Chol Group Research Articles

The Effect of Supplementation of Rumen-Protected Choline on Reproductive and Productive Performances of Dairy Cows.

Simple SummaryCholine is a key methyl donor synthesized endogenously in mammals, and its availability is important for various biological functions. A review of the literature presents controversial results and deals with the question of whether rumen-protected choline supplemented during the transition period significantly affects cows’ health, production, and reproduction. However, most recent research shows that in dairy cattle, choline supplementation improves milk yield, composition, and fertility.We aimed to evaluate the effects of organic herbal preparations containing rumen-protected choline (RPC) in dairy cow milk’s BHB and progesterone (P4) concentration changes, reproduction, and production performances. Cows were divided into the following two groups: The CHOL (n = 60) cow diet was supplemented with 10 g/day RPC from 20 days pre-calving to 20 days post-calving, and CONT (n = 60) were fed a conventional diet. BHB and P4 concentrations were measured at 5–64 DIM and 21–64 DIM, respectively, with DelPro 4.2. BHB was lower in the CHOL group at 5–64 DIM than CONT p > 0.05. The first post-calving P4 peak, p < 0.001, was determined earlier in the CHOL group, and the P4 profile during 21–64 DIM was similar, p > 0.05. The insemination rate was lower, and the interval between calvings was shorter. The first insemination time was earlier in the CHOL group, p < 0.05. Milk yield was higher in the CHOL group at 21–64 DIM, p > 0.05. The CHOL group had more fat in their milk at 31–60 DIM, p < 0.05. There were no significant differences in protein and SCC between the groups, p > 0.05. Based on our results, we concluded that the supplementation of RPC pre- and post-calving had statistically significant effects on the first peak of P4, and benefited the reproduction performances, milk yield, and milk fat during the early postpartum period.

Cholesterol Induces Oxidative Stress, Mitochondrial Damage and Death in Hepatic Stellate Cells to Mitigate Liver Fibrosis in Mice Model of NASH.

Liver fibrosis and its end-stage disease cirrhosis are major world health problems arising from chronic injury of the liver. In recent years, the hypothesis that hepatic stellate cells’ (HSCs’) activation and fibrosis can be mitigated by HSC apoptosis and cell death has become of interest. In the current study, we evaluated the effect of cholesterol and bile acids on HSC apoptosis and liver fibrosis. Male C57BL/6J mice (wild type), aged four to five weeks, were fed an AIN-93G based diet (normal diet, ND), ND diet + 1% (w/w) cholesterol (CHOL group), ND diet + 0.5% (w/w) cholic acid (CA group) or ND diet + 1% (w/w) cholesterol + 0.5% (w/w) cholic acid (CHOL + CA group). Female Mdr2(-/-) mice were also treated with ND with and without 1% cholesterol. The effect of cholesterol on liver fibrosis and HSC clearance was evaluated. In addition, we studied the mechanism of cholesterol-induced apoptosis in HSC-T6 and AML-12 hepatocyte cell lines. In animals treated with cholic acids, increased lipid peroxidation and fibrosis were observed after six weeks of treatment. However, addition of cholesterol to the diet of C57BL/6J mice led to HSC-specific apoptosis and resolution of liver fibrosis, verified by double-staining with active caspase and α smooth muscle actin antibodies. In Mdr2 (-/-) mice, a diet supplemented with cholesterol corrected fibrosis and induced active hepatic stellate cells’ clearance. HSC-T6 were found to be much more sensitive to cholesterol-induced oxidative stress, mitochondrial damage and apoptosis compared to hepatocytes. These results indicate that cholesterol may be a trigger of HSC lipid peroxidation and death in the liver in a model of non-alcoholic steatohepatitis. A high cholesterol-to-bile acid ratio may determine the trajectory of the liver disease toward mitigation of fibrosis.

Cornelian cherry extract ameliorates osteoporosis associated with hypercholesterolemia in New Zealand rabbits.

