinucleotide phosphate oxidase 4; Notch pathway; Pulmonary microvascular endothelial cells; Reactive oxygen species

DOI:  https://doi.org/10.1093/burnst/tkac008
Pharmaceuticals (Basel). 2022 Mar 31. pii: 431. [Epub ahead of print]15(4):

Euphorbia supina Extracts Block NADPH Oxidase-Mediated, Ceramide-Induced Apoptosis Initiated by Diesel Particulate Matter.

Kyong-Oh Shin, Sungeun Kim, Bokyung Kim, Hye-Yoon Park, Eunhee Jung, Garyun Kim, Donghee Kim, Hwang Eui Cho, Yoshikazu Uchida, Kyungho Park.

  Air pollutants contribute to the development of diseases such as asthma, chronic obstructive pulmonary disease (COPD), pulmonary cancer, cardiovascular problems, and some skin diseases. We recently found that a major air pollutant, diesel particulate matter (DPM), induces apoptosis in human keratinocytes by increasing a proapoptotic lipid mediator, ceramide. DPM activates nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX), which stimulates sphingomyelinase, leading to an increased conversion of sphingomyelin to ceramide. Interestingly, we characterized that although NOX is a reactive oxygen species (ROS) generator, the activation of sphingomyelinases by NOX is an ROS-independent mechanism. A Korean weed, prostrate spurge Euphorbia supina Rafin (ESR), has been used for centuries as a folk medicine to treat bronchitis, hepatitis, hemorrhage, and skin inflammation. Flavonoids, terpenes and tannins are enriched in ESR, and although ESR has proven antioxidative activity, its biological activities are largely unknown. Here, we investigate whether and how ESR protects keratinocytes against DPM-mediated apoptosis. We found that ESR-extracts (ESR-Ex) protect keratinocytes from DPM-induced apoptosis by inhibiting NOX activation in keratinocytes in response to DPM. We also demonstrated that ESR-Ex suppresses NOX activation via a blockage of the aryl hydrocarbon receptor (AhR) activation-mediated transcription of neutrophil cytosolic factor 1 (NCF1)/p47phox, a subunit of NOX. Our study reveals previously uncharacterized biological activity of ESR-Ex; i.e., its inhibition of Ahr and NOX activation. Thus, because the inhibition of NOX has already been developed to treat NOX-mediated diseases, including various types of cardiovascular diseases and cancers, initiated by air pollutants and because AhR activation contributes to the development of chronic inflammatory diseases, our study provides further advantages for the medical use of ESR.

Keywords:  Euphorbia supina Rafin; apoptosis; ceramide; diesel particulate matters; nicotinamide adenine dinucleotide phosphate oxidase; sphingomyelinase

DOI:  https://doi.org/10.3390/ph15040431
Cell Death Dis. 2022 Apr 19. 13(4): 371

NOX activation in reactive astrocytes regulates astrocytic LCN2 expression and neurodegeneration.

Ruijia Liu, Jun Wang, Yang Chen, Jenelle M Collier, Okan Capuk, Shijie Jin, Ming Sun, Sujan K Mondal, Theresa L Whiteside, Donna B Stolz, Yongjie Yang, Gulnaz Begum.

Reactive astrocytes (RA) secrete lipocalin-2 (LCN2) glycoprotein that regulates diverse cellular processes including cell death/survival, inflammation, iron delivery and cell differentiation. Elevated levels of LCN2 are considered as a biomarker of brain injury, however, the underlying regulatory mechanisms of its expression and release are not well understood. In this study, we investigated the role of astrocytic Na+/H+ exchanger 1 (NHE1) in regulating reactive astrocyte LCN2 secretion and neurodegeneration after stroke. Astrocyte specific deletion of Nhe1 in Gfap-CreER+/-;Nhe1f/f mice reduced astrogliosis and astrocytic LCN2 and GFAP expression, which was associated with reduced loss of NeuN+ and GRP78+ neurons in stroke brains. In vitro ischemia in astrocyte cultures triggered a significant increase of secreted LCN2 in astrocytic exosomes, which caused neuronal cell death and neurodegeneration. Inhibition of NHE1 activity during in vitro ischemia with its potent inhibitor HOE642 significantly reduced astrocytic LCN2+ exosome secretion. In elucidating the cellular mechanisms, we found that stroke triggered activation of NADPH oxidase (NOX)-NF-κB signaling and ROS-mediated LCN2 expression. Inhibition of astrocytic NHE1 activity attenuated NOX signaling and LCN2-mediated neuronal apoptosis and neurite degeneration. Our findings demonstrate for the first time that RA use NOX signaling to stimulate LCN2 expression and secretion. Blocking astrocytic NHE1 activity is beneficial to reduce LCN2-mediated neurotoxicity after stroke.

