Activation of Rho/Rho kinase signaling pathway by reactive oxygen species in rat aorta

Evidence indicates that both the Rho/Rho kinase signaling pathway and reactive oxygen species (ROS) such as superoxide and H2O2 are involved in the pathogenesis of hypertension. This study aimed to determine whether ROS-induced vascular contraction is mediated through activation of Rho/Rho kinase. Rat aortic rings (endothelium denuded) were isolated and placed in organ chambers for measurement of isometric force development. ROS were generated by a xanthine (X)-xanthine oxidase (XO) mixture. The antioxidants tempol (3 mM) and catalase (1,200 U/ml) or the XO inhibitor allopurinol (400 μM) significantly reduced X/XO-induced contraction. A Rho kinase inhibitor, (+)-( R)- trans-4-(1-aminoethyl- N-4-pyridil)cyclohexanecarboxamide dihydrochloride (Y-27632), decreased the contraction in a concentration-dependent manner; however, the Ca2+-independent protein kinase C inhibitor rottlerin did not have an effect on X/XO-induced contraction. Phosphorylation of the myosin light chain phosphatase target subunit (MYPT1) was increased by ROS, and preincubation with Y-27632 blocked this increased phosphorylation. Western blotting for cytosolic and membrane-bound fractions of Rho showed that Rho was increased in the membrane fraction by ROS, suggesting activation of Rho. These observations demonstrate that ROS-induced Ca2+ sensitization is through activation of Rho and a subsequent increase in Rho kinase activity but not Ca2+-independent PKC.

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Baicalin Induces Apoptosis and Inhibits Migration/ Invasion of Breast Cancer Cell MDA-MB-231 Through Reactive Oxygen Species-Mediated Activation of p38/c-Jun N-Terminal Kinase Signaling Pathway

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2018.1838 ◽

2018 ◽

Vol 8 (7) ◽

pp. 1022-1030 ◽

Cited By ~ 2

Author(s):

Huayong Zhang ◽

Xinfeng Huang ◽

Qiaomin Li ◽

Beilong Zhong ◽

Qiang Han

Keyword(s):

Breast Cancer ◽

Reactive Oxygen Species ◽

Signaling Pathway ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Reactive Oxygen ◽

Oxygen Species ◽

Kinase Signaling ◽

Kinase Signaling Pathway

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Erigeron annuus (L.) Pers. Extract Inhibits Reactive Oxygen Species (ROS) Production and Fat Accumulation in 3T3-L1 Cells by Activating an AMP-Dependent Kinase Signaling Pathway

Antioxidants ◽

10.3390/antiox8050139 ◽

2019 ◽

Vol 8 (5) ◽

pp. 139 ◽

Cited By ~ 5

Author(s):

Yoon-Hee Choi ◽

Ok-Hwan Lee ◽

Yulong Zheng ◽

Il-Jun Kang

Keyword(s):

Reactive Oxygen Species ◽

Signaling Pathway ◽

Water Extract ◽

Reactive Oxygen ◽

Dependent Manner ◽

Ros Production ◽

Oxygen Species ◽

Fat Accumulation ◽

Ampk Signaling ◽

Erigeron Annuus

Obesity is one of the major public health problems in the world because it is implicated in metabolic syndromes, such as type 2 diabetes, hypertension, and cardiovascular diseases. The objective of this study was to investigate whether Erigeron annuus (L.) Pers. (EAP) extract suppresses reactive oxygen species (ROS) production and fat accumulation in 3T3-L1 cells by activating an AMP-dependent kinase (AMPK) signaling pathway. Our results showed that EAP water extract significantly inhibits ROS production, adipogenesis, and lipogenesis during differentiation of 3T3-L1 preadipocytes. In addition, EAP decreased mRNA and protein levels of proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα). Moreover, EAP suppressed mRNA expressions of fatty acid synthase (FAS), lipoprotein lipase (LPL), adipocyte protein 2 (aP2) in a dose-dependent manner. Whereas, EAP upregulated adiponectin expression, phosphorylation levels of AMPK and carnitine palmitoyltransferase 1 (CPT-1) protein level during differentiation of 3T3-L1 preadipocytes. These results suggest that EAP water extract can exert ROS-linked anti-obesity effect through the mechanism that might involve inhibition of ROS production, adipogenesis and lipogenesis via an activating AMPK signaling pathway.