Results of animal studies show that a high-cholesterol diet increases bone resorption and decreases bone formation, thus leading to osteoporosis. Previously, we reported on the beneficial influence of Cornelian cherry (Cornus mas L.) fruit on lipid profile in an animal model of diet-induced hipercholesterolemia. To investigate the influence of Cornus mas L. extract and loganic acid (LA) on cholesterol-induced bone changes. The study was conducted on 50 New Zealand rabbits. The animals were given either standard chow (group P) or the same standard chow enriched with 1% cholesterol (other groups). Additionally, the group CHOL+EX received Cornus mas L. extract, group CHOL+LA - loganic acid, and group CHOL+SIM - simvastatin. Serum concentration of bone turnover markers, bone mineral density (BMD) and bone micro-computed tomography (microCT) were assessed. In the CHOL group, a decrease in osteocalcin (OC) and an increase in C-terminated telopeptide of type I collagen (CTX) levels were detected (CHOL vs P 0.674 ±0.159 ng/mL vs 1.003 ±0.297 ng/mL and 10.049 ±1.276 ng/mL vs 7.721 ±1.187 ng/mL, respectively). The EX and LA ameliorated cholesterol-induced changes in serum OC (0.857 ±0.160 ng/mL and 1.103 ±0.356 ng/mL, respectively) and CTX (7.735 ±1.045 ng/mL and 8.128 ±1.106 ng/mL, respectively). There was a significant decrease in femoral BMD in CHOL group (0.429 ±0.11 g/cm² vs 0.449 ±0.020 g/cm²). The EX and LA ameliorated those changes (0.458 ±0.016 g/cm² and 0.449 ±0.021 g/cm², respectively). The microCT revealed increased bone volume ratio (BV/TV) and trabecular thickness (Tb.Th.) in the CHOL+EX group. Cornus mas L. inhibited bone resorption and stimulated bone formation, thereby preventing the development of cholesterol-induced osteoporosis.

Cholesterol, a repressor of natriuretic proteins

Salt‐sensitivity in Dahl rats is due, in part, to the reduction of hemoxygenase‐1 (HO1), cyclooxygenase 2 (COX2), and nitric oxide synthase‐2 (NOS2) in the renal medulla. Renal overexpression of HIF1‐α, a regulator of HO1, COX2, and NOS2, enhances salt and water excretion and reduces blood pressure. Cholesterol (chol) incorporation into collecting duct (CD) cells represses COX2 activity in response to tubular flow or fluid shear stress (FSS). We hypothesized that dietary chol ingestion represses flow responsive a genes necessary to effectuate salt and water excretion.To this end, mice were fed either a 0% or 1% chol diet for up to 12 weeks, injected with SQ isotonic saline at 5% of body weight, and urine collected at 2, 4, and 6 hrs. Kidneys were extracted to measure medullary expression of HO1, COX1, and NOS2. Mice were divided into three dietary time points: (1) 3–5 (2) 6–8 and (3) &gt;9 weeks of diet.Urine volume was less in the chol group at the 6 hr collection for both 6–8 (n=8 each group; p&lt;0.05) and &gt;9 week (n=8 each group, p&lt;0.05) groups. Urinary [Na], [K] and osmolality were higher while Na excretion was lower at the 6 hr urine collection time point in the chol (n=8; p&lt;0.05) vs. control (n=8) fed mice at 6–8 week group.. Renal medullary HO1 (0.76±0.09; p&lt;0.05), and NOS2 (0.76±0.1; p&lt;0.05) mRNA expression were down regulated compared to controls while COX2 and HIF1‐α were unaffected. ATP‐binding cassette transporter (ABCA1), a tissue chol efflux transporter, was increased in the medulla (2.0±0.3; p&lt;0.05) of chol fed mice. To test the flow responsiveness of HO1, COX2, NOS2 and HIf1‐α, IMCD3 cells were exposed to 0.4 dynes/cm2 of FSS for up to 6 hours. HO1 mRNA increased by 350X fold, COX2 by 25X fold, and NOS2 by 8X fold at 6 hrs in sheared (n=4–6; p&lt;0.01) vs. static (n=3–6) cells. HIF1‐α mRNA increased by ~50% (p&lt;0.05) at 4 and 6 hrs. Western blotting of FSS exposed cells increased the steady state protein abundance of NOS2 at 2, 4, and 6 hrs vs. static controls, while HO1 and COX2 protein abundance increased at 4 and 6 hrs vs. static controls. In contradistinction, HIF1‐a protein expression was reduced at 2, 4, and 6 hrs in FSS exposed cells vs static controls. Next, shear exposed, chol loaded IMCD3 cells were compared to shear exposed, untreated IMCD3 cells to test whether cholloading represses FSS sensitive natriuretic protein expression. At 2 hrs of FSS, COX2 (−29%), NOS2 (−23%) and HO1 (−18%) protein expression was significantly (p&lt;0.05) but modestly reduced in chol loaded cells vs. untreated controls. Similar to the mouse, chol incorporation into cells raised ABCA1 mRNA levels 3.7±0.2 fold vs. untreated cells.Dietary chol is associated reduced urine volume and Na excretion at 6 hrs after saline injection and suppressed flow induced HO1 and NOS2 gene expression. In vitro studies of cultured IMCD3 cells illustrate that FSS induce COX2, NOS2 and HO1 expression, and that chol loading of cells can suppress this effect. We speculate that chol incorporation into tubular epithelia represses of natriuretic factors necessary excrete Na.Support or Funding InformationVAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effects of the Dietary Addition of Amaranth (Amaranthus mantegazzianus) Protein Isolate on Antioxidant Status, Lipid Profiles and Blood Pressure of Rats.

