Increase In Methylglyoxal Research Articles

Compensatory mechanisms for methylglyoxal detoxification in experimental & clinical diabetes

ObjectivesThe deficit of Glyoxalase I (Glo1) and the subsequent increase in methylglyoxal (MG) has been reported to be one the five mechanisms by which hyperglycemia causes diabetic late complications. Aldo-keto reductases (AKR) have been shown to metabolize MG; however, the relative contribution of this superfamily to the detoxification of MG in vivo, particularly within the diabetic state, remains unknown. MethodsCRISPR/Cas9-mediated genome editing was used to generate a Glo1 knock-out (Glo1−/−) mouse line. Streptozotocin was then applied to investigate metabolic changes under hyperglycemic conditions. ResultsGlo1−/− mice were viable and showed no elevated MG or MG-H1 levels under hyperglycemic conditions. It was subsequently found that the enzymatic efficiency of various oxidoreductases in the liver and kidney towards MG were increased in the Glo1−/− mice. The functional relevance of this was supported by the altered distribution of alternative detoxification products. Furthermore, it was shown that MG-dependent AKR activity is a potentially clinical relevant pathway in human patients suffering from diabetes. ConclusionsThese data suggest that in the absence of GLO1, AKR can effectively compensate to prevent the accumulation of MG. The combination of metabolic, enzymatic, and genetic factors, therefore, may provide a better means of identifying patients who are at risk for the development of late complications caused by elevated levels of MG.

Methylglyoxal Disrupts Paranodal Axoglial Junctions via Calpain Activation.

Nodes of Ranvier and associated paranodal and juxtaparanodal domains along myelinated axons are essential for normal function of the peripheral and central nervous systems. Disruption of these domains as well as increases in the reactive carbonyl species methylglyoxal are implicated as a pathophysiology common to a wide variety of neurological diseases. Here, using an ex vivo nerve exposure model, we show that increasing methylglyoxal produces paranodal disruption, evidenced by disorganized immunostaining of axoglial cell-adhesion proteins, in both sciatic and optic nerves from wild-type mice. Consistent with previous studies showing that increase of methylglyoxal can alter intracellular calcium homeostasis, we found upregulated activity of the calcium-activated protease calpain in sciatic nerves after methylglyoxal exposure. Methylglyoxal exposure altered clusters of proteins that are known as calpain substrates: ezrin in Schwann cell microvilli at the perinodal area and zonula occludens 1 in Schwann cell autotypic junctions at paranodes. Finally, treatment with the calpain inhibitor calpeptin ameliorated methylglyoxal-evoked ezrin loss and paranodal disruption in both sciatic and optic nerves. Our findings strongly suggest that elevated methylglyoxal levels and subsequent calpain activation contribute to the disruption of specialized axoglial domains along myelinated nerve fibers in neurological diseases.

Exogenous glutathione attenuates lead-induced oxidative stress in wheat by improving antioxidant defense and physiological mechanisms

ABSTRACTThis study aims at investigate how exogenous glutathione (GSH, 1.0 mM) affects the oxidative stress and antioxidant defense in wheat seedlings under lead (Pb) stress [0.5 and 1.0 mM Pb(NO3)2]. Lead treatment decreased growth, leaf relative water content, and chlorophyll (chl) content whereas raised proline (Pro) level. Lead stress increased H2O2 content, generation rate, and membrane lipid peroxidation. Addition of Pb also disrupted antioxidant enzyme activities and status of endogenous ascorbate and GSH pool. The increase of methylglyoxal was evident under Pb stress. Glutathione supplementation under Pb stress increased antioxidant redox pool and augmented the activities of antioxidant enzymes, and decreased ROS production. Exogenous supplementation of GSH reverted the increase in the methylglyoxal level due to Pb stress due to increased activities of glyoxalase enzymes. Exogenous GSH also regulated Pro, well-maintained tissue water status and prevented chl degradation and increased plant growth and biomass. Abbreviations: AO- ascorbate oxidase; APX- ascorbate peroxidase; AsA- ascorbic acid (ascorbate); BSA- bovine serum albumin; CAT- catalase; CDNB-1-chloro-2, 4-dinitrobenzene; chl- chlorophyll; DHA- dehydroascorbate; DHAR- dehydroascorbate reductase; DTNB- 5,5′-dithio-bis (2-nitrobenzoic acid); EDTA- ethylenediaminetetraacetic acid; Gly I- glyoxalase I; Gly II- glyoxalase II; GR- glutathione reductase; GSH- reduced glutathione; GSSG- oxidized glutathione; GPX- glutathione peroxidase; GST- glutathione S-transferase; MDA- malondialdehyde; MDHA- monodehydroascorbate; MDHAR- monodehydroascorbate reductase; MG- methylglyoxal; NADPH- nicotinamide adenine dinucleotide phosphate; NBT- nitroblue tetrazolium chloride; Pb- lead; PEG- polyethylene glycol; Pro- proline, ROS- reactive oxygen species; RWC- relative water content; SLG- S-d-lactoylglutathione; SOD- superoxide dismutase; TBA- thiobarbituric acid; TCA- trichloroacetic acid.