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Iron overload induced by ferric ammonium citrate triggers reactive oxygen species-mediated apoptosis via both extrinsic and intrinsic pathways in human hepatic cells

Human & Experimental Toxicology ◽

10.1177/0960327115597312 ◽

2015 ◽

Vol 35 (6) ◽

pp. 598-607 ◽

Cited By ~ 13

Author(s):

S-W Li ◽

C-M Liu ◽

J Guo ◽

AM Marcondes ◽

J Deeg ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Iron Overload ◽

Cell Viability ◽

Signaling Pathway ◽

Reactive Oxygen ◽

Ammonium Citrate ◽

Ferric Ammonium Citrate ◽

Dependent Manner ◽

Oxygen Species ◽

Ferric Ammonium

Background: Hepatic iron overload is common in patients with myelodysplastic syndromes undergoing hematopoietic cell transplantation (HCT) and may predispose to peri- and post-HCT toxicity. To better understand the mechanisms of iron overload-induced liver injury, we examined the effects of iron overload induced by ferric ammonium citrate (FAC) on oxidative stress and apoptosis signaling pathway in human hepatic cell line HH4. Methods and Results: Hepatic HH4 cells were exposed to FAC to force iron uptake, and cellular responses were determined. Incubation with 5 mM FAC resulted in increased intracellular iron content in a time-dependent manner. High concentration of FAC impaired cell viability and increased level of reactive oxygen species (ROS), and addition of antioxidant reagent such as glutathione or N-acetylcysteine dramatically reduced FAC-induced intracellular ROS generation. FAC overload significantly increased the phosphorylation of inhibitor of κB-α, p38 mitogen-activated protein kinase (MAPK), and nuclear factor κ light chain enhancer of activated B cells (NF-κB) p65 and promoted the nuclear translocation of NF-κB p65. Knockdown of Fas and Bid expression by small interfering RNA in iron-treated HH4 cells resulted in restoration of cell viability. Conclusions: We reported that FAC treatment is capable of inducing both extrinsic death receptor and intrinsic mitochondrial signaling pathway-mediated HH4 cells apoptosis through ROS-activated p38 MAPK and NF-κB pathways.

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Abstract 522: Contribution of Reactive Oxygen Species and Rho Kinase in Sphingosine-1-phosphate Mediated Afferent Arteriolar Vasoconstriction of Ischemia-Reperfusion Rats

Hypertension ◽

10.1161/hyp.64.suppl_1.522 ◽

2014 ◽

Vol 64 (suppl_1) ◽

Author(s):

Zhengrong Guan ◽

Edward W. Inscho

Keyword(s):

Reactive Oxygen Species ◽

Kinase Inhibitor ◽

Ischemia Reperfusion ◽

Rho Kinase ◽

Reactive Oxygen ◽

Kidney Injury ◽

Oxygen Species ◽

Sphingosine 1 Phosphate ◽

Arteriolar Tone

Our recent studies indicate that sphingosine-1-phosphate (S1P) is a potent vasoconstrictor of afferent arterioles (AA) and is enhanced in renal ischemia-reperfusion (IR) injury. We hypothesized that renal IR-induced increases in reactive oxygen species (ROS) and activation of the rho kinase pathway contribute to the enhanced S1P sensitivity of AA in IR rats. IR injury was induced by bilateral renal artery occlusion for 60 min followed by 24 hrs reperfusion. Renal function was assessed by plasma creatinine (Cr) or glomerular filtration rate (GFR). The AA response to S1P was assessed using the in vitro blood-perfused juxtamedullary nephron preparation with and without superfusion with the ROS scavenger, Tempol, or the rho kinase inhibitor, Y27623. Plasma Cr was markedly increased in IR (3.86±0.20 vs. 0.93±0.07 mg/L in sham, n=5-7, P < 0.05). GFR, calculated from the half time (T 1/2 ) of sinistrin clearance, was significantly reduced in IR rats. The T 1/2 averaged 166±28 min in IR vs. 25±4 min in sham rats corresponding to GFRs of 0.20±0.02 and 1.33±0.23 ml/min (P < 0.05, n=3 each), respectively. The basal AA diameter was significantly decreased in IR compared to sham rats (10.2±0.5 vs. 15.0±0.4 μm, P < 0.05, n=9 each). Superfusion of Tempol (10 -4 M) did not alter the basal AA diameter in sham but increased diameter in IR by 8±1% (P < 0.05, n=6 each). S1P superfusion evoked concentration-dependent AA vasoconstriction in both groups, however, Tempol significantly shifted the S1P response curve to the right in IR. Increasing concentrations of S1P (10 -10 -10 -5 M) reduced AA diameter to 98±2, 96±2, 98±2, 94±3, 81±9 and 59±5% of control in IR rats similar to the response in Tempol-treated sham rats (99±1, 99±2, 96±2, 88±5, 65±6 and 48±4%, respectively, P > 0.05). In contrast, superfusion of Y27632 (10 -5 M) markedly increased the basal AA diameter in both groups. AA diameter increased by 81±17% from 10.6±1.3 to 19.2±3.3 μm in IR vs. a 53±21% increase in sham rats (P > 0.05, n=3 each). Y27632 also significantly blunted S1P-mediated AA vasoconstriction similarly in both groups. These results indicate a significant contribution of ROS and rho kinase to both the elevated afferent arteriolar tone and the enhanced S1P sensitivity of AA in IR induced acute kidney injury.