The effects of the dietary addition of 2.5% (w/w) Amaranthus mantegazzianus protein isolate (AI) on blood pressure, lipid profiles and antioxidative status of Wistar rats were evaluated. Six diets were used to feed animals during 28 days: (base (AIN93G), Chol (cholesterol 1%, w/w), CE (α-tocopherol 0.005%, w/w), CholE (cholesterol 1% (w/w) + α-tocopherol 0.005%, w/w), CAI (AI 2.5% w/w), CholAI (cholesterol 1% (w/w) + AI 2.5%, w/w). Lipid profiles of plasma and liver and faecal cholesterol content were analyzed. Antioxidant status was evaluated by the ferric reducing activity of plasma (FRAP), the 2-thiobarbituric acid (TBA) assay and superoxide dismutase (SOD) activity in plasma and liver. Blood pressure was measured in the tail artery of rats. CholA group presented a significant (α < 0.05) reduction (16%) in the plasma total cholesterol. In liver, the intake of cholesterol (Chol group) induced a significant increment in cholesterol and triglycerides (2.5 and 2.3 times, respectively), which could be decreased (18% and 47%, respectively) by the addition of AI (CholA group). This last group also showed an increased faecal cholesterol excretion (20%). Increment (50%) in FRAP values, diminution of TBA value in plasma and liver (70% and 38%, respectively) and diminution of SOD activity (20%) in plasma of CholA group suggest an antioxidant effect because of the intake of AI. In addition, CA and CholA groups presented a diminution (18%) of blood pressure after 28 days.

Rapana venosa consumption improves the lipid profiles and antioxidant capacities in serum of rats fed an atherogenic diet

In the recent years, the consumption of seafood has increased. There are no results on the studies of Rapana venosa (Rv) as a supplementation to the diets. We hypothesized that Rv would increase antioxidant capacity and reduce blood lipids, based on the composition of bioactive compounds and fatty acids. Therefore, the aim of this investigation was to evaluate in vitro and in vivo actions of Rv from contaminated (C) and non-C (NC) regions of collection on lipid profiles, antioxidant capacity, and enzyme activities in serum of rats fed an atherogenic diet. Twenty-four male Wistar rats were divided into 4 groups of 6 each and named control, cholesterol (Chol), Chol/RvC and Chol/RvNC. Rats of all 4 groups were fed the basal diet, which included wheat starch, casein, soybean oil, cellulose, vitamin (American Institute of Nutrition for laboratory animals vitamin mixtures), and mineral mixtures (American Institute of Nutrition for laboratory animals mineral mixtures). During 28 days of the experiment, the rats of the control group received the basal diet only, and the diets of the other 3 groups were supplemented with 1% of Chol, 1% of Chol, and 5% of Rv dry matter from C and NC areas. Dry matter from C and NC areas supplemented diets slightly hindered the rise in serum lipids vs Chol group: total Chol, 13.18% and 11.63% and low-density lipoprotein Chol, 13.57% and 15.08%, respectively. Cholesterol significantly decreased the value of total antioxidant capacity. The supplementation of Rv to the Chol diet significantly affected the increase of antioxidant capacity in serum of rats, expressed by the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) method. The water extracts of Rv exhibited high binding properties with bovine serum albumin in comparison with quercetin. In conclusion, atherogenic diets supplemented with Rv from C and NC areas hindered both the rise in serum lipids levels and the decrease in the antioxidant capacity. Based on fluorescence and electrospray-ionization mass spectrometry profiles and in vivo studies, changes in the intensity of the found peaks were estimated in the serum samples after supplemented diets. These findings indicate that the supplementation of Rv to the atherogenic diets improve the lipid profiles and the antioxidant status in serum of rats.

The natural olive constituent oleuropein induces nutritional cardioprotection in normal and cholesterol-fed rabbits: comparison with preconditioning.

