There is evidence to suggest that abnormal angiogenesis, inflammation, and fibrosis drive diabetic nephropathy (DN). However, there is no specific treatment to counteract these processes. We aimed to ...determine whether DIAVIT, a natural Vaccinium myrtillus (blueberry) and Hippophae Rhamnoides (sea buckthorn) extract, is protective in a model of type II DN. Diabetic db/db mice were administered DIAVIT in their drinking water for 14 weeks. We assessed the functional, structural, and ultra-structural phenotype of three experimental groups (lean+vehicle, db/db+vehicle, db/db+DIAVIT). We also investigated the angiogenic and fibrotic pathways involved in the mechanism of action of DIAVIT. Diabetic db/db mice developed hyperglycaemia, albuminuria, and an increased glomerular water permeability; the latter two were prevented by DIAVIT. db/db mice developed fibrotic glomeruli, endothelial insult, and glomerular ultra-structural changes, which were not present in DIAVIT-treated mice. Vascular endothelial growth factor A (VEGF-A) splicing was altered in the db/db kidney cortex, increasing the pro-angiogenic VEGF-A165 relative to the anti-angiogenic VEGF-A165b. This was partially prevented with DIAVIT treatment. Delphinidin, an anthocyanin abundant in DIAVIT, increased the VEGF-A165b expression relative to total VEGF-A165 in cultured podocytes through phosphorylation of the splice factor SRSF6. DIAVIT, in particular delphinidin, alters VEGF-A splicing in type II DN, rescuing the DN phenotype. This study highlights the therapeutic potential of natural drugs in DN through the manipulation of gene splicing and expression.
Autism is a behaviorally defined disorder of unknown etiology that is thought to be influenced by genetic and environmental factors. High levels of homocysteine and oxidative stress are generally ...associated with neuropsychiatric disorders. The purpose of this study was to compare the level of homocysteine and other biomarkers in children with autism to corresponding values in age-matched healthy children. We measured total homocysteine (tHcy), vitamin B
12, paraoxonase and arylesterase activities of human paraoxonase 1 (PON1) in plasma and glutathione peroxidase (GPx) activity in erythrocytes from 21 children: 12 with autism (age: 8.29
±
2.76 years) and 9 controls (age: 8.33
±
1.82 years). We found statistically significant differences in tHcy levels and in arylesterase activity of PON1 in children with autism compared to the control group: 9.83
±
2.75 vs. 7.51
±
0.93 μmol/L (
P
≤
0.01) and 72.57
±
11.73 vs. 81.83
±
7.39 kU/L (
P
≤
0.005). In the autistic group there was a strong negative correlation between tHcy and GPx activity and the vitamin B
12 level was low or suboptimal. In conclusion, our study shows that in children with autism there are higher levels of tHcy, which is negatively correlated with GPx activity, low PON1 arylesterase activity and suboptimal levels of vitamin B
12.
Autism spectrum disorders (ASDs), which include the prototypic autistic disorder (AD), Asperger’s syndrome (AS) and pervasive developmental disorders not otherwise specified (PDD‐NOS), are complex ...neurodevelopmental conditions of unknown aetiology. The current study investigated the metabolites in the methionine cycle, the transsulphuration pathway, folate, vitamin B12 and the C677T polymorphism of the MTHFR gene in three groups of children diagnosed with AD (n= 15), AS (n= 5) and PDD‐NOS (n= 19) and their age‐ and sex‐matched controls (n= 25). No metabolic disturbances were seen in the AS patients, while in the AD and PDD‐NOS groups, lower plasma levels of methionine (P= 0.01 and P= 0.03, respectively) and α‐aminobutyrate were observed (P= 0.01 and P= 0.001, respectively). Only in the AD group, plasma cysteine (P= 0.02) and total blood glutathione (P= 0.02) were found to be reduced. Although there was a trend towards lower levels of serine, glycine, N, N‐dimethylglycine in AD patients, the plasma levels of these metabolites as well as the levels of homocysteine and cystathionine were not statistically different in any of the ASDs groups. The serum levels of vitamin B12 and folate were in the normal range. The results of the MTHFR gene analysis showed a normal distribution of the C677T polymorphism in children with ASDs, but the frequency of the 677T allele was slightly more prevalent in AD patients. Our study indicates a possible role for the alterations in one carbon metabolism in the pathophysiology of ASDs and provides, for the first time, preliminary evidence for metabolic and genetic differences between clinical subtypes of ASDs.
