Carbon monoxide releasing molecule enhances coagulation and decreases fibrinolysis in canine plasma exposed to
            Crotalus viridis
            venom in vitro and in vivo

Tenascin-C (TN-C), an extracellular matrix (ECM) glycoprotein, is specifically induced upon tissue injury and infection and during septic conditions. Carbon monoxide (CO) gas is known to exert various anti-inflammatory effects in various inflammatory diseases. However, the mechanisms underlying the effect of CO on TN-C-mediated inflammation are unknown. In the present study, we found that treatment with LPS significantly enhanced TN-C expression in macrophages. CO gas, or treatment with the CO-donor compound, CORM-2, dramatically reduced LPS-induced expression of TN-C and proinflammatory cytokines while significantly increased the expression of IL-10. Treatment with TN-C siRNA significantly suppressed the effects of LPS on proinflammatory cytokines production. TN-C siRNA did not affect the CORM-2-dependent increase of IL-10 expression. In cells transfected with IL-10 siRNA, CORM-2 had no effect on the LPS-induced expression of TN-C and its downstream cytokines. These data suggest that IL-10 mediates the inhibitory effect of CO on TN-C and the downstream production of proinflammatory cytokines. Additionally, administration of CORM-2 dramatically reduced LPS-induced TN-C and proinflammatory cytokines production while expression of IL-10 was significantly increased. In conclusion, CO regulated IL-10 expression and thus inhibited TN-C-mediated inflammationin vitroandin vivo.

Download Full-text

Bioconversion of Carbon Monoxide to Formate Using Artificially Designed Carbon Monoxide:Formate Oxidoreductase in Hyperthermophilic Archaea

10.21203/rs.3.rs-132617/v1 ◽

2021 ◽

Author(s):

Jae Kyu Lim ◽

Ji-In Yang ◽

Yun Jae Kim ◽

Yeong-Jun Park ◽

Yong Hwan Kim

Keyword(s):

Carbon Monoxide ◽

Electron Transfer ◽

Fusion Protein ◽

Direct Electron Transfer ◽

Co2 Reduction ◽

Hyperthermophilic Archaea ◽

Production Activity ◽

Formate Production

Abstract Ferredoxin-dependent metabolic engineering of electron transfer circuits has been developed to enhance redox efficiency in the field of synthetic biology, e.g., for hydrogen production and for reduction of flavoproteins or NAD(P)+. Here, we present the bioconversion of carbon monoxide (CO) gas to formate via a synthetic CO:formate oxidoreductase (CFOR), designed as an enzyme complex for direct electron transfer between noninteracting CO dehydrogenase and formate dehydrogenase using an electron-transferring Fe-S fusion protein. The CFOR-introduced Thermococcus onnurineus mutant strains showed CO-dependent formate production in vivo and in vitro. The formate production rate from purified CFOR complex and specific formate productivity from the bioreactor were 348 ± 34 μmol/mg/min and 90.2 ± 20.4 mmol/g-cells/h, respectively. The CO-dependent CO2 reduction/formate production activity of synthetic CFOR was confirmed, indicating that direct electron transfer between two unrelated dehydrogenases was feasible via mediation of the FeS-FeS fusion protein.

Download Full-text

Ruthenium, Not Carbon Monoxide, Inhibits the Procoagulant Activity of Atheris, Echis, and Pseudonaja Venoms

International Journal of Molecular Sciences ◽

10.3390/ijms21082970 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2970 ◽

Cited By ~ 4

Author(s):

Vance G. Nielsen

Keyword(s):