Ischemic preconditioning, which is mediated by cell signaling molecules, protects the heart from ischemia-reperfusion injury by limiting the infarct size. Oleuropein, the main polyphenolic constituent of olives, reduced the infarct size in normal and cholesterol-fed rabbits when it was administered at a nutritional dose. The aim of the present study was to compare the effects of oleuropein and preconditioning in terms of the cell signaling and metabolism pathways underlying myocardial protection. Rabbits were randomly divided into six groups: the control group received 5 % dextrose for six weeks, the preconditioning group was subjected to two cycles of preconditioning with 5 min ischemia/10 min reperfusion, the O6 group was treated with oleuropein for six weeks, the Chol group was fed a cholesterol-enriched diet and 5 % dextrose for six weeks, and the CholO6 and CholO3 groups were treated with cholesterol and oleuropein for six and three weeks, respectively; oleuropein was dissolved in 5 % dextrose solution and was administered orally at a dose of 20 mg × kg(-1) × day(-1). All animals were subsequently subjected to 30 min myocardial ischemia followed by 10 min of reperfusion. At that time, myocardial biopsies were taken from the ischemic areas for the assessment of oxidative and nitrosative stress biomarkers (malondialdehyde and nitrotyrosine), and determination of phosphorylation of signaling molecules involved in the mechanism of preconditioning (PI3K, Akt, eNOS, AMPK, STAT3). The tissue extracts NMR metabolic profile was recorded and further analyzed by multivariate statistics. Oxidative biomarkers were significantly reduced in the O6, CholO6, and CholO3 groups compared to the control, preconditioning, and Chol groups. Considering the underlying signaling cascade, the phosphorylation of PI3K, Akt, eNOS, AMPK, and STAT-3 was significantly higher in the preconditioning and all oleuropein-treated groups compared to the control and Chol groups. The NMR-based metabonomic study, performed through the analysis of spectroscopic data, depicted differences in the metabolome of the various groups with significant alterations in purine metabolism. In conclusion, the addition of oleuropein to a normal or hypercholesterolemic diet results in a preconditioning-like intracellular effect, eliminating the deleterious consequences of ischemia and hypercholesterolemia, followed by a decrease of oxidative stress biomarkers. This effect is exerted through inducing preconditioning-involved signaling transduction. Nutritional preconditioning may support the low cardiovascular morbidity and mortality associated with the consumption of olive products.

Cholesterol- and lanolin-rich diets may protect against steroid-induced osteonecrosis in rabbits

Background and purpose It remains controversial how hypercholesterolemia influences the development of steroid-induced osteonecrosis (ON). We investigated the role of hypercholesterolemia induced by a cholesterol-rich diet on the development of ON in rabbits.Methods 40 adult male Japanese white rabbits were randomly divided into 2 groups. 20 rabbits were maintained on a cholesterol-rich diet for 2 weeks before receiving steroid treatment (the CHOL group). The other 20 rabbits were maintained on a standard diet (the control (CTR) group). 2 weeks after the start of the study, all 40 rabbits were injected with methylprednisolone acetate (MPSL) into the right gluteus medius muscle (20 mg/kg body weight). 2 weeks after the steroid injection, both the femora and humeri were examined histopathologically for the presence of ON. Hematological analysis of the serum lipid levels was performed every week. Based on the same protocol, we also investigated the effects of lanolin, a primary component of a cholesterol-rich diet, in another group (the LA group).Results The incidence of ON in the CHOL group (3/20) was lower than that observed in the CTR group (15/20) (p < 0.001). During the whole experiment, the levels of total cholesterol and the ratio of low-density lipoprotein to high-density lipoprotein in the CHOL group were higher than those observed in the CTR group (p < 0.001). The LA group also had a lower incidence of ON (2/20), and the lipid levels in the LA group showed similar changes to those observed in the CHOL group.Interpretation Our findings suggest that preexisting hypercholesterolemia itself induced by a cholesterol-rich diet does not increase the risk of developing steroid-induced ON, but rather seems to diminish it. Lanolin may be the active anti-ON component of the cholesterol diet.

Comparative Study of Tomato and Tomato Paste Supplementation on the Level of Serum Lipids and Lipoproteins Levels in Rats Fed With High Cholesterol

BackgroundHypercholesterolemia is one of the risk factors of cardiovascular diseases. Increased blood cholesterol affects general health and increases mortality due to cardiovascular disease. Poor nutrition increases LDL cholesterol and decreases LDL receptor activities in the liver. Scientists have shown that consumption of antioxidants can reduce hypercholesterolemia and proved benefits of fruit and vegetables. Tomato reduces oxidative stress by increasing serum total antioxidant level.ObjectivesThis study compared the tomato and tomato paste supplementation on the level of serum lipids and lipoproteins in rats fed with high cholesterol.Materials and MethodsIn this study, four male rat groups (10 rats per group) were used. Control group received basal diet, second group received basal diet and 2% cholesterol (Chol), third and fourth groups received basal diet, 2% cholesterol tomato and tomato paste respectively (20 percent of the diet) for a month. Then serum TC, LDL, HDL and TG were measured.ResultsResults showed that in Chol group, all lipids increased significantly (P < 0.05) except HDL compared to the control group. Tomato and tomato paste supplementation decreased TC, LDL and TG concentration significantly (P < 0.05) compared to Chol group. Tomato paste had the higher effect on lipids decreasing than tomato.ConclusionsDecreases of TC, LDL and TG may be related to tomato antioxidant effect. This course in human required more investigations.