Insulin resistance (IR) and cardiometabolic disorders are the main consequences of today's alimentary behavior. This study evaluates the effects of a chronic-discontinuous treatment with alpha-lipoic ...acid (AL), an antioxidant substance that improves glycemic control associated with diabetes mellitus, on metabolic disorders and plasma oxidative stress induced by fructose intake, in rats. Sprague-Dawley rats (48 animals) were randomized into two series (
= 24): rats fed with standard chow or with standard chow supplemented with 60% fructose. In each of the two series, for 2 weeks/month over 12 weeks, a group of rats (
= 12) was intraperitoneally injected with NaCl 0.9%, and a second group (
= 12) received AL 50mg/kg/day. Body weight, glycemia, and systolic blood pressure were monitored throughout the study. After 12 weeks, IR, plasma lipoproteins, uric acid, transaminase activities, and oxidative stress markers were assessed. The high fructose-enriched diet induced cardiometabolic disorders (hypertension, hyperglycemia, IR and dyslipidemia), an increase in uric acid concentration, transaminase activities and C-reactive protein level. This diet also enhanced plasma products of lipid and protein oxidation, homocysteine level, and decreased GSH/GSSG ratio. In this field, there is evidence to indicate that oxidative stress plays an important role in the etiology of diabetic complications. AL discontinuous treatment prevents the metabolic disorders induced by fructose intake, reduced plasma lipid and protein oxidation-products, and restored the GHS/GSSG ratio. Our study proves a promising potential of the chronic-discontinuous treatment of AL and highlights the pleiotropic effects of this antioxidant substance in metabolic disorders such as diabetes.
Introduction: Hyperhomocysteinemia plays an etiologic role in homocystinuria, neurodegenerative and cardiovascular diseases. Potential mechanisms involved in the degenerative diseases of aging ...include: oxidative stress, endothelial dysfunction and inflammation. Aim: In the present study we evaluated the effect of acute administration of homocysteine (Hcy), at a level similar to that found in homocystinuria, on biochemical markers of inflammation (such as IL-6) and of oxidative stress (such as gluthathione peroxidase - GPx). Material and Method: The study was performed on 40 young and older Wistar rats. IL-6 serum level was quantified by a high-sensitivity enzyme-linked immunoabsorbent assay (ELISA) method and the activity of whole blood GPx was measured using a commercially available Randox kit. Results: Our results showed that Hcy administration increased the pro-inflammatory cytokine IL-6 in young rats (p<0.3) and decreased IL-6 level in older rats (p<0.008), when compared to the control group. GPx activity was found to increase with age (587.07 U/gHb versus 847.5 U/gHb, p<0.001). Two hours after Hcy administration, GPx activity was found to decrease, but not in a statistically significant manner. The difference between GPx activities in Hcy treated groups remains statistically significant (p<0.01) in the younger group, compared to older group (556.62 U/gHb versus 748.38 U/gHb). Conclusion: Our results indicate the existence of a correlation between hyperhomocysteinemia, proinflammatory state and oxidative stress, illustrated by the direct dependence of whole blood GPx activities on the increasing age. PUBLICATION ABSTRACT
Autism spectrum disorders (ASD) comprise a complex and heterogeneous group of conditions of unknown aetiology, characterized by significant disturbances in social, communicative and behavioural ...functioning. Recent studies suggested a possible implication of the high‐density lipoprotein associated esterase/lactonase paraoxonase 1 (PON1) in ASD. In the present study, we aimed at investigating the PON1 status in a group of 50 children with ASD as compared to healthy age and sex matched control participants. We evaluated PON1 bioavailability (i.e. arylesterase activity) and catalytic activity (i.e. paraoxonase activity) in plasma using spectrophotometric methods and the two common polymorphisms in the PON1 coding region (Q192R, L55M) by employing Light Cycler real‐time PCR. We found that both PON1 arylesterase and PON1 paraoxonase activities were decreased in autistic patients (respectively, P < 0.001, P < 0.05), but no association with less active variants of the PON1 gene was found. The PON1 phenotype, inferred from the two‐dimensional enzyme analysis, had a similar distribution in the ASD group and the control group. In conclusion, both the bioavailability and the catalytic activity of PON1 are impaired in ASD, despite no association with the Q192R and L55M polymorphisms in the PON1 gene and a normal distribution of the PON1 phenotype.