Carbon Monoxide ◽

Phospholipase A2 ◽

Procoagulant Activity ◽

Inhibitory Effects ◽

Snake Venoms ◽

Radical Intermediate ◽

Experimental Systems ◽

Phospholipase A2 Activity

The demonstration that carbon monoxide releasing molecules (CORMs) affect experimental systems by the release of carbon monoxide, and not via the interaction of the inactivated CORM, has been an accepted paradigm for decades. However, it has recently been documented that a radical intermediate formed during carbon monoxide release from ruthenium (Ru)-based CORM (CORM-2) interacts with histidine and can inactivate bee phospholipase A2 activity. Using a thrombelastographic based paradigm to assess procoagulant activity in human plasma, this study tested the hypothesis that a Ru-based radical and not carbon monoxide was responsible for CORM-2 mediated inhibition of Atheris, Echis, and Pseudonaja species snake venoms. Assessment of the inhibitory effects of ruthenium chloride (RuCl3) on snake venom activity was also determined. CORM-2 mediated inhibition of the three venoms was found to be independent of carbon monoxide release, as the presence of histidine-rich albumin abrogated CORM-2 inhibition. Exposure to RuCl3 had little effect on Atheris venom activity, but Echis and Pseudonaja venom had procoagulant activity significantly reduced. In conclusion, a Ru-based radical and ion inhibited procoagulant snake venoms, not carbon monoxide. These data continue to add to our mechanistic understanding of how Ru-based molecules can modulate hemotoxic venoms, and these results can serve as a rationale to focus on perhaps other, complementary compounds containing Ru as antivenom agents in vitro and, ultimately, in vivo.

Download Full-text

A neuroglobin-based high-affinity ligand trap reverses carbon monoxide–induced mitochondrial poisoning

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.010593 ◽

2020 ◽

Vol 295 (19) ◽

pp. 6357-6371 ◽

Cited By ~ 2

Author(s):

Jason J. Rose ◽

Kaitlin A. Bocian ◽

Qinzi Xu ◽

Ling Wang ◽

Anthony W. DeMartino ◽

...

Keyword(s):

Carbon Monoxide ◽

Mitochondrial Respiration ◽

Oxygen Delivery ◽

Oxygen Electrode ◽

Heart Tissue ◽

Tissue Respiration ◽

Co Poisoning ◽

Affinity Ligand

Carbon monoxide (CO) remains the most common cause of human poisoning. The consequences of CO poisoning include cardiac dysfunction, brain injury, and death. CO causes toxicity by binding to hemoglobin and by inhibiting mitochondrial cytochrome c oxidase (CcO), thereby decreasing oxygen delivery and inhibiting oxidative phosphorylation. We have recently developed a CO antidote based on human neuroglobin (Ngb-H64Q-CCC). This molecule enhances clearance of CO from red blood cells in vitro and in vivo. Herein, we tested whether Ngb-H64Q-CCC can also scavenge CO from CcO and attenuate CO-induced inhibition of mitochondrial respiration. Heart tissue from mice exposed to 3% CO exhibited a 42 ± 19% reduction in tissue respiration rate and a 33 ± 38% reduction in CcO activity compared with unexposed mice. Intravenous infusion of Ngb-H64Q-CCC restored respiration rates to that of control mice correlating with higher electron transport chain CcO activity in Ngb-H64Q-CCC–treated compared with PBS-treated, CO-poisoned mice. Further, using a Clark-type oxygen electrode, we measured isolated rat liver mitochondrial respiration in the presence and absence of saturating solutions of CO (160 μm) and nitric oxide (100 μm). Both CO and NO inhibited respiration, and treatment with Ngb-H64Q-CCC (100 and 50 μm, respectively) significantly reversed this inhibition. These results suggest that Ngb-H64Q-CCC mitigates CO toxicity by scavenging CO from carboxyhemoglobin, improving systemic oxygen delivery and reversing the inhibitory effects of CO on mitochondria. We conclude that Ngb-H64Q-CCC or other CO scavengers demonstrate potential as antidotes that reverse the clinical and molecular effects of CO poisoning.

Download Full-text

Carbon monoxide in the treatment of sepsis

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00311.2015 ◽

2015 ◽

Vol 309 (12) ◽

pp. L1387-L1393 ◽

Cited By ~ 32

Author(s):

Kiichi Nakahira ◽

Augustine M. K. Choi

Keyword(s):

Carbon Monoxide ◽

Defense Mechanism ◽

Inflammatory Responses ◽

The Body ◽

High Concentration ◽

Novel Therapy ◽

Autophagy Induction ◽

Host Defense Mechanism

Carbon monoxide (CO), a low-molecular-weight gas, is endogenously produced in the body as a product of heme degradation catalyzed by heme oxygenase (HO) enzymes. As the beneficial roles of HO system have been elucidated in vitro and in vivo, CO itself has also been reported as a potent cytoprotective molecule. Whereas CO represents a toxic inhalation hazard at high concentration, low-dose exogenous CO treatment (∼250–500 parts per million) demonstrates protective functions including but not limited to the anti-inflammatory and antiapoptotic effects in preclinical models of human diseases. Of note, CO exposure confers protection in animal models of sepsis by inhibiting inflammatory responses and also enhancing bacterial phagocytosis in leukocytes. These unique functions of CO including both dampening inflammation and promoting host defense mechanism are mediated by multiple pathways such as autophagy induction or biosynthesis of specialized proresolving lipid mediators. We suggest that CO gas may represent a novel therapy for patients with sepsis.