Juniperus communis Linn oil decreases oxidative stress and increases antioxidant enzymes in the heart of rats administered a diet rich in cholesterol

It has been asserted that consumption of dietary cholesterol (Chol) raises atherosclerotic cardiovascular diseases and that Chol causes an increase in free radical production. Hypercholesterolemic diet has also been reported to cause changes in the antioxidant system. In our study, different doses of Juniperus communis Linn (JCL) oil, a tree species growing in Mediterranean and Isparta regions and having aromatic characteristics, were administered to rats; and the levels of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and thiobarbituric acid reactive substances assay (TBARS) were examined in the heart tissue of rats. In this study, 35 Wistar Albino male adult rats weighing approximately 250-300g were used. The rats were divided into five groups of seven each. The control group was administered normal pellet chow, and the Chol group was administered pellet chow including 2% Chol, while 50 JCL, 100 JCL, and 200 JCL groups were administered 50, 100, and 200mg/kg JCL oil dissolved in 0.5% sodium carboxy methyl cellulose, respectively, in addition to the pellet chow containing 2% Chol, by gavage. After 30days, the experiment was terminated and the antioxidant enzyme activities were examined in the heart tissue of rats. While consumption of dietary Chol decreases the activities of SOD, GSH-Px, and CAT in heart tissue of rats (not significant), administeration of 200mg/kg JCL oil in addition to Chol led to a significant increase in the activity of antioxidant enzymes. Administering Chol led to a significant increase in TBARS level. Administering 100 and 200mg/kg JCL oil together with Chol prevented significantly the increase in lipid peroxides. As a result of the study, JCL oil showed oxidant-antioxidant effect in the heart tissue of rats.

Investigating the effectiveness of different tea types from various thyme kinds (Origanum onites, Thymbra spicata and Satureja cuneifolia) on anemia and anticholesterolemic activity

In a study on villagers settled on the outskirts of the Taurus Mountains and whose source of living is thyme, it was revealed that the villagers excessively consumed thyme by adding it to their tea and many of their foods; high incidences of anemia was found among these villagers. In this study, 42 male adult Wistar albino rats weighing 200-250g were used. The rats were divided to six equal groups as follows: control, cholesterol (Chol), 80mg/kg Origanum onites Labiatae (OOL), 80mg/kg Thymbra spicata Labiatae (TSL), 80mg/kg Satureja cuneifolia Labiatae (SCL), and 160mg/kg TSL, and each group consisted of seven rats. The control group was fed with normal pellet feed. The Chol group and all the other groups, except for the control group, were fed with 2% cholesterol-containing pellet feed. Physiological serum of 4ml was given to the control and Chol group, wheile 80 mg/kg of thymes tea was given to the OOL group, TSL group, and SCL group, and 160mg/kg of thymes tea was given to the TSL group by means of a gavage for 30days. In the blood samples, the hematologic parameters and the biochemical parameters of serum glucose, blood urea nitrogen, creatinine, total protein, albumin, iron (I), total iron-binding capacity, aminotransferase aspartate, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein cholesterol, triglyceride, and oxidized LDL levels were examined. The kidney and liver tissues were examined histopathologically. The results of the study showed that different types of thymes had an antihypercholesterolemic effect. In addition to the anemic effect detected in group TSL and the mild granular degeneration found in the liver of 80mg/kg SCL group, distinct granular degeneration was observed in 160mg/kg TSL group.

A NOVEL VWF A3-GPI MODIFIED EPCS TO ENHANCE ITS ADHESION ABILITY TO DAMAGED VESSEL SEGMENT-COLLEGEN

ObjectivesThis study was designed to investigate the relationship between hame oxygenase (HO)/carbon monoxide (CO) system and nitric oxide synthase (NOS)/nitric monoxide (NO) system in diet-induced atherosclerosis.MethodsThe rabbits serving as animal...

THE TIMELINESS RESEARCH OF OXIDATIVE/ANTI-OXIDATIVE SYSTEM DURING THE RABBIT ATHEROSCLEROTIC MODELLING

ObjectivesThe purpose of the present study was to investigate the effect of different enzymes on the anti-oxidative defense system and oxidative stress in the rabbit model of atherosclerosis.MethodsTwenty four male...