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Emodin induced necroptosis and inhibited glycolysis in the renal cancer cells by enhancing ROS

10.21203/rs.3.rs-67745/v1 ◽

2020 ◽

Author(s):

Ke-jie Wang ◽

Xiang-yu Meng ◽

Jun-feng Chen ◽

Kai-yun Wang ◽

Cheng Zhou ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Flow Cytometry ◽

Cancer Cells ◽

Signaling Pathway ◽

Renal Cancer ◽

Reactive Oxygen ◽

Chinese Herbs ◽

Dependent Manner ◽

Oxygen Species ◽

Flow Cytometry Assay

Abstract Background: Renal cell carcinoma (RCC) is a tumor with unpredictable presentation and poor clinical outcome. RCC is always resistant to chemotherapy, radiation, and weakly sensitive to immunotherapeutic agents. Therefore, novel agents and approaches are urgently needed for the treatment of RCC. Emodin, an anthraquinone compound extracted from rhubarb and other traditional Chinese herbs, has been implicated in a wide variety of pharmacological effects, such as anti-inflammatory, antiviral, and antitumor activities. However，its role in RCC remains unkown. Methods: Flow cytometry assay and lactate dehydrogenase release assay were used to detect the cell death. Reactive oxygen species was tested by the dye MitoSox and DCFH-DA. Glucose-6-phosphate, pyruvate and ATP level were measured to evaluate the glycolysis process. Western blot was used to detect protein expression. Results: Emodin effectively killed renal cancer cells without significant toxicity to normal renal tubular epithelial cell. Flow cytometry assay with Annexin V-FITC and PI demonstrated that emodin induces necroptosis, but not apoptosis, in renal cancer cells. Meanwhile, the phosphorylation levels of RIP1 and MLKL, the key necroptosis-related proteins, were significantly increased. To explore how emodin inhibits kidney tumor growth, reactive oxygen species (ROS) levels were tested and the levels of ROS increased upon emodin treatment in a dose-dependent manner. Further studies demonstrated that emodin induced necroptosis through ROS-mediated activation of JNK signaling pathway, and also inhibited glycolysis by down-regulation GLUT1 by ROS-mediated inactivation of PI3K/AKT signaling pathway. Conclusion: These findings revealed the potential mechanisms by which emodin suppresses renal cancer cell growth, and will help develop novel therapeutic approaches for patients with JNK- or PI3K/AKT-dysregulated renal cancer.

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Actin polymerization contributes to enhanced pulmonary vasoconstrictor reactivity after chronic hypoxia

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00664.2017 ◽

2018 ◽

Vol 314 (5) ◽

pp. H1011-H1021 ◽

Cited By ~ 7

Author(s):

Laura Weise-Cross ◽

Michelle A. Sands ◽

Joshua R. Sheak ◽

Brad R. S. Broughton ◽

Jessica B. Snow ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Actin Polymerization ◽

Chronic Hypoxia ◽

Pulmonary Arteries ◽

Rho Kinase ◽

Reactive Oxygen ◽

Dependent Manner ◽

Oxygen Species ◽

Pulmonary Arterial ◽

Arterial Constriction

Chronic hypoxia (CH) augments basal and endothelin-1 (ET-1)-induced pulmonary vasoconstrictor reactivity through reactive oxygen species (ROS) generation and RhoA/Rho kinase (ROCK)-dependent myofilament Ca2+ sensitization. Because ROCK promotes actin polymerization and the actin cytoskeleton regulates smooth muscle tension, we hypothesized that actin polymerization is required for enhanced basal and ET-1-dependent vasoconstriction after CH. To test this hypothesis, both end points were monitored in pressurized, endothelium-disrupted pulmonary arteries (fourth-fifth order) from control and CH (4 wk at 0.5 atm) rats. The actin polymerization inhibitors cytochalasin and latrunculin attenuated both basal and ET-1-induced vasoconstriction only in CH vessels. To test whether CH directly alters the arterial actin profile, we measured filamentous actin (F-actin)-to-globular actin (G-actin) ratios by fluorescent labeling of F-actin and G-actin in fixed pulmonary arteries and actin sedimentation assays using homogenized pulmonary artery lysates. We observed no difference in actin polymerization between groups under baseline conditions, but ET-1 enhanced actin polymerization in pulmonary arteries from CH rats. This response was blunted by the ROS scavenger tiron, the ROCK inhibitor fasudil, and the mDia (RhoA effector) inhibitor small-molecule inhibitor of formin homology domain 2. Immunoblot analysis revealed an effect of CH to increase both phosphorylated (inactive) and total levels of the actin disassembly factor cofilin but not phosphorylated cofilin-to-total cofilin ratios. We conclude that actin polymerization contributes to increased basal pulmonary arterial constriction and ET-1-induced vasoconstrictor reactivity after CH in a ROS- and ROCK-dependent manner. Our results further suggest that enhanced ET-1-mediated actin polymerization after CH is dependent on mDia but independent of changes in the phosphorylated cofilin-to-total cofilin ratio. NEW & NOTEWORTHY This research is the first to demonstrate a role for actin polymerization in chronic hypoxia-induced basal pulmonary arterial constriction and enhanced agonist-induced vasoconstrictor activity. These results suggest that a reactive oxygen species-Rho kinase-actin polymerization signaling pathway mediates this response and may provide a mechanistic basis for the vasoconstrictor component of pulmonary hypertension.