DOI: https://doi.org/10.1038/s41419-022-04831-8
Front Pharmacol. 2022 ;13 823975

Setanaxib (GKT137831) Ameliorates Doxorubicin-Induced Cardiotoxicity by Inhibiting the NOX1/NOX4/Reactive Oxygen Species/MAPK Pathway.

Hui Zheng, Nannan Xu, Zihao Zhang, Fen Wang, Jie Xiao, Xiaoping Ji.

  Background: Doxorubicin (DOX)-induced cardiotoxicity is a highly concerning issue, and the mechanism by which DOX induces cardiotoxicity is likely to be multifactorial. NADPH oxidase (NOX) is associated with DOX-induced cardiotoxicity. Setanaxib (GKT137831), a preferential direct inhibitor of NOX1 and NOX4, can delay or prevent the progression of many cardiovascular disorders by inhibiting reactive oxygen species (ROS) generation. In this study, we investigated the role of GKT137831 in ameliorating DOX-induced cardiotoxicity and the potential mechanisms of its action. Methods and Results: The mice model of cardiotoxicity induced by DOX was established, and GKT137831 treatment was performed at the same time. Neonatal rat cardiomyocytes (NRCMs) were treated with DOX or GKT137831 for in vitro experiments. We found that DOX administration impaired cardiac function in vivo, reflected by decreased left ventricular ejection fraction (LVEF) and fractional shortening (FS%). DOX also impaired the viability of NRCMs in vitro. In addition, DOX increased the levels of NOX1 and NOX4 expression and ROS production and the cardiomyocyte apoptosis rate, both in vivo and in vitro. GKT137831 improved cardiac function, as indicated by the increased LVEF and FS%. In vitro, GKT137831 improved NRCM viability. It also decreased ROS production and the cardiomyocyte apoptosis rate. Apoptotic indices, such as cleaved PARP (c-PARP), cleaved caspase 3 (CC3) and BAX expression levels, were decreased, and the antiapoptotic index of Bcl-2 expression was increased. DOX markedly activated phosphorylated JNK, ERK and p38 proteins in NRCMs. Specific inhibitors of JNK (SP600125), ERK (PD98059) or p38 (SB203580) inhibited DOX-induced apoptosis of NRCMs. GKT137831 pretreatment inhibited excessive DOX-induced MAPK pathway activation. Conclusion: This study revealed that GKT137831 can alleviate DOX-induced cardiomyocyte apoptosis by inhibiting NOX1/4-driven ROS production. The upregulation of MAPK pathway induced by NOX1/4-derived ROS production may be the potential mechanism of GKT137831 action. GKT137831 may be a potential drug candidate to ameliorate DOX-induced cardiotoxicity.

Keywords:  GKT137831; NADPH oxidase; apoptosis; cardiotoxicity; doxorubicin

DOI:  https://doi.org/10.3389/fphar.2022.823975
Biomed Pharmacother. 2022 Apr 14. pii: S0753-3322(22)00356-0. [Epub ahead of print]150 112967

Novel potential NOX2 inhibitors, Dudleya brittonii water extract and polygalatenoside A inhibit intracellular ROS generation and growth of melanoma.

Hyungkuen Kim, Eunmi Hwang, Byung-Chul Park, Sung-Jo Kim.

  Reactive oxygen species (ROS) are key regulators of the proliferation, metastasis, and drug resistance of melanoma, which accounts for 60% of skin cancer deaths. In a previous study, we developed Dudleya brittonii water extract (DBWE) with antioxidant activity, but the mechanism of action and bioactive substances of DBWE have not been fully identified. This study showed altered NADPH oxidase 2 (NOX2) expression and selective inhibition of cytosolic ROS but not mitochondrial ROS in B16-F10 melanoma cells, suggesting the NOX2 inhibitory potential of DBWE. In addition, DBWE inhibited mitochondrial activity, lipid metabolism, and cell cycle in B16-F10 cells. The anti-melanoma effect of DBWE was abrogated by the addition of ROS, and there was no significant change in the melanogenesis pathway. Polygalatenoside A was identified as a candidate bioactive substance in the DBWE aqueous fraction through mass spectrometry, and the DBWE-like anti-melanoma effect was confirmed. These data suggest that DBWE and polygalatenoside A have the potential to prevent and treat melanoma.