English

Drought is one of the most important stress factors which adversely affect plants’ growth and productivity. Global climate change may make this situation more serious in the years ahead. Considering the long time span required for the generation of drought resistant genotypes in Rubber (Hevea brasiliensis) through conventional breeding, molecular interventions to engineer plants to have either drought responsive genes or genes expected to alter osmotic regulation would be very attractive. The glyoxalase pathway involving glyoxalase I and glyoxalase II enzymes is required for glutathione-based detoxification of methylglyoxal. In this study the effects of various abiotic stresses on the up-regulation of methylglyoxal levels and glyoxalase I activities in Hevea brasiliensis seedlings were investigated. Most of the stresses caused significant increase in methylglyoxal level and glyoxalase I activity, among which drought caused the highest induction of glx I followed by salinity, 2, 4-D, ABA, methylglyoxal, white light and CdCl2. The stress-induced increases in methylglyoxal and glyoxalase I activity found in the present study suggest an important role of glyoxalase I in conferring drought tolerance. The up-regulation of glyoxalase I under drought stress indicates its future utility in developing tolerance to drought stress in Hevea brasiliensis. In the present study, a partial cDNA sequence coding for glyoxalase I was amplified by PCR using specific primers. The 440 bp cDNA amplicon obtained was sequenced and subjected to online BLAST analysis. The sequence of Hevea brasiliensis glyoxalase I (GenBank Acc. No: GU598520) had six open reading frames. The ORF finder revealed the longest ORF of 336 bp. Glyoxalase I from Ricinus communis had the highest nucleotide sequence homology (90%) compared to the amplified gene. BLASTP analysis also showed high homology between the deduced protein sequence of the amplified gene and glyoxalases from other species. Our results suggest that the multi-stress inducibility of glyoxalase I in the present study may be due to the fact that it might protect the plants against MG that is formed under various stresses including drought and confers tolerance by increasing the GSH-based detoxification system and decreasing lipid peroxidation Key words: Glyoxalase I, Hevea brasiliensis, Methylglyoxal, drought tolerance, abiotic stress.

Differentiation and apoptosis induction by lovastatin and γ-tocotrienol in HL-60 cells via Ras/ERK/NF-κB and Ras/Akt/NF-κB signaling dependent down-regulation of glyoxalase 1 and HMG-CoA reductase

Glyoxalase 1 (GLO1) and HMG-CoA reductase (HMGCR) are highly expressed in most tumor cells and little in normal cells. In this study, treatment of HL-60 cells with lovastatin induced characteristic apoptosis in a dose-dependent manner. We demonstrated that lovastatin treatment inhibited Ras and Raf protein translocation to cell membrane and eliminated the phosphorylation of the downstream effectors Akt and ERK, and the subsequent NF-κB translocation into nucleus. Specific inhibitors and γ-tocotrienol confirmed the Ras/Raf/ERK/NF-κB/GLO1 and Ras/Akt/NF-κB/GLO1 pathways. Data revealed that lovastatin induced HL-60 cell death was attenuated by mevalonate treatment. We demonstrated also that γ-tocotrienol showed its apoptotic effect on the HL-60 cell through the same pathway. γ-Tocotrienol enhanced the apoptotic effect of lovastatin through the down-regulation of GLO1 and HMGCR resulting in an increase of methylglyoxal and a decrease of cholesterol and led to the apoptosis of HL-60 cells.Data also revealed that both lovastatin and gamma-tocotrienol induced significant HL-60 cell differentiation. These results suggest that both lovastatin and gamma-tocotrienol could induce differentiation and followed by apoptosis.