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Reactive oxygen species rescue regeneration after silencing the MAPK–ERK signaling pathway in Schmidtea mediterranea

Scientific Reports ◽

10.1038/s41598-020-79588-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

V. Jaenen ◽

S. Fraguas ◽

K. Bijnens ◽

M. Heleven ◽

T. Artois ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Signaling Pathway ◽

Reactive Oxygen ◽

Light Therapy ◽

Erk Signaling ◽

Model Organisms ◽

Ros Production ◽

Oxygen Species ◽

Planarian Regeneration ◽

Egfr Signaling Pathway

AbstractDespite extensive research on molecular pathways controlling the process of regeneration in model organisms, little is known about the actual initiation signals necessary to induce regeneration. Recently, the activation of ERK signaling has been shown to be required to initiate regeneration in planarians. However, how ERK signaling is activated remains unknown. Reactive Oxygen Species (ROS) are well-known early signals necessary for regeneration in several models, including planarians. Still, the probable interplay between ROS and MAPK/ERK has not yet been described. Here, by interfering with major mediators (ROS, EGFR and MAPK/ERK), we were able to identify wound-induced ROS, and specifically H2O2, as upstream cues in the activation of regeneration. Our data demonstrate new relationships between regeneration-related ROS production and MAPK/ERK activation at the earliest regeneration stages, as well as the involvement of the EGFR-signaling pathway. Our results suggest that (1) ROS and/or H2O2 have the potential to rescue regeneration after MEK-inhibition, either by H2O2-treatment or light therapy, (2) ROS and/or H2O2 are required for the activation of MAPK/ERK signaling pathway, (3) the EGFR pathway can mediate ROS production and the activation of MAPK/ERK during planarian regeneration.

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Protective Effects of Gintonin on Reactive Oxygen Species-Induced HT22 Cell Damages: Involvement of LPA1 Receptor-BDNF-AKT Signaling Pathway

Molecules ◽

10.3390/molecules26144138 ◽

2021 ◽

Vol 26 (14) ◽

pp. 4138

Author(s):

Yeon-Jin Cho ◽

Sun-Hye Choi ◽

Ra-Mi Lee ◽

Han-Sung Cho ◽

Hyewhon Rhim ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Signaling Pathway ◽

Reactive Oxygen ◽

Akt Signaling ◽

Atp Depletion ◽

Oxygen Species ◽

Akt Signaling Pathway ◽

Neuroprotective Effects ◽

Lpa1 Receptor

Gintonin is a kind of ginseng-derived glycolipoprotein that acts as an exogenous LPA receptor ligand. Gintonin has in vitro and in vivo neuroprotective effects; however, little is known about the cellular mechanisms underlying the neuroprotection. In the present study, we aimed to clarify how gintonin attenuates iodoacetic acid (IAA)-induced oxidative stress. The mouse hippocampal cell line HT22 was used. Gintonin treatment significantly attenuated IAA-induced reactive oxygen species (ROS) overproduction, ATP depletion, and cell death. However, treatment with Ki16425, an LPA1/3 receptor antagonist, suppressed the neuroprotective effects of gintonin. Gintonin elicited [Ca2⁺]i transients in HT22 cells. Gintonin-mediated [Ca2⁺]i transients through the LPA1 receptor-PLC-IP3 signaling pathway were coupled to increase both the expression and release of BDNF. The released BDNF activated the TrkB receptor. Induction of TrkB phosphorylation was further linked to Akt activation. Phosphorylated Akt reduced IAA-induced oxidative stress and increased cell survival. Our results indicate that gintonin attenuated IAA-induced oxidative stress in neuronal cells by activating the LPA1 receptor-BDNF-TrkB-Akt signaling pathway. One of the gintonin-mediated neuroprotective effects may be achieved via anti-oxidative stress in nervous systems.

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