Cholesterol-rich Diet Research Articles

Hypocholesterolemic effects of diets containing different levels of kishk as a dried fermented milk–whole wheat mixture in experimental rats

BackgroundKishk is a popular traditional functional food in Egypt. This study was performed to investigate the effects of different levels of kishk as a dried fermented milk/whole wheat mixture on growth performance, relative weight of organs, lipid profile, and some biochemical parameters in rats fed a cholesterol-rich diet. MethodsForty male rats were assigned to five groups, each consisting of eight rats. The first one presents the negative control group that received the basal diet, while the second group that serves as the positive (+) control group received a high-cholesterol diet (HCD). The last three groups received HCD supplemented with 10%, 20%, and 30% of kishk. ResultsRats fed diets containing various levels of kishk for 8 weeks had significantly (p < 0.05) lower body weights compared with the rats of both negative and positive groups. The liver/body weight ratio significantly increased in rats fed HCD compared with the control rats. Incorporation of kishk into the HCD at levels of 10%, 20%, and 30% significantly (p ≤ 0.05) decreased the change of liver/body weight ratio by 14.46%, 17.51%, and 18.78%, respectively, when compared with the HCD group. Results also indicate that rats fed HCD had a state of dyslipidemia, compared with the negative control group. Administration of HCD supplemented with various levels of kishk for 8 weeks significantly (p < 0.05) attenuated the increases in serum cholesterol, low-density lipoprotein cholesterol, triglyceride concentration, and atherogenic indices, and increased high-density lipoprotein cholesterol in a dose-dependent manner compared with the HCD group. Activities of liver enzymes (alanine transferase and aspartate transferase) as well as kidney function parameters (urea, uric acid, and creatinine) were elevated in the HCD group compared with the negative control group. ConclusionConsumption of HCD supplemented with various levels of kishk for 8 weeks induced a significant protective effect reflected in the reductions of the serum levels of aspartate transferase and alanine transferase, as well as kidney functions (uric acid, urea, and creatinine).

A Study to Assess the Effect of Cholesterol Rich Diet on Sputum Status in Patients with Pulmonary Tuberculosis Admitted in a Selected Hospital of Mumbai

Effect of cholesterol rich diet on sputum status in patients with pulmonary tuberculosis admitted in a selected hospital of Mumbai. Quasi experimental approach was used. Sample: patients diagnosed with tuberculosis with sputum positive status. Tool: Cholesterol rich diet, observation checklist and daily dietary inventory. Nineteen patients out of 30 from the sample group had sputum conversion from positive to negative within one month period. The critical ratio was calculated which was 5.54 which showed a significant difference between the percentages of study and control group. There was also improvement in the client's general wellbeing. The activity level of the clients was also improved a lot. The cholesterol rich diet though was not effective in improving the patients' hemoglobin levels, W.B.C count but it helped in early sputum conversion of the clients. The cholesterol which was supplemented to the clients did not raise clients' cholesterol levels above normal. Thus, it was concluded that cholesterol rich diet could help in early sputum conversion in clients with tuberculosis.The study guides the nurse to plan for such cholesterol rich dietary supplementation for the patients. Health education can be provided to nurses and patients about cholesterol rich dietary supplementation. This study also helps the nurses to know about importance in planning and providing nutrition.

Auto-inhibitory regulation of angiotensin II functionality in hamster aorta during the early phases of dyslipidemia

Emerging data point the crosstalk between dyslipidemia and renin-angiotensin system (RAS). Advanced dyslipidemia is described to induce RAS activation in the vasculature. However, the interplay between early dyslipidemia and the RAS remains unexplored. Knowing that hamsters and humans have a similar lipid profile, we investigated the effects of early and advanced dyslipidemia on angiotensin II-induced contraction. Cumulative concentration-response curves for angiotensin II (1.0pmol/l to 1.0µmol/l) were obtained in the hamster thoracic aorta. We also investigated the modulatory action of NAD(P)H oxidase on angiotensin II-induced contraction using ML171 (Nox-1 inhibitor, 0.5µmol/l) and VAS2870 (Nox-4 inhibitor, 5µmol/l). Early dyslipidemia was detected in hamsters treated with a cholesterol-rich diet for 15 days. Early dyslipidemia decreased the contraction induced by angiotensin II and the concentration of Nox-4-derived hydrogen peroxide. Advanced dyslipidemia, observed in hamsters treated with cholesterol-rich diet for 30 days, restored the contractile response induced by angiotensin II by compensatory mechanism that involves Nox-4-mediated oxidative stress. The hyporresponsiveness to angiotensin II may be an auto-inhibitory regulation of the angiotensinergic function during early dyslipidemia in an attempt to reduce the effects of the upregulation of the vascular RAS during the advanced stages of atherogenesis. The recovery of vascular angiotensin II functionality during the advanced phases of dyslipidemia is the result of the upregulation of redox-pro-inflammatory pathway that might be most likely involved in atherogenesis progression rather than in the recovery of vascular function. Taken together, our findings show the early phase of dyslipidemia may be the most favorable moment for effective atheroprotective therapeutic interventions.