Keywords:  Antioxidant; Dudleya brittonii; Melanoma; NADPH oxidase; Polygalatenoside; Reactive oxygen species

DOI:  https://doi.org/10.1016/j.biopha.2022.112967
Cell Chem Biol. 2022 Apr 13. pii: S2451-9456(22)00127-1. [Epub ahead of print]

Cellular signals converge at the NOX2-SHP-2 axis to induce reductive carboxylation in cancer cells.

Rukang Zhang, Dong Chen, Hao Fan, Rong Wu, Jiayi Tu, Freya Q Zhang, Mei Wang, Hong Zheng, Cheng-Kui Qu, Shannon E Elf, Brandon Faubert, Yu-Ying He, Marc B Bissonnette, Xue Gao, Ralph J DeBerardinis, Jing Chen.

  Environmental stresses, including hypoxia or detachment for anchorage independence, or attenuation of mitochondrial respiration through inhibition of electron transport chain induce reductive carboxylation in cells with an enhanced fraction of citrate arising through reductive metabolism of glutamine. This metabolic process contributes to redox homeostasis and sustains biosynthesis of lipids. Reductive carboxylation is often dependent on cytosolic isocitrate dehydrogenase 1 (IDH1). However, whether diverse cellular signals induce reductive carboxylation differentially or through a common signaling converging node remains unclear. We found that induction of reductive carboxylation commonly requires enhanced tyrosine phosphorylation and activation of IDH1, which, surprisingly, is achieved by attenuation of a cytosolic protein tyrosine phosphatase, Src homology region 2 domain-containing phosphatase-2 (SHP-2). Mechanistically, diverse signals induce reductive carboxylation by converging at upregulation of NADPH oxidase 2, leading to elevated cytosolic reactive oxygen species that consequently inhibit SHP-2. Together, our work elucidates the signaling basis underlying reductive carboxylation in cancer cells.

Keywords:  NADPH oxidase 2 (NOX2); Src homology region 2 domain-containing phosphatase-2 (SHP-2); cytosolic reactive oxygen species (ROS); isocitrate dehydrogenase 1 (IDH1); reductive carboxylation; tyrosine phosphorylation

DOI:  https://doi.org/10.1016/j.chembiol.2022.03.010
BMC Nephrol. 2022 Apr 18. 23(1): 151

Impact of klotho on the expression of SRGAP2a in podocytes in diabetic nephropathy.

Donghua Jin, Miao Jia, Yuxian Xie, Lihua Lin, Hong Qiu, Guoyuan Lu.

   BACKGROUND: Diabetic nephropathy (DN) is the major cause of kidney failure, and glomerular podocytes play critical roles in the pathogenesis of DN by maintaining the glomerular structure and filtration barrier. Klotho and Slit-Robo GTP activating protein 2a (SRGAP2a) have been indicated to play protective roles in reducing kidney injury, but whether there is an internal relationship between these two factors is unclear.
METHODS: In this study, we cultured differentiated rat podocytes in vitro and measured the SRGAP2a expressions by immunofluorescence staining, quantitative real-time PCR (qRT-PCR) and western blotting, after siRNA-mediated transforming growth factor β1 (TGF-β1) silencing, TGF-β1 overexpression and in the presence of a reactive oxygen species (ROS) inhibitor. And we detected the expressions of SRGAP2a, small mother against decapentaplegic (Smad)2/3, phosphorylated-Smad2/3 (p-Smad2/3), Smad7, and NAD(P)H oxidase 4 (NOX4), ROS levels and podocyte cytoskeletal remodelling under high glucose (HG) and exogenous klotho conditions. In addition, we performed haematoxylin-eosin (HE) staining and immunohistochemistry with diabetic rat models to confirm the in vitro results.
RESULTS: The results indicated that SRGAP2a expression was significantly upregulated under siRNA-mediated TGF-β1 silencing conditions or after adding a ROS inhibitor, but significantly downregulated with TGF-β1 overexpression, in the presence of HG. The supplementation of exogenous klotho under HG conditions significantly increased the SRGAP2a expression, remodelled the actin cytoskeleton and altered the expressions of Smad2/3, p-Smad2/3, Smad7 and NOX4 and reduced the ROS generation in podocytes. Moreover, klotho administration protected kidney injury in DN rats.
CONCLUSIONS: This study indicated that klotho may modulate the expression of SRGAP2a by regulating the ROS and TGF-β1 signalling pathways and provided theoretical support for klotho protein as a novel therapeutic strategy for treating DN patients.