Download Full-text

Exogenous Carbon Monoxide Produces Rapid Antidepressant- and Anxiolytic-Like Effects

Frontiers in Pharmacology ◽

10.3389/fphar.2021.757417 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yixiao Luo ◽

Rafi Ullah ◽

Jinfeng Wang ◽

Yuru Du ◽

Shihao Huang ◽

...

Keyword(s):

Carbon Monoxide ◽

Cell Injury ◽

Dorsal Hippocampus ◽

Protective Effects ◽

Neuroprotective Effects ◽

Gas Treatment ◽

Promising Therapeutic Target ◽

Co Gas

Carbon monoxide (CO), a byproduct of heme catalyzed by heme oxygenase (HO), has been reported to exert antioxidant and anti-inflammatory actions, and to produce significant neuroprotective effects. The potential effects of CO and even HO on depressive-like behaviors are still poorly understood. Utilizing several approaches including adeno-associated virus (AAV)-mediated overexpression of HO-1, systemic CO-releasing molecules (CO-RMs), CO-rich saline or CO gas treatment procedures in combination with hydrogen peroxide (H2O2)-induced PC12 cell injury model, and lipopolysaccharide (LPS)-induced depression mouse model, the present study aimed to investigate the potential antidepressant- and anxiolytic-like effects of endogenous and exogenous CO administration in vivo and in vitro. The results of in vitro experiments showed that both CO-RM-3 and CO-RM-A1 pretreatment blocked H2O2-induced cellular injuries by increasing cell survival and decreasing cell apoptosis and necrosis. Similar to the effects of CO-RM-3 and CO-RM-A1 pretreatment, AAV-mediated HO-1 overexpression in the dorsal hippocampus produced significant antidepressant-like activities in mice under normal conditions. Further investigation showed that the CO gas treatment significantly blocked LPS-induced depressive- and anxiety-like behaviors in mice. Taken together, our results suggest that the activation of HO-1 and/or exogenous CO administration produces protective effects and exerts antidepressant- and anxiolytic-like effects. These data uncover a novel function of the HO-1/CO system that appears to be a promising therapeutic target for the treatment of depression and anxiety.

Download Full-text

A Novel H2O2 Generator for Tumor Chemotherapy-Enhanced CO Gas Therapy

Frontiers in Oncology ◽

10.3389/fonc.2021.738567 ◽

2021 ◽

Vol 11 ◽

Author(s):

Yang Li ◽

Zeming Liu ◽

Weng Zeng ◽

Ziqi Wang ◽

Chunping Liu ◽

...

Keyword(s):

Carbon Monoxide ◽

Controlled Release ◽

Tumor Microenvironment ◽

Tumor Site ◽

Co Poisoning ◽

Chemotherapy Agent ◽

Manganese Carbonyl ◽

Co Gas

Carbon monoxide (CO) gas therapy is a promising cancer treatment. However, gas delivery to the tumor site remains problematic. Proper tunable control of CO release in tumors is crucial to increasing the efficiency of CO treatment and reducing the risk of CO poisoning. To overcome such challenges, we designed ZCM, a novel stable nanotechnology delivery system comprising manganese carbonyl (MnCO) combined with anticancer drug camptothecin (CPT) loaded onto a zeolitic imidazole framework-8 (ZIF-8). After intravenous injection, ZCM gradually accumulates in cancerous tissues, decomposing in the acidic tumor microenvironment, releasing CPT and MnCO. CPT acts as a chemotherapy agent destroying tumors and producing copious H2O2. MnCO can react with the H2O2 to generate CO, powerfully damaging the tumor. Both in vitro and in vivo experiments indicate that the ZCM system is both safe and has excellent tumor inhibition properties. ZCM is a novel system for CO controlled release, with significant potential to improve future cancer therapy.

Download Full-text

Thrombogenic Properties of Surface Factor; Evidence for an Anti-Surface Factor Activity in Canine Plasma.

