In vivo effects of hemin and nitric oxide donors on parameters of heme metabolism in rat liver and serum

V N Karazin National University Series “Biology” ◽

10.26565/2075-5457-2018-31-2 ◽

2018 ◽

Keyword(s):

Blood Serum ◽

Heme Oxygenase ◽

Tbars Level ◽

No Donors ◽

Heme Synthesis ◽

Heme Degradation ◽

Free Heme ◽

In Vivo Effects ◽

Hemin Chloride

In vivo effects of hemin chloride (15 mg/kg body weight) and donors of nitrogen monoxide (NO) – sodium nitroprusside (SNP, 1 mg/kg) and substrate of NO-synthase L-arginine (L-Arg, 600 mg/kg) on the activity of key enzymes of heme synthesis (5-aminolevulinate synthase, ALAS) and heme degradation (heme oxygenase, HO), on the free heme level in liver and on the content of heme in blood serum of rats were studied. NO donors were administered alone or 30 min before hemin chloride injection. The level of free heme in liver was estimated by the ratio of holoenzyme and total tryptophan 2,3-dioxygenase (TDO) activities. Two hours after hemin chloride administration a significant increase in the level of heme-containing products and lipid peroxidation products (TBARS) was found in blood serum. These changes were accompanied by decrease in ALAS activity and by increase in holoenzyme activity and heme saturation of TDO, which was the result of free heme accumulation in liver. 24 hrs after administration of hemin chloride the content of heme in serum returned to normal level, while level of TBARS remained elevated. 24 hrs after hemin action a significant increase in the activities of HO and ALAS was observed in liver, while the degree of TDO heme saturation decreased, indicating the prevalence of heme degradation over its synthesis. Both NO donors did not affect the accumulation of heme in serum and liver first hours after hemin action. However, the specific features of SNP and L-Arg effects on the key enzyme of heme synthesis in liver and the TBARS level in serum were revealed. L-Arg, unlike SNP, prevented the accumulation of TBARS in serum, but did not prevent a decrease in ALAS activity 2 hrs after hemin chloride injection. The treatment by SNP itself caused an increase in TBARS level in serum, an increase in TDO activity and a decrease in ALAS activity in liver 2 hrs after action. Heme content in serum positively correlated with holoenzyme activity and heme saturation of TDO in liver. The pretreatment with NO donors did not affect the increase in HO activity, however, it blocked the induction of ALAS, a decrease in holoenzyme activity and heme saturation of TDO 24 hrs after the administration of hemin chloride. Thus, both SNP and Arg prevented a decrease in free heme level in liver, which might be due to heme nitrosylation in the presence of NO donors and, as a result, its slower degradation in the heme oxygenase reaction.

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Nitroglycerin as a Modulator of Hemoglobin Function.

Blood ◽

10.1182/blood.v108.11.1577.1577 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1577-1577

Author(s):

Borys Hrinczenko

Keyword(s):