Alteration of Some Inflammatory Biomarkers by Dietary Oxysterols in Rats

Oxysterols are structurally similar to cholesterol, but are characterized by one or more additional oxygen-containing functional groups. These compounds are implicated in inflammation given their ability to cause irreversible damage to vascular cells. The aim of this study was to study the alteration of some inflammatory biomarkers in Wistar rats in response to dietary oxysterols. Eighteen rats were randomly divided into three groups of six rats each. A standard diet supplemented with 1% (w/w) pure cholesterol (Chol group) or 1% (w/w) of an oxidized cholesterol mixture (COPs group) was fed for 8 weeks. Blood serum was separated; abdominal, pericardial, and epididymal adipose tissue was removed carefully. The COPs subjects exhibited significant increase in blood pressure and serum triacylgycerols as well as increased body fat index and pericardic, abdominal, and epididymal adipose tissue. These effects were accompanied by elevated circulating levels of plasma high-sensitivity C-reactive protein, tumor necrosis factor alpha, and resistin. We suggest that dietary oxysterols have an important pro-inflammatory effect.

KB-R7943 Reduces Necrosis and Apoptosis in Hyperlipidemic Animals through the Activation of K+ATP Channels

Reverse-mode activation of the Na+-Ca2+ exchanger (NCX) at the time of reperfusion following ischemia contributes to Ca2+ overload and cardiomyocyte injury, KB-R7943, as a selective reverse-mode NCX inhibitor, reduces lethal reperfusion injury under normal conditions, but its effectiveness under certain pathological states is in dispute. In the present study, we sought to determine the effect of KB-R7943 in hyperlipidemic animals and assess if the K+ ATP are involved in the protective mechanisms. Anesthetized rats were randomized into five groups, as follows, and were subjected to 25min global ischemia (GI) followed by 120 min reperfusion (R). Normally fed animals: a Control group (NC) with no additional intervention. Cholesterol fed (6weeks) animals: a Chol group (HC) with no additional interventions, a KB-R7943 group (HKB) treated with KB-R7943, a KB-R7943 and glybenclamide group (HGKB) treated with KB-R7943 and glybenclamide (K+ ATP, blocker, 0.3μM), a glybenclamide group (HGLY) treated with glybenclamide The infarct size was measured by triphenyltetrazolium. The infarct size was 35±5.0% in NC, 46±8.7% in HC and 47±8.5% in HGLY (NC vs. HC, P 0.05). KB-R7943 reduced the infarct size (28.6±3.3% in HKB vs. HC, P<0.05), In addition, KB-R7943 attenuated apoptotic cell (HKB vs. HC, p<0.05), glybenclamide abolished the effect reached by KB-R7943. Thus, diet-induced hypercholesterolemia enhances myocardial injury; KB-R7943 reduces infarct size and apoptosis in hyperlipidemic animals through the activation of K+ ATP channels.

Dietary red palm oil supplementation decreases infarct size in cholesterol fed rats

Background and AimsThe effect of red palm oil (RPO) supplementation on infarct size after ischaemia/reperfusion in a cholesterol enriched diet-induced hyperlipidemic animal model has not been reported. Previous studies reported results on the effect of RPO in a normal diet, whilst evidence of protection has been linked to improved functional recovery, prosurvival kinase, anti-apoptosis and NO-cGMP. Therefore, we aimed to investigate the effects of dietary RPO supplementation in a cholesterol-enriched diet-induced hyperlipidemic rat model and to investigate the involvement of matrix metalloproteinase 2 (MMP2) inhibition as a possible mechanism of protection.Materials and MethodsMale Wistar rats were fed either a standard rat chow diet (Norm) or a 2% cholesterol-enriched diet (Chol) for nine weeks. Additionally, two more groups received the same treatment, however, at the week 4, diet was supplemented with RPO for the last five weeks (Norm+RPO and Chol+RPO), respectively. After the feeding period hearts were isolated, perfused according to Langendorff and subjected to 30 minutes of normothermic global ischaemia followed by two hours of reperfusion. Infarct size was measured by 2,3,5-triphenyltetrazolium chloride staining at the end of reperfusion.ResultsCholesterol-enriched diet increased myocardial infarct size from 23.5 ± 3.0% to 37.2 ± 3.6% (p < 0.05) when compared to normal diet. RPO supplementation significantly reduced infarct size either in Norm+RPO or in Chol+RPO (to 9.2 ± 1.0% and 26.9 ± 3.0%), respectively. Infarct size in Chol+RPO was comparable to the Norm group. MMP2 activity before ischaemia was significantly reduced in the Chol+RPO group when compared to the Chol group. However, the MMP2 activity of the hearts of the RPO fed rats was significantly increased when compared to the normal diet group after ischaemia.ConclusionsFor the first time it was shown that dietary RPO supplementation attenuated the increased susceptibility of the hearts in cholesterol fed rats to ischaemia/reperfusion injury. This was shown by reduced infarct size. For the first time we also show that red palm oil supplementation altered pre-ischaemic levels of MMP-2, which may indicate that myocardial MMP2 may be implicated as a possible role player in RPO mediated protection against ischaemia/reperfusion injury in hearts of cholesterol supplemented rats.