Inhibition of extracellular matrix production and remodeling by doxycycline in smooth muscle cells

Alterations in the extracellular matrix (ECM) production and remodeling of smooth muscle cells (SMCs) have been implicated in processes related to the differentiation in atherosclerosis. Due to the anti-atherosclerotic properties of the tetracyclines, we aimed to investigate whether cholesterol supplementation changes the effect of doxycycline over the ECM proteins synthesis and whether isoprenylated proteins and Rho A protein activation are affected. SMC primary culture isolated from chicks exposed to atherogenic factors in vivo (a cholesterol-rich diet, SMC-Ch), comparing it with control cultures isolated after a standard diet (SMC-C). After treatment with 20 nM doxycycline, [H3]-proline and [H3]-mevalonate incorporation were used to measure the synthesis of collagen and isoprenylated proteins, respectively. Real-time PCR was assessed to determine col1a2, col2a1, col3a1, fibronectin, and mmp2 gene expression and the pull-down technique was applied to determine the Rho A activation state. A higher synthesis of collagens and isoprenylated proteins in SMC-Ch than in SMC-C was determined showing that doxycycline inhibits ECM production and remodeling in both SMC types of cultures. Moreover, preliminary results about the effect of doxycycline on protein isoprenylation and Rho A protein activation led us to discuss the possibility that membrane G-protein activation pathways could mediate the molecular mechanism.

Modulating the Gut Microbiota Improves Glucose Tolerance, Lipoprotein Profile and Atherosclerotic Plaque Development in ApoE-Deficient Mice.

The importance of the gut microbiota (GM) in disease development has recently received increased attention, and numerous approaches have been made to better understand this important interplay. For example, metabolites derived from the GM have been shown to promote atherosclerosis, the underlying cause of cardiovascular disease (CVD), and to increase CVD risk factors. Popular interest in the role of the intestine in a variety of disease states has now resulted in a significant proportion of individuals without coeliac disease switching to gluten-free diets. The effect of gluten-free diets on atherosclerosis and cardiovascular risk factors is largely unknown. We therefore investigated the effect of a gluten-free high-fat cholesterol-rich diet, as compared to the same diet in which the gluten peptide gliadin had been added back, on atherosclerosis and several cardiovascular risk factors in apolipoprotein E-deficient (Apoe-/-) mice. The gluten-free diet transiently altered GM composition in these mice, as compared to the gliadin-supplemented diet, but did not alter body weights, glucose tolerance, insulin levels, plasma lipids, or atherosclerosis. In parallel, other Apoe-/- mice fed the same diets were treated with ampicillin, a broad-spectrum antibiotic known to affect GM composition. Ampicillin-treatment had a marked and sustained effect on GM composition, as expected. Furthermore, although ampicillin-treated mice were slightly heavier than controls, ampicillin-treatment transiently improved glucose tolerance both in the absence or presence of gliadin, reduced plasma LDL and VLDL cholesterol levels, and reduced aortic atherosclerotic lesion area. These results demonstrate that a gluten-free diet does not seem to have beneficial effects on atherosclerosis or several CVD risk factors in this mouse model, but that sustained alteration of GM composition with a broad-spectrum antibiotic has beneficial effects on CVD risk factors and atherosclerosis. These findings support the concept that altering the microbiota might provide novel treatment strategies for CVD.

Proinflammatory Stimulation of Toll-Like Receptor 9 with High Dose CpG ODN 1826 Impairs Endothelial Regeneration and Promotes Atherosclerosis in Mice.

BackgroundToll-like receptors (TLR) of the innate immune system have been closely linked with the development of atherosclerotic lesions. TLR9 is activated by unmethylated CpG motifs within ssDNA, but also by CpG motifs in nucleic acids released during vascular apoptosis and necrosis. The role of TLR9 in vascular disease remains controversial and we sought to investigate the effects of a proinflammatory TLR9 stimulation in mice.Methods and FindingsTLR9-stimulation with high dose CpG ODN at concentrations between 6.25nM to 30nM induced a significant proinflammatory cytokine response in mice. This was associated with impaired reendothelialization upon acute denudation of the carotid and increased numbers of circulating endothelial microparticles, as a marker for amplified endothelial damage. Chronic TLR9