Oxygen Affinity ◽

In Vitro Study ◽

Blood Substitutes ◽

Tissue Blood Flow ◽

Hepatic Microcirculation ◽

No Donors ◽

Biotransformation Products ◽

In Vivo Effects

Recent evidence suggests that oxygen acts as an allosteric modulator of nitric oxide (NO) binding to hemoglobin. This may have important physiological implications. Those effects, thought by some to be mediated by nitrosylation of the conserved cysteine β93 residues (S-nitrosohemoglobin; SNO-Hb), may also have therapeutic relevance in sickle cell disease and potentially be a useful strategy in controlling vasoactive effects of hemoglobin-based blood substitutes. Nitroglycerin (NTG), the classic vasodilator drug, acts in part through metabolism to NO. Normal (AA) and sickle (SS) blood suspensions were treated with nitroglycerin from 0.03 to 300 μM for periods up to 3.25 hours. Such treatment significantly lowered the oxygen affinity (increases P50, PO2 at which hemoglobin is half-saturated with O2) of both AA and SS red cells. The decrease in oxygen affinity is greater when oxygenated cells are treated than deoxygenated cells, with changes in P50 as great as 7.4 mm Hg in the AA cells and 9.9 mm Hg in the SS cells. Those shifts occur within 15 minutes, before significant increases in methemoglobin levels. This is opposite to what has been seen with other NO donors (2-(N,N-diethylamino)-diazenolate-2-oxide, S-nitrosocysteine, and sodium trioxodinitrate), where increased oxygen affinity occurs. Other classic NO donors also yielded significant methemoglobin formation within 15 minutes. In vivo effects of NTG infusion up to 1 μg/kg/min fail to demonstrate any change in P50 or methemoglobin in study subjects with normal AA erythrocytes. (Shigihara A. Masui1994;43:222) In addition, NTG has been shown to enhance oxygen release from erythrocytes without significantly increasing tissue blood flow in the hepatic microcirculation in a rat model. (Kosaka H, et al. Nat Med1997;3:456) Furthermore, a study in a dog model demonstrated NTG-induced enhanced oxygen unloading in the ischemic microcirculation. (Bin JP, et al. Circulation2006;113:2502) Elevated tandem erythrocyte S-nitrosothiol content, reflecting mainly SNO-Hb, was also noted. However, SNO-Hb increases oxygen affinity (Bonaventura C, et al. J Biol Chem1999;274:24742) and cannot explain the increased oxygen unloading. The effect of NTG on erythrocytes in our in vitro study model mimics in vivo results in the microcirculation (as opposed to the macrovascular circulation) and demonstrates that the reported enhanced oxygen unloading is due to a decrease in oxygen affinity of hemoglobin. Other biotransformation products of NTG may account for these effects as opposed to SNO-Hb. This study demonstrates that selective NO donors (i.e., NTG) may target erythrocytes for enhanced R (oxyhemoglobin) to T (deoxyhemoglobin) transformations specifically in the microcirculation and thereby improve tissue oxygenation.

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G-quadruplexes sequester free heme in living cells

10.1101/652297 ◽

2019 ◽

Author(s):

Lucas T. Gray ◽

Emilia Puig Lombardi ◽

Daniela Verga ◽

Corinne Guetta ◽

Alain Nicolas ◽

...

Keyword(s):

Iron Homeostasis ◽

Heme Degradation ◽

Expression Of Genes ◽

G Quadruplex ◽

Free Heme ◽

Key Enzyme ◽

Cultured Human Cells ◽

Fold Induction

AbstractHeme is an essential cofactor for many enzymes, but free heme is toxic and its levels are tightly regulated. G-quadruplexes bind heme avidlyin vitro, raising the possibility that they may sequester hemein vivo. If so, then treatment that displaces heme from quadruplexes is predicted to induce expression of genes involved in iron and heme homeostasis. Here we show that PhenDC3, a G-quadruplex ligand structurally unrelated to heme, displaces quadruplex-bound hemein vitroand alters transcription in cultured human cells, up-regulating genes that support heme degradation and iron homeostasis, and most strikingly causing a 30-fold induction of heme oxidase 1, the key enzyme in heme degradation. We propose that G-quadruplexes sequester heme to protect cells from the pathophysiological consequences of free heme. This identifies a new function for G-quadruplexes and a new mechanism for protection of cells from heme.

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Using ultrasound to define the time point of intrauterine growth retardation in a mouse model of heme oxygenase-1 deficiency†

Biology of Reproduction ◽

10.1093/biolre/ioaa057 ◽

2020 ◽

Vol 103 (1) ◽

pp. 126-134 ◽

Cited By ~ 2

Author(s):

Nicole Meyer ◽

Stefanie Langwisch ◽

Markus Scharm ◽

Ana Claudia Zenclussen

Keyword(s):