Aorta and Liver Changes in Rats Fed Cholesterol-Containing and Raw Vegetable-Supplemented Diets: Experiments in Vitro and in Vivo

The aim of this investigation was to compare the liver and aorta changes in rats fed cholesterol-containing diets and the possible improvement when diets would be supplemented with frequently used raw vegetables. The phenolic compounds of three vegetables in methanol-water (1:1) fraction were characterized using electrospray ionization mass spectra (ESI-MS). Results showed that the content of polyphenols, flavonoids, quercetin, flavanols, tannins, and ascorbic acid varied for garlic and white and red onions ranging from 6.68 to 18.08 mg GAE/g DW, 490.4-701.0 μg CE/g DW, 281.2-1100.0 μg, 32.40-41.30 μg CE/g DW, 2.88-3.12 mg CE/g DW, 1.87-2.33 mg AA/g DW, 1388.2-1442.3 μg CGE/g DW, respectively. The radical scavenging capacities (μM TE/g DW) for the same investigated vegetables for ABTS, FRAP, CUPRAC, and DPPH assays ranged from 48.78 to 92.42, 9.41-28.56, 3.06-10.41, and 6.49-23.42, respectively. Good correlations were observed between the phenolic contents and the radical scavenging capacities of the vegetables. The interaction between BSA and quercetin, BSA and garlic and onions extracts was measured by 3-dimensional fluorescence (3D-FL) and Fourier transform infrared (FT-IR) spectroscopy. The highest polyphenol content was found in methanol/water fraction of onions and garlic; therefore, for the investigation of in vitro interactions with BSA only polyphenols of this fraction were used. For in vivo studies, 30 male Wistar rats were divided into 5 groups each of 6 and named Control, Chol, Chol/Garlic, Chol/OnionRed, and Chol/OnionWhite. During 6 weeks, the rats of all 5 groups were fed a basal diet (BD). The rats of the Control group were fed the BD only. The BD of the Chol group was supplemented with 10 g/kg of nonoxidized cholesterol (NOC). Each of the other three groups was supplemented with 10 g/kg of NOC and 500 mg of raw fresh garlic, 500 mg of raw fresh red onion, and 500 mg of raw fresh white onion on 1 kg of body weight for Chol/Garlic, Chol/OnionRed, and Chol/OnionWhite diet groups, respectively. In order to detect the changes in the liver and aorta, a histological procedure was applied, and the liver enzymes were determined and compared. It was found that the main changes vs the Control group were in the liver of rats fed the cholesterol-containing diet without vegetable supplementation. Significantly less histological changes in the liver and lower level of liver enzymes vs those of the Chol group were detected in rats of the Chol/Garlic group (P < 0.05). The interaction between the polyphenol extract of garlic and BSA in vitro showed its strong ability comparable with that of quercetin to quench the intrinsic fluorescence of BSA. In conclusion, all studied vegetables showed protective effects, but raw garlic supplemented with cholesterol-containing diets significantly prevented the aorta and liver damages of rats.

Influence of two cultivars of persimmon on atherosclerosis indices in rats fed cholesterol-containing diets: Investigation in vitro and in vivo