Paraoxonase 1 (PON1) is an esterase synthesized by the liver and secreted into the plasma, where it is associated with high density lipoproteins (HDL). Its role is to protect LDL and HDL from ...oxidation, thus preventing atherosclerosis. A decreased level of plasma PON1 activities has been found in diabetes mellitus, cardiovascular disorders and chronic liver diseases; but, it can also be influenced by diet and life-style. The purpose of this study was to assess the PON1 activities in the insulin-resistant rats fed with a fructose-enriched diet, in the presence and in the absence of an antioxidant treatment with alpha-lipoic acid (AL).
48 male Sprague-Dawley rats were randomized into two series: rats fed for 3 months with standard chow (Control) or with standard chow supplemented with fructose (60%). In each series, a group of rats was treated intraperitoneally during 14 days/month with NaCl 0.9% and another group with 50mg/kg/day AL. At the end of the 3 months, we assessed: 1) peripheral tissue resistance to insulin (HOMA-IR) and plasma lipid profile, 2) paraoxonase, arylesterase and lactonase activities of PON1, 3) plasma homocysteine (Hcy) level and 4) hepatic transaminase activities: aspartate-aminotransferase and alanine-aminotransferase.
The fructose intake increased peripheral tissue resistance to insulin (HOMA-IR) and plasma lipoprotein level, less the HDL. Also, transaminase and PON1 activities, especially arylesterase and lactonase activities, and the plasma Hcy level were significantly (p>0.05) enhanced in the fructose group. The AL discontinuous treatment associated with the fructose-enriched diet improved the tissue sensitivity to insulin and decreased the plasma lipoprotein levels. Moreover, the AL treatment restored PON1 and transaminase activities, without influencing the Hcy concentration. A decrease in plasma transaminase activities was noted even when AL was associated with standard diet.
In our experimental conditions, the fructose intake induced an increase in plasma transaminase and PON1 activities in association with a Hyperhomocysteinemia. The AL treatment restored the enzymes’ activities and had a hepatoprotective effect, but without influence on Hcy level.
Objectives. The aim of this study was to examine the effects of single nucleotide polymorphisms (SNPs) of PON1 gene at the level of promoter region (‒909 and ‒832) and of first exon (+575, A20352G, ...resulting Q192R substitution) on paraoxonase-1 (PON1) activities in 53 patients with angiographycally proven coronary heart disease (CHD) and 17 free-CHD subjects. Methods and Results. Serum PON1 arylesterase (Ar-ase) and salt-stimulated paraoxonase (ssPO-ase) activities were assessed with manual spectrophotometric methods, by using phenyl acetate and paraoxon as substrates. Common serum biochemical markers were assayed by enzymatic methods using commercial kits, on a Roche/Hitachi 912 Auto Analyzer. PON1 genotypes were determined by PCR and nucleotide sequencing of the amplicons with an ABI PRISMTM 310 Genetic Analyzer and a BigDye® Terminator v3.1 Cycle Sequencing Kit. The severity of coronary artery stenosis was assessed and classified using the Gensini score. We found no significant differences in the PON1 activities and -909(G→C), -832(G→A) and +575(A→G) PON1 polymorphisms between CHD and CHD-free groups. Considering all investigated subjects, we found that -909(G→C) and +575(A→G) SNPs had statistically significant effects on Ar-ase activity and PO-ase activity, respectively. In a multiple regression model we found that diabetes, LDL-cholesterol and the number of mutant alleles were significant independent determinants of the Gensini score. A significant positive correlation was observed only between the Gensini score and the number of mutant alleles. Conclusions. There are no differences between CHD and CHD-free groups regarding PON1 genotypes and phenotypes but the increasing number of PON1 mutant alleles is an important factor in determining the severity of coronary damage.