Fructose-induced stress signaling in the liver involves methylglyoxal

BackgroundFructose produces hepatic insulin resistance in humans and animals. We have proposed that the selective metabolism of fructose by the liver can, under conditions of elevated fructose delivery, inflict a metabolic insult that is localized to the hepatocyte. The present study was designed to identify potential cellular effectors of this insult.MethodsPrimary hepatocytes were incubated with 8 mM glucose and 0.12% inulin (G, n = 6) or 8 mM glucose, 0.12% inulin and 28 mU of inulinase (GF, n = 6) in the presence or absence of insulin for 0, 2, or 4 h.ResultsGF produced fructose concentrations of ~0.7 mM over the 4 h experiment. GF induced phosphorylation of MKK7 and JNK, phosphorylation of serine307 on IRS-1, and reduced tyrosine phosphorylation of IRS-1 and -2. GF increased ceramide levels and reactive oxygen species (ROS); however inhibitors of ceramide synthesis or ROS accumulation did not prevent GF-mediated changes in MKK7, JNK or IRS proteins. GF increased cellular methylglyoxal concentrations and a selective increase in methylglyoxal recapitulated the GF-induced changes in MKK7, JNK and IRS proteins.ConclusionsWe hypothesize that GF-mediated changes in stress signaling involve methylglyoxal in primary hepatocytes.

Methyl glyoxal increase apoptosis (CASPASE-3 expression) in pre-osteoblast MC3T3E1 cell line via SOD activity

Methyl glyoxal increase apoptosis (CASPASE-3 expression) in pre-osteoblast MC3T3E1 cell line via SOD activity

Transgenic potato overproducing l-ascorbic acid resisted an increase in methylglyoxal under salinity stress via maintaining higher reduced glutathione level and glyoxalase enzyme activity

Salt-tolerance was studied in transgenic potato. It was conferred by overexpression of ascorbate pathway enzyme (D-galacturonic acid reductase, GalUR). As genetic engineering of the GalUR gene in potato enhances its ascorbic acid content (L-AsA), and subsequently plants suffered minimal oxidative stress-induced damage, we now report on the comprehensive aptness of this engineering approach for enhanced salt tolerance in transgenic potato (Solanum tuberosum L. cv. Taedong Valley). Potatoes overexpressing GalUR grew and tuberized in continuous presence of 200 mM of NaCl. The transgenic plants maintained a higher reduced to oxidized glutathione (GSH:GSSG) ratio together with enhanced activity of glutathione dependent antioxidative and glyoxalase enzymes under salinity stress. The transgenics resisted an increase in methylglyoxal that increased radically in untransformed control plants under salinity stress. This is the first report of genetic engineering of ascorbate pathway gene in maintaining higher level of GSH homeostasis along with higher glyoxalase activity inhibiting the accumulation in methylglyoxal (a potent cytotoxic compound) under salt stress. These results suggested the engineering of ascorbate pathway enzymes as a major step towards developing salinity tolerant crop plants.

The origin of methylglyoxal in New Zealand manuka ( Leptospermum scoparium) honey

Methylglyoxal in New Zealand manuka honey has been shown to originate from dihydroxyacetone, which is present in the nectar of manuka flowers in varying amounts. Manuka honey, which was freshly produced by bees, contained low levels of methylglyoxal and high levels of dihydroxyacetone. Storage of these honeys at 37 °C led to a decrease in the dihydroxyacetone content and a related increase in methylglyoxal. Addition of dihydroxyacetone to clover honey followed by incubation resulted in methylglyoxal levels similar to those found in manuka honey. Nectar washed from manuka flowers contained high levels of dihydroxyacetone and no detectable methylglyoxal.