Objective To assess the influence of two persimmon cultivars on some atherosclerosis indices in rats fed cholesterol (Chol)-containing diets. Methods Persimmon cultivars “Fuyu” and “Jiro” as supplementation to rats’ diets were investigated in vitro to compare the contents of their bioactive compounds (polyphenols, flavonoids, flavanols, tannins, carotenoids, and ascorbic acid) and antioxidant potentials. In the in vivo investigation, 36 male Wistar rats were randomly divided into six diet groups, each with six rats: control, control/Fuyu, control/Jiro, Chol, Chol/Fuyu, and Chol/Jiro. During a period of 47 d (42 d of feeding and 5-d adaptation before the experiment) of the trial, rats in the control group were fed a basal diet and two additional control groups (control/Fuyu and control/Jiro) a basal diet plus 5% of lyophilized Fuyu and Jiro, respectively. The Chol, Chol/Fuyu, and Chol/Jiro rat groups were fed a basal diet supplemented with 1% Chol (Chol group) and 1% Chol plus 5% lyophilized Fuyu (Chol/Fuyu group) and plus 5% lyophilized Jiro (Chol/Jiro group), respectively. After completion of the experiment, the rats were anesthetized using Narcotan (halothane) and sacrificed and the atherosclerotic lesions in the aorta were assessed. The obtained results of the investigation of all six groups were compared. Testing of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triacylglycerols, total cholesterol in the liver, electrophoretic patterns of liver tissue, and three-dimensional fluorescence of serum protein fractions was performed. Results The polyphenols and tannins were significantly higher in the Fuyu cultivar ( P < 0.05). The antioxidant potential of persimmon Fuyu was higher than in the Jiro cultivar, but the difference was significant only according to the 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay ( P < 0.05). Supplementation of diets with 5% of the lyophilized Fuyu and Jiro hindered the increase in plasma lipids versus the Chol group (total cholesterol 19.4% and 9.5%, low-density lipoprotein cholesterol 25.6% and 13.1%, respectively, P < 0.05) and hindered the decrease in plasma antioxidant activity versus the Chol group by 40.0% and 16.8% and by 39.6% and 11.3% for the ABTS and 1,1-diphenyl-2-picrylhydrazyl assays, respectively. The atherosclerotic lesions in the aortas of the Chol/Fuyu and Chol/Jiro groups were significantly less than in the Chol group ( P < 0.05). Electrophoresis of the proteins from rats’ liver tissue showed changes in 14-kDa bands after persimmon supplementation. A shift in maximum wavelengths in three-dimensional fluorescence of serum protein fractions after persimmon supplementation was found in comparison with the control group and an increase in fluorescence intensity compared with the Chol groups. Conclusion The contents of polyphenols and tannins were significantly higher in the Fuyu cultivar ( P < 0.05). The antioxidant potentials of Fuyu were higher than those of the Jiro cultivar, but the difference was significant only according to the ABTS assay ( P < 0.05). Supplementation of 5% lyophilized Fuyu and Jiro to diets of rats fed Chol-containing diets 1) hindered the increase in plasma lipids levels and the decrease in plasma antioxidant activity and 2) significantly decreased the atherosclerotic lesions in the aorta ( P < 0.05). Electrophoretic patterns of liver tissue and fluorescence spectra can be used as additional biomarkers for determination of atherosclerosis indices.

Antioxidant properties and bioactive constituents of some rare exotic Thai fruits and comparison with conventional fruits: In vitro and in vivo studies

The aim of this research was to investigate the bioactivity of durian, snake fruit and mangosteen, rare exotic Thai fruits. These fruits were compared among them and with conventional fruits: durian with mango and avocado, and snake fruit with mangosteen and kiwifruit in order to find the preferable diet for human consumption. The contents of polyphenols, flavonoids, flavanols, tannins, anthocyanins, ascorbic acid and carotenoids, and the level of antioxidant potential by ABTS, DPPH, FRAP and CUPRAC in different extracts (methanol, water, acetone, and hexane) were determined. The presence of polyphenols (flavonoids and phenolic acids) in the investigated samples was characterized by Fourier transform infrared (FT-IR) spectroscopy and three-dimensional fluorimetry (3D-FL).The in vivo studies were carried out on 25 male Wistar rats, divided into 5 diet groups, each of 5. During 30days of the experiment the rats of all 5 groups were fed basal diet (BD), which included wheat starch, casein, soybean oil, vitamin and mineral mixtures. The rats of the Control group were fed only the BD. The BD of the other 4 groups was supplemented with 1% of nonoxidized cholesterol (NOC) (Chol group), 1% of NOC in each group and 5% of lyophilized fruits: durian (Chol/Durian), snake fruit (Chol/Snake), mangosteen (Chol/Mangosteen). After the experiment diets supplemented with exotic fruits significantly hindered the rise in plasma lipids and hindered the decrease in the plasma antioxidant activity. In conclusion, the contents of bioactive compounds and the antioxidant potential are relatively high in the studied fruits and varied among them depending on the extraction procedure. FT-IR and 3D-FL can be used as additional tools for identification and comparison of bioactive compounds. Supplementation of diets with exotic fruits positively affects plasma lipid profile and antioxidant activity in rats fed cholesterol-containing diets.

E0114 Expression of pregnancy-association plasma protein A and inducible nitric oxide synthase in the wall of balloon injured and early atherosclerotic porcine coronary artery

PurposeTo investigate the role of pregnancy-associated plasma protein A (PAPP-A), a novel marker of atherosclerotic plaque activity, in the progress of injured-restenosis and atherosclerosis, and the relationship between the expression...