Obiective. Scopul acestui studiu a fost de a examina efectele polimorfismelor (SNPs) genei PON1 la nivelul regiunii promotor (-909 şi -832) şi a primului exon (+575, 20352A>G, rezultând substituirea Q192R) asupra activităţilor paraoxonazei-1 (PON1) într-un grup format din 53 de pacienţi cu boală coronariană (CHD) dovedită angiografic şi 17 subiecţi fără CHD. Metode şi rezultate. Activităţile PON1 din ser, respectiv arilesteraza (AR-ase) şi paraoxonaza NaCl-stimulată (ssPO-ase), au fost evaluate cu metode spectrofotometrice manuale, folosind acetat de fenil, respectiv paraoxon ca substrat. Markerii biochimici uzuali au fost analizaţi prin metode enzimatice, pe analizorul Roche/Hitachi 912, folosind kituri comerciale. Genotipurile PON1 au fost determinate folosind PCR, iar pentru secvenţierea ampliconilor s-a utilizat kitul BigDye® Terminator v3.1 şi analizorul ABI PRISMTM 310. Severitatea stenozei coronariane s-a evaluat şi clasificat folosind scorul Gensini. Nu s-au observat diferenţe semnificative statistic între grupurile cu şi fără CHD în ceea ce priveste activităţile PON1 şi polimorfismele -909G>C, -832G>A şi +575A>G. Dar, luând în considerare toţi subiecţii investigaţi, am constatat că SNPs -909G>C şi +575A>G au avut efecte semnificative statistic asupra AR-ase, respectiv PO-ase. Folosind modelul regresiei multiple am constatat că diabetul, LDL-colesterolul şi numărul de alele mutante au fost determinanţi semnificativi, independenţi, ai scorului Gensini. O corelaţie pozitivă semnificativă a fost observată numai între scorul Gensini şi numărul de alele mutante. Concluzii. Nu există diferenţe între subiecţi cu si fără CHD în ceea ce priveşte genotipurile şi fenotipurile PON1, dar creşterea numărului de alele PON1 mutante este un factor important în determinarea severităţii leziunilor coronariene
Obiective. Scopul acestui studiu a fost de a examina efectele polimorfismelor (SNPs) genei PON1 la nivelul regiunii promotor (-909 şi -832) şi a primului exon (+575, 20352A>G, rezultând substituirea ...Q192R) asupra activităţilor paraoxonazei-1 (PON1) într-un grup format din 53 de pacienţi cu boală coronariană (CHD) dovedită angiografic şi 17 subiecţi fără CHD. Metode şi rezultate. Activităţile PON1 din ser, respectiv arilesteraza (AR-ase) şi paraoxonaza NaCl-stimulată (ssPO-ase), au fost evaluate cu metode spectrofotometrice manuale, folosind acetat de fenil, respectiv paraoxon ca substrat. Markerii biochimici uzuali au fost analizaţi prin metode enzimatice, pe analizorul Roche/Hitachi 912, folosind kituri comerciale. Genotipurile PON1 au fost determinate folosind PCR, iar pentru secvenţierea ampliconilor s-a utilizat kitul BigDye® Terminator v3.1 şi analizorul ABI PRISMTM 310. Severitatea stenozei coronariane s-a evaluat şi clasificat folosind scorul Gensini. Nu s-au observat diferenţe semnificative statistic între grupurile cu şi fără CHD în ceea ce priveste activităţile PON1 şi polimorfismele -909G>C, -832G>A şi +575A>G. Dar, luând în considerare toţi subiecţii investigaţi, am constatat că SNPs -909G>C şi +575A>G au avut efecte semnificative statistic asupra AR-ase, respectiv PO-ase. Folosind modelul regresiei multiple am constatat că diabetul, LDL-colesterolul şi numărul de alele mutante au fost determinanţi semnificativi, independenţi, ai scorului Gensini. O corelaţie pozitivă semnificativă a fost observată numai între scorul Gensini şi numărul de alele mutante. Concluzii. Nu există diferenţe între subiecţi cu si fără CHD în ceea ce priveşte genotipurile şi fenotipurile PON1, dar creşterea numărului de alele PON1 mutante este un factor important în determinarea severităţii leziunilor coronariene
Hyperhomocysteinemia, an established cardiovascular risk factor, has been recently associated with deep venous thrombosis.
A matched case-control study was designed to assess homocysteinemia as well ...as the acquired risk factors in deep venous thrombosis (DVT). We enrolled 227 subjects, 127 with DVT confirmed by Doppler ultrasonography and 100 controls. Homocysteinemia was measured using reverse-phase high pressure liquid chromatography.
We found a significant association between hyperhomocysteinemia and DVT; the associated risk was weak (p = 0.025, OR: 1.7). Other risk factors significantly associated with DVT were: obesity (p = 0.04, OR for DVT: 2.9), varicose veins (p = 0.023, OR: 3.13), prolonged immobility (p = 0.015, OR: 3.1), history of DVT (p = 0.01, OR: 5.59). All these factors were found to be independent risk factors using multivariate logistic regression.
hyperhomocysteinemia is an independent risk factor for DVT; the risk is not associated with the severity of hyperhomocysteinemia.
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