Heme Oxygenase ◽

Fetal Growth ◽

Molecular Mechanisms ◽

Maternal Blood ◽

Heme Oxygenase 1 ◽

Intrauterine Growth ◽

Flow Parameters ◽

Free Heme ◽

The Impact

Abstract The enzyme heme oxygenase-1 (HO-1), encoded by the HMOX1 gene, mediates heme catabolism by cleaving free heme. We have previously revealed the importance of HO-1 in pregnancy. Here, we determined the impact of maternal or paternal HO-1 deficiency on fetal growth and placental parameters throughout gestation. We mated Hmox1-sufficient (WT), partial (HET)-, or total (KO)-deficient BALB/c female mice with Hmox1-WT or -KO BALB/c males and performed ultrasound analysis to monitor placental and fetal growth. Doppler measurements were used to determine maternal blood flow parameters. Offspring weights and feto-placental indices (FPI) were also determined. We found a significantly increased number of underdeveloped fetuses at gd10 in HET females that were mated with WT males compared with WT × WT pairings. At the same gestational age, underdeveloped placentas could be detected in HET females mated with KO males. Many fetuses from the KO × KO combination died in utero between gd12 and gd14. At gd14, abnormal placental parameters were found in surviving fetuses, which had significant reduced weights. Moreover, only 3.11% female and 5.33% male KO pups resulted from 10 HET × HET breeding pairs over 1 year. Our results show that HO-1 from both maternal and paternal origins is important for proper placental and fetal growth. Placental growth restriction and occurrence of abortions in mice that were partially or totally deficient in HO-1 were recorded in vivo from gd10 onwards. Future studies will focus on elucidating the cellular and molecular mechanisms behind these observations.

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Effect of reduced glutathione on the indexes of oxidative stress and heme metabolism in liver and blood of rats under hemin chloride injection in vivo

V N Karazin National University Series “Biology” ◽

10.26565/2075-5457-2019-33-1 ◽

2019 ◽

Keyword(s):

Oxidative Stress ◽

Body Weight ◽

Blood Plasma ◽

Reduced Glutathione ◽

Water Soluble ◽

Lipid Hydroperoxides ◽

Free Heme ◽

Radical Processes ◽

Hemin Chloride

Heme (iron-protoporphyrin IX) is involved in various cellular functions. The release of heme under hemolysis or under the damage of intracellular hemeproteins leads to its accumulation in tissues and, as a result, to the activation of free radical processes. Reduced glutathione (GSH) functions as an endogenous water-soluble antioxidant and a regulator of cells redox status, but its effect on the development of oxidative stress under hemin action in mammals remains not investigated. The aim of this work was to study the effect of hemin chloride on some hemeproteins activity and a number of prooxidant-antioxidant status indexes in rat liver and blood under GSH level modulation in vivo. White male rats weighing 170–280 g were taken for investigation. Hemin chloride and GSH were injected intraperitoneally. Blood plasma, homogenate, and postmitochondrial fraction of liver were the objects of study. Hemin chloride injection (50 mg/kg body weight) caused the increase in heme-containing products level in blood and free heme level in liver of rats, which was accompanied by the activation of free radical processes in these tissues. The accumulation of free heme in liver was proved by an increase in tryptophan 2,3-dioxygenase (TDO) holoenzyme activity and heme saturation. The pretreatment by GSH (500 mg/kg body weight) 0.5 h before hemin chloride injection normalized GSH content, but did not prevent heme accumulation, the decrease in triglycerides level and the increase in lipid hydroperoxides content in rat blood plasma under hemin action. In liver, GSH injection prevented the increase in lipid hydroperoxides and protein carbonyl derivatives concentration as well as in TDO holoenzyme activity, and decreased the degree of TDO heme saturation. All these changes occurred under GSH content increase in liver. Catalase activity in liver did not differ from the control values after hemin chloride injection as well as after glutathione and hemin coadministration. The analysis of relationship between parameters studied in this work revealed the strong positive correlation between GSH content in plasma and liver (r=0.85; p<0.001), which was consistent with literature data on the significant role of liver in supplying other tissues with reduced glutathione. A negative correlation was found between lipid peroxidation products and triglycerides content in plasma (r=–0.52; p<0.05), which indicated the participation of triglycerides unsaturated fatty acids as substrates in the peroxidation processes under hemin action. No significant correlation between GSH and hydroperoxides content, as well as between GSH and heme-containing products levels in blood plasma was revealed. Thus, the water-soluble antioxidant glutathione was not effective enough to prevent damage of lipid components in blood under hemin chloride action in the selected dose. In the liver, on the contrary, GSH injection prevented heme accumulation and oxidative stress development under hemin action, which was obviously associated with an increase in the GSH content in this organ.

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Protein aggregation as a cellular response to oxidative stress induced by heme and iron

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1608928113 ◽

2016 ◽

Vol 113 (47) ◽

pp. E7474-E7482 ◽

Cited By ~ 41

Author(s):

Luiz R. C. Vasconcellos ◽

Fabianno F. Dutra ◽

Mariana S. Siqueira ◽

Heitor A. Paula-Neto ◽

Jennifer Dahan ◽

...