Thrombosis and Haemostasis ◽

10.1055/s-0038-1655573 ◽

1964 ◽

Vol 12 (01) ◽

pp. 035-048 ◽

Cited By ~ 3

Author(s):

Sotirios G. Iatridis ◽

John H. Ferguson ◽

Panayotis G. Iatridis ◽

Ronald Mauldin

Keyword(s):

Mode Of Action ◽

Clotting Factor ◽

In Vitro Tests ◽

Inhibitory Mechanism ◽

Factor Activity ◽

Activation Product ◽

Canine Plasma ◽

Surface Factor

SummaryPurified surface factor (SF or activation product) was prepared from canine plasma and its effect was studied on 22 dogs. It was shown that intravenous injection of SF increased the coagulability of the circulating blood (in vitro tests) and induced thrombosis in areas of vascular stasis (in vivo assay). The hypercoagulability was associated with the injected SF preparation and with no other known clotting factor increase. A second injection performed 61 minutes after the first was less effective. The survival period of injected SF seemed to be relatively short. This might be attributable, in part, to an inhibitory mechanism acting against SF (anti-SF) in plasma. Experimental evidence of the existence of such an inhibitor and its “progressive” mode of action is provided. Anti-SF has a variable titer normally, but appears to increase after the intravenous SF injection in dogs.

Download Full-text

PPARγ Regulates the Anti-Inflammatory Effects of Carbon Monoxide on Macrophages: A Gene Profiling Study.

Blood ◽

10.1182/blood.v104.11.3445.3445 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3445-3445

Author(s):

Martin Bilban ◽

Sherrie L. Otterbein ◽

Emeka Ifedigbo ◽

Keiji Enjyoji ◽

Anny Usheva ◽

...

Keyword(s):

Carbon Monoxide ◽

Inflammatory Response ◽

Acute Phase Proteins ◽

Expression Patterns ◽

Endotoxic Shock ◽

Gene Profiling ◽

In Vivo Experiments ◽

Anti Inflammatory

Abstract Carbon monoxide (CO) at low concentrations has generated recent interest due to its ability to modulate the inflammatory response associated with chronic graft rejection, vascular injury and septic shock. Both in vivo and in vitro CO can inhibit the expression of pro-inflammatory genes such as TNFα in macrophages while simultaneously increasing the expression of the anti-inflammatory cytokine IL-10. The mechanisms by which this occurs are still unclear. To better understand the mechanisms underlying the effects of CO, we employed the Affymetrix GeneChip technology to evaluate the time-dependent expression patterns of >12,000 genes in macrophages stimulated with bacterial endotoxin (LPS) in the presence or absence of a low concentration of CO previously demonstrated to evoke an anti-inflammatory response. We were particularly interested whether CO would, by itself, modulate in a specific manner the expression of proteins that might explain the anti-inflammatory effects observed following subsequent administration of endotoxin. RAW 264.7 murine macrophages were grown to 75% confluency and then exposed to CO (250 ppm) for 3 hr prior to administration of LPS (10 ng/ml). At 0, 15, 30, 60, 120 and 240 min thereafter, total RNA was isolated by standard methods and the RNA was then labeled and hybridized to U74Av2 GeneChips. Of >12,000 genes assessed, 116 of 270 that were LPS-responsive were affected by CO treatment. CO inhibited the majority of LPS-induced pro-inflammatory cytokines and acute phase proteins including expression of early growth response-1 (Egr-1), a transcription factor that serves as a central intermediary regulating many genes. Egr-1 was nearly completely inhibited by CO as was Egr-1-dependent expression of tissue factor (TF) and PAI-1. Treatment of cells with CO alone led to a rapid early increase in PPARγ, the expression of which was essential for the anti-inflammatory effects of CO. Inhibition of PPARγ using the selective chemical inhibitor GW9662 reversed the CO inhibitory effects on LPS-induced Egr-1 and TF expression. Correlative in vivo experiments in mice showed that CO pre-treatment blocked endotoxin-induced Egr-1 expression and decreased markers of lung inflammmation the effects of which were also lost with inhibition of PPARγ. Our analyses of gene expression patterns has led to the first molecular understanding of how treatment with CO, in this case by inducing PPARγ, blocks the pro-inflammatory response. These experiments provide novel insights into the mechanisms and pathophysiology of endotoxic shock and identify cellular targets by which CO mediates these cytoprotective effects.

Download Full-text