Dietary Advanced Glycation Endproducts and Oxidative Stress

Advanced glycation endproducts (AGEs) and oxidative stress (OS) contribute to the development and progression of diabetic complications. We have reported that dietary AGEs and OS induce acute endothelial dysfunction in vivo, but little is known about their effects on adipokines. Twenty inpatients with type 2 diabetes mellitus (mean age: 55.9; range: 32-71 years), received a standard diabetes diet for 6 days. On days 4 and 6, the acute effects of a high-AGE (HAGE) or a low-AGE (LAGE) meal (15.100 vs. 2.750 kU AGE) were studied in a randomized, cross-over, investigator-blinded design. Measurements were performed after an overnight fast, at baseline (B) and at 2, 4, and 6 h after the HAGE or LAGE meals. Both meals had the same ingredients and differed only by the cooking method. Two h following HAGE, a significant decrease from baseline occurred in adiponectin (-10%*double dagger vs. +0%) and leptin (-22%*double dagger vs. -13%*), and a significant increase occurred in vascular cell adhesion molecule 1 (+19%*double dagger vs. -5%) and thiobarbituric acid reactive substances (+23%*double dagger vs. +6%). These changes did not occur, or occurred to a lesser extent, following LAGE. At 4 h following HAGE, an increase in methylglyoxal (+20%double dagger vs. -5%) and E-selectin (+54%*double dagger vs. -3%) occurred. Urinary AGEs increased only after HAGE (+51%*double dagger vs. -2%; values presented as HAGE vs. LAGE; *P < 0.05 vs. baseline, double daggerP < 0.05 vs. LAGE). The postprandial excursions in glucose, insulin, and triglycerides were similar between both meals. A meal rich in AGEs induces acute endothelial and adipocyte dysfunction. These effects were prevented by changing the cooking method.

Methylglyoxal increase in uremia with special reference to snakebite-mediated acute renal failure

BackgroundAdvanced glycation and lipoxidation endproducts (AGEs and ALEs) due to oxidative and carbonyl stress are involved in pathogenesis of several diseases including uremia. Methylglyoxal, a dicarbonyl compound is a metabolic hazard and potent glycating agent in the body, which is an important precursor of AGE and ALE. Methylglyoxal has been reported to be increased in uremia, but there is no report of MG status in snake venom mediated acute renal failure cases (SARF). We investigated the carbonyl and oxidative stress as well as the methylglyoxal concentration in SARF where renal clearance is rapidly shut down. MethodsWe studied serum carbonyl compounds, methylglyoxal, total antioxidant status, GSH and cellular damage marker thiobarbituric acid reacting substances (TBARS) and intracellular erythrocytic GSH concentration following standard methods of 45 SARF and 56 normoglycemic chronic renal failure cases (CRF) and compared with 81 normal controls. ResultMethylglyoxal concentration has been found to be significantly increased in SARF associated with decreased concentration of serum as well as erythrocytic GSH and other antioxidant markers , in comparison with CRF and normal control. The cellular damage (TBARS concentration), is also found increased in SARF. ConclusionMG increase as well as accumulation due to GSH depletion may play a pivotal role in their rapid pathophysiological complicacies in SARF.

Effects of additional nonmethane volatile organic compounds, organic nitrates, and direct emissions of oxygenated organic species on global tropospheric chemistry

This work evaluates the sensitivity of tropospheric ozone and its precursors to the representation of nonmethane volatile organic compounds (NMVOCs) and organic nitrates. A global 3‐D tropospheric chemistry/transport model (IMPACT) has been exercised initially using the GEOS‐Chem chemical reaction mechanism. The model was then extended by adding emissions and photochemical reactions for aromatic and terpenoid hydrocarbons, and by adding explicit representation of hydroxy alkyl nitrates produced from isoprene. Emissions of methanol, phenol, acetic acid and formic acid associated with biomass burning were also added. Results show that O3 increases by 20% in most of the troposphere, peroxyacetyl nitrate (PAN) increases by 30% over much of the troposphere and OH increases by 10%. NOx (NO + NO2) decreases near source regions and increases in remote locations, reflecting increased transport of NOx away from source regions by organic nitrates. The increase in O3 was driven largely by the increased role of PAN as a transporter of NOx and by the rerelease of NOx from isoprene nitrates. The increased PAN production was associated with increases in methyl glyoxal and hydroxyacetone. Comparison with measured values show reasonable agreement for O3 and PAN, but model measurement agreement does not either improve or degrade in the extended model. The extended model shows improved agreement with measurements for methanol, acetic acid and peroxypropional nitrate (PPN). Results from the extended model were consistent with measured alkyl nitrates and glycolaldehyde, but hydroxyacetone and methyl glyoxal were overestimated. The latter suggests that the effect of the isoprene nitrates is somewhat smaller than estimated here. Although the model measurement comparison does not show specific improvements with the extended model, it provides a more complete description of tropospheric chemistry that we believe is important to include.