Keyword(s):

Cellular Response ◽

Innate Immune Responses ◽

Free Iron ◽

Ubiquitinated Proteins ◽

Heme Degradation ◽

Expression Of Genes ◽

Proinflammatory Molecule ◽

Free Heme ◽

Necessary And Sufficient

Hemolytic diseases include a variety of conditions with diverse etiologies in which red blood cells are destroyed and large amounts of hemeproteins are released. Heme has been described as a potent proinflammatory molecule that is able to induce multiple innate immune responses, such as those triggered by TLR4 and the NLRP3 inflammasome, as well as necroptosis in macrophages. The mechanisms by which eukaryotic cells respond to the toxic effects induced by heme to maintain homeostasis are not fully understood, however. Here we describe a previously uncharacterized cellular response induced by heme: the formation of p62/SQTM1 aggregates containing ubiquitinated proteins in structures known as aggresome-like induced structures (ALIS). This action is part of a response driven by the transcription factor NRF2 to the excessive generation of reactive oxygen species induced by heme that results in the expression of genes involved in antioxidant responses, including p62/SQTM1. Furthermore, we show that heme degradation by HO-1 is required for ALIS formation, and that the free iron released on heme degradation is necessary and sufficient to induce ALIS. Moreover, ferritin, a key protein in iron metabolism, prevents excessive ALIS formation. Finally, in vivo, hemolysis promotes an increase in ALIS formation in target tissues. Our data unravel a poorly understood aspect of the cellular responses induced by heme that can be explored to better understand the effects of free heme and free iron during hemolytic diseases such as sickle cell disease, dengue fever, malaria, and sepsis.

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837 Absence of in-vivo effects of dental instruments on pacemaker function

EP Europace ◽

10.1016/s1099-5129(05)80575-6 ◽

2005 ◽

Vol 7 ◽

pp. 195-195

Keyword(s):

Dental Instruments ◽

Pacemaker Function ◽

In Vivo Effects

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In vivo effects of TGF-β1 in lung surfactant regulation, lung meachanics amd structure

Pneumologie ◽

10.1055/s-0034-1376812 ◽

2014 ◽

Vol 68 (06) ◽

Author(s):

E Lopez-Rodriguez ◽

C Boden ◽

S Knippenberg ◽

A Pascual ◽

J Perez-Gil ◽

...

Keyword(s):

Lung Surfactant ◽

In Vivo Effects ◽

Tgf Β1

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Evaluation of Sacroiliac Wedge Rotation to Increase Acetabular Ventroversion

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0038-1632486 ◽

1999 ◽

Vol 12 (04) ◽

pp. 173-177 ◽

Cited By ~ 2

Author(s):

R. L. Aper ◽

M. D. Brown ◽

M. G. Conzemius

Keyword(s):

Hip Joint ◽

Hip Dysplasia ◽

Clinical Effect ◽

Pelvic Osteotomy ◽

Alternative Method ◽

Angular Measurements ◽

Si Joint ◽

In Vivo Effects ◽

Canine Hip Dysplasia

SummaryTreatment of canine hip dysplasia (CHD) via triple pelvic osteotomy (TPO) is widely accepted as the treatment that best preserves the existing hip joint. TPO, however, has several important disadvantages. In an effort to avoid some of the difficulties associated with TPO an alternative method of creating acetabular ventroversion (AW) was sought. The purpose of this study was to explore the effects of placement of a wedge in the sacroiliac (SI) joint on A W and to compare this to the effect of TPO on A W . On one hemipelvis a 30° pelvic osteotomy plate was used for TPO. The contralateral hemipelvis had a 28° SI wedge inserted into the SI joint. Pre- and postsurgical radiographs of each pelvis were taken and the angular measurements were recorded. On average, the 28° SI wedge resulted in 20.9° of A W, the 30° canine pelvic osteotomy plate resulted in 24.9° A W . Significant differences were not found (p >0.05) between the two techniques. Sacroiliac wedge rotation effectively creates A W and has several theoretical advantages when compared to TPO. The in vivo effects of sacroiliac wedge rotation should be studied in order to evaluate the clinical effect of the technique.Sacroiliac wedge rotation was tested as an alternative method to increase the angle of acetabular ventroversion. This technique effectively rotated the acetabulum and has several theoretical advantages when compared to triple pelvic osteotomy.

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