Keywords:  Diabetic nephropathy; Klotho; Podocyte; SRGAP2a

DOI:  https://doi.org/10.1186/s12882-022-02765-z
Antioxidants (Basel). 2022 Apr 07. pii: 728. [Epub ahead of print]11(4):

Hydroxysafflor Yellow A Blocks HIF-1α Induction of NOX2 and Protects ZO-1 Protein in Cerebral Microvascular Endothelium.

Yi Li, Xiao-Tian Liu, Pei-Lin Zhang, Yu-Chen Li, Meng-Ru Sun, Yi-Tao Wang, Sheng-Peng Wang, Hua Yang, Bao-Lin Liu, Mei Wang, Wen Gao, Ping Li.

  Zonula occludens-1 (ZO-1) is a tight junction protein in the cerebrovascular endothelium, responsible for blood-brain barrier function. Hydroxysafflor yellow A (HSYA) is a major ingredient of safflower (Carthamus tinctorius L.) with antioxidative activity. This study investigated whether HSYA protected ZO-1 by targeting ROS-generating NADPH oxidases (NOXs). HSYA administration reduced cerebral vascular leakage with ZO-1 protection in mice after photothrombotic stroke, largely due to suppression of ROS-associated inflammation. In LPS-stimulated brain microvascular endothelial cells, HSYA increased the ratio of NAD+/NADH to restore Sirt1 induction, which bound to Von Hippel-Lindau to promote HIF-1αdegradation. NOX2 was the predominant isoform of NOXs in endothelial cells and HIF-1α transcriptionally upregulated p47phox and Nox2 subunits for the assembly of the NOX2 complex, but the signaling cascades were blocked by HSYA via HIF-1α inactivation. When oxidate stress impaired ZO-1 protein, HSYA attenuated carbonyl modification and prevented ZO-1 protein from 20S proteasomal degradation, eventually protecting endothelial integrity. In microvascular ZO-1 deficient mice, we further confirmed that HSYA protected cerebrovascular integrity and attenuated ischemic injury in a manner that was dependent on ZO-1 protection. HSYA blocked HIF-1α/NOX2 signaling cascades to protect ZO-1 stability, suggestive of a potential therapeutic strategy against ischemic brain injury.

Keywords:  HIF-1α; NOX2; ZO-1; cerebral microvascular endothelium; hydroxysafflor yellow A

DOI:  https://doi.org/10.3390/antiox11040728
Nutrients. 2022 Apr 18. pii: 1680. [Epub ahead of print]14(8):

GABA and Fermented Curcuma longa L. Extract Enriched with GABA Ameliorate Obesity through Nox4-IRE1α Sulfonation-RIDD-SIRT1 Decay Axis in High-Fat Diet-Induced Obese Mice.

Hwa-Young Lee, Geum-Hwa Lee, The-Hiep Hoang, Yu-Mi Kim, Gi-Hyun Jang, Chang-Hwan Seok, Yun-Geum-Sang Gwak, Junghyun Lim, Junghyun Kim, Han-Jung Chae.

  Gamma-aminobutyric acid (GABA) is a natural amino acid with antioxidant activity and is often considered to have therapeutic potential against obesity. Obesity has long been linked to ROS and ER stress, but the effect of GABA on the ROS-associated ER stress axis has not been thoroughly explored. Thus, in this study, the effect of GABA and fermented Curcuma longa L. extract enriched with GABA (FCLL-GABA) on the ROS-related ER stress axis and inositol-requiring transmembrane kinase/endoribonuclease 1α (IRE1α) sulfonation were examined with the HFD model to determine the underlying anti-obesity mechanism. Here, GABA and FCLL-GABA supplementations significantly inhibited the weight gain in HFD fed mice. The GABA and FCLL-GABA supplementation lowered the expressions of adipogenic transcription factors such as PPAR-γ, C/EBPα, FAS, and SREBP-1c in white adipose tissue (WAT) and liver from HFD-fed mice. The enhanced hyper-nutrient dysmetabolism-based NADPH oxidase (Nox) 4 and the resultant IRE1α sulfonation-RIDD-SIRT1 decay under HFD conditions were controlled with GABA and FCLL-GABA. Notably, GABA and FCLL-GABA administration significantly increased AMPK and sirtuin 1 (SIRT1) levels in WAT of HFD-fed mice. These significant observations indicate that ER-localized Nox4-induced IRE1α sulfonation results in the decay of SIRT1 as a novel mechanism behind the positive implications of GABA on obesity. Moreover, the investigation lays a firm foundation for the development of FCLL-GABA as a functional ingredient.