Gender-related differences in advanced glycation endproducts, oxidative stress markers and nitric oxide synthases in rats

An age- and blood pressure-associated increase in methylglyoxal (MG) and MG-induced advanced glycation endproducts (AGEs), including N(epsilon)-carboxyethyl-lysine (CEL) and N(epsilon)-carboxymethyl-lysine (CML), in the kidney of spontaneously hypertensive rats (SHR) has been shown. In the present study, gender-related changes in AGEs and nitric oxide synthase were investigated in Sprague-Dawley (SD) and stroke-prone SHR (SHRsp) rats. Immunohistochemical analyses were conducted on kidneys from 24-week-old male and female SD rats as well as SHRsp. The systolic blood pressure of SHRsp was significantly higher than that of SD rats. Male SD rats had more intense kidney staining for CEL than female SD rats. Both male and female SHRsp had more marked CEL and CML staining localized to kidney tubules, as opposed to SD rats. Female rats showed more staining in glomerular vessels than male rats in both SD and SHRsp. Nuclei containing nuclear factor-kappaB (NF-kappaB) p65 and activated macrophages were seen in the kidney from SHRsp, not so much in SD rats, localized to renal tubules in male and glomerular vessels in female SHRsp. A higher protein level of NF-kappaB p65 was found in SHRsp than in SD rats. SD rats had more intense kidney neuronal nitric oxide synthase staining than SHRsp. The intensity of inducible nitric oxide synthase staining was significantly higher in SHRsp than in SD rats, with no gender differences in either strain. SHRsp and male rats exhibited higher AGEs and oxidative stress than SD and female rats, respectively. These differences might partly account for the development of hypertension in SHRsp and the higher vulnerability of male animals to renal pathology.

Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress

The mechanism behind enhanced salt tolerance conferred by the overexpression of glyoxalase pathway enzymes was studied in transgenic vis-à-vis wild-type (WT) plants. We have recently documented that salinity stress induces higher level accumulation of methylglyoxal (MG), a potent cytotoxin and primary substrate for glyoxalase pathway, in various plant species [Yadav, S.K., Singla-Pareek, S.L., Ray, M., Reddy, M.K. and Sopory, S.K. (2005) MG levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun. 337, 61–67]. The transgenic tobacco plants overexpressing glyoxalase pathway enzymes, resist an increase in the level of MG that increased to over 70% in WT plants under salinity stress. These plants showed enhanced basal activity of various glutathione related antioxidative enzymes that increased further upon salinity stress. These plants suffered minimal salinity stress induced oxidative damage measured in terms of the lipid peroxidation. The reduced glutathione (GSH) content was high in these transgenic plants and also maintained a higher reduced to oxidized glutathione (GSH:GSSG) ratio under salinity. Manipulation of glutathione ratio by exogenous application of GSSG retarded the growth of non-transgenic plants whereas transgenic plants sustained their growth. These results suggest that resisting an increase in MG together with maintaining higher reduced glutathione levels can be efficiently achieved by the overexpression of glyoxalase pathway enzymes towards developing salinity stress tolerant plants.

A novel HPLC procedure for detection and quantification of aminoacetone, a precursor of methylglyoxal, in biological samples

Increase in methylglyoxal is thought to be involved in different pathological conditions. Deamination of aminoacetone by semicarbazide-sensitive amine oxidase (SSAO) leads to production of methylglyoxal. We have synthesized aminoacetone and developed a novel HPLC procedure for its quantitative determination. The urinary excretion of aminoacetone is approximately 20-30 microg/mouse/day, and the concentration is about 0.5 microg/g in mouse liver and small intestine. SSAO inhibitor increases aminoacetone levels in both tissues and urines. Results confirm that aminoacetone is an endogenous substrate for SSAO. However, data also indicate that deamination is not the only catabolic pathway for aminoacetone.

Ketosis Leads to Increased Methylglyoxal Production on the Atkins Diet

In the popular and widely used Atkins diet, the body burns fat as its main fuel. This process produces ketosis and hence increased levels of beta-hydroxybutyrate (BOB) acetoacetate (AcAc) and its by-products acetone and acetol. These products are potential precursors of the glycotoxin methylglyoxal. Since methylglyoxal and its byproducts are recognized as a significant cause of blood vessel and tissue damage, we measured methylglyoxal, acetone, and acetol in subjects on the Atkins diet. We found that by 14-28 days, methylghyoxal levels rose 1.67-fold (P = 0.039) and acetol and acetone levels increased 2.7- and 6.12-fold, respectively (P = 0.012 and 0.028). Samples from subjects with ketosis showed even greater increases in methylglyoxal (2.12-fold), as well as acetol and acetone, which increased 4.19- and 7.9-fold, respectively; while no changes were seen in samples from noncompliant, nonketotic subjects. The increase in methylglyoxal implies that potential tissue and vascular damage can occur on the Atkins diet and should be considered when choosing a weight-loss program.

Alpha-oxoaldehyde metabolism and diabetic complications.

The factors responsible for variable susceptibility to diabetic nephropathy are not clear. According to the non-enzymatic glycation hypothesis, diabetes-related tissue damage occurs due to a complex mixture of toxic products, including alpha-oxoaldehydes, which are inherently toxic as well as serving as precursors for advanced glycation end-products. Protective mechanisms exist to control this unavoidable glycation, and these are determined by genetic or environmental factors that can regulate the concentrations of the reactive sugars or end-products. In diabetes these protective mechanisms become more important, since glycation stress increases, and less efficient defence systems against this stress could lead to diabetic complications. Some of these enzymatic control mechanisms, including those that regulate alpha-oxoaldehydes, have been identified. We have observed significant increases in production of the alpha-oxoaldehydes methylglyoxal and 3-deoxyglucosone in three human populations with biopsy-proven progression of nephropathy. The increase in methylglyoxal could be secondary to defects in downstream glycolytic enzymes (such as glyceraldehyde-3-phosphate dehydrogenase) that regulate its production, or in detoxification mechanisms such as glyoxalase. Other mechanisms, however, appear to be responsible for the observed increase in 3-deoxyglucosone levels. We present results of our studies on the mechanisms responsible for variable production of alpha-oxoaldehydes by measuring the activity and characteristics of these enzymes in cells from complication-prone and -resistant diabetic patients. New therapeutic interventions designed to control these endogenous mechanisms could potentially enhance protection against excessive glycation and prevent or reverse complications of long-term diabetes.

The Increase of Methyl Glyoxal=like Substance and the Change of Arakawa's Reaction in Rabbits' Milk on an Administration of Monoiod=acetic Acid; Recovery through Vitamin B&lt;sub&gt;1&lt;/sub&gt; and Yakriton

As stated above, all the 4 rabbits used showed methyl glyoxal like substance in the milk, which showed a certain decrease (up to 2.0 mgrms. %) on a subcutaneous use of vitamin B, until the qualitative tests became negative. When monoiodacetic acid was subcutaneously injected, the methyl glyoxal like substance increased gradually, which was proved both quantitatively and qualitatively. Arakawa's reac-tion became weaker and weaker, as the substance increased. Body weight showed a decrease. These changes due to monoiodacetic acid were recovered by vitamin B1 alone when they were of a light degree (as in Rabbit 2), but, when they were of a serious degree (as in Rabbits 3 and 4), only a combined use of yakriton and vitamin B1 was able to improve the condition. It is to be remarked here, especially because Geiger and Rosenberg10) reported that an administration of vitamin B caused a temporary increase of methyl glyoxal in urine of B-avitaminotic dogs, that temporary increase of methyl glyoxal like substance soon after the use of vitamin B1 was seen in the rabbit's milk (Cf. Rabbits 2 and 3) in my own experiment.