Keywords:  IRE1α; NADPH oxidase; SIRT1; decay; obesity

DOI:  https://doi.org/10.3390/nu14081680
Cells. 2022 Apr 12. pii: 1305. [Epub ahead of print]11(8):

Exercise Affects the Formation and Recovery of Alcoholic Liver Disease through the IL-6-p47phox Oxidative-Stress Axis.

Wei Cui, Xiang Li, Weiyue Xue, Huiting Wei, Gang Zhou, Ye Qiu, Di Cui.

  (1) Background: To explore the effect of exercise on the formation and recovery of alcoholic liver disease (ALD) and whether the IL-6-p47phox oxidative-stress axis is involved in that process. (2) Methods: Firstly, 23 six-week-old male C57BL/6J mice were randomly divided into the Con group, ALD group, ALD + NOXI group, ALD + Ex group, and ALD + Ex + NOXI group. The Liber-DeCarli alcoholic liquid diet was used for 6 weeks to establish the ALD mice model, and the Con group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet, and exercise intervention was started after the ALD model establishment and lasted for another 6 weeks, with or without administration of the NOX inhibitor apocynin by intraperitoneal injection on every exercise training day. Secondly, 28 mice were randomly divided into the Sed group, Eth group, Eth + Ex group and Eth + Ex + NOXI group. The Sed group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet and exercise intervention was started synchronously combined with or without administration of intraperitoneal apocynin injections on every exercise training day for 5 weeks. After each individual experiment was accomplished, physiological assessment and biochemical analysis of blood and tissue samples were examined. (3) Results: The levels of TG in serum and IL-6 protein content in liver tissue in the ALD group were significantly increased compared to the Con group (p < 0.05); compared with ALD, p47phox expression in muscle was increased significantly in the ALD + NOXI group (p < 0.05), and TG in serum decreased in the ALD + Ex group (p < 0.05). TG in serum, AST/ALT ratio, and IL-6 content in both liver and muscle decreased (p < 0.05) in the ALD + Ex + NOXI group with MDA in muscle significantly increased (p < 0.01). The AST/ALT ratio, TG in serum, SOD in liver, and p47phox in both liver and muscle in the ALD + Ex + NOXI group were significantly decreased compared with the ALD + NOXI group (p < 0.01). Compared with the ALD + Ex group, the liver index and HDL-C levels in serum were decreased (p < 0.05) in the ALD + Ex + NOXI group. The degree of hepatocyte steatosis and inflammatory infiltration were ameliorated after exercise intervention. In the Eth group, the relative epididymal fat content, HDL-C level, and AST/ALT ratio were significantly decreased, and TG and gp91phox in liver were significantly higher than in the Sed group (p < 0.05, p < 0.01). Compared with the Eth group, the AST/ALT ratio, MDA in the liver, and NOX4 and p47phox protein expression in the liver were significantly increased, and body weight decreased significantly in the Eth + Ex group (p < 0.05, p < 0.01), as did TG in the liver and MDA in muscle. In the th + Ex + NOXI group, gp91phox expression in the liver and body weight were significantly decreased (p < 0.05, p < 0.01). In the Eth + Ex + NOXI group, the ratio of AST/ALT and MDA in muscle were increased when compared with the Eth + Ex group, and the protein expression of gp91phox and p47phox were much lower (p < 0.01). (4) Conclusions: 6 weeks of exercise intervention during the recovery phase of ALD ameliorates hepatocyte damage and dyslipidemia through the IL-6-p47phox oxidative-stress axis, and applying a NOX inhibitor in combination could optimize this. However, drinking alcohol during exercise exacerbates dyslipidemia and oxidative stress, with hepatocyte IL-6-p47phox downregulated.

Keywords:  NADPH oxidase (NOX); alcoholic liver disease (ALD); exercise; hepatocellular; inflammation; interleukin-6 (IL-6); oxidative stress

DOI:  https://doi.org/10.3390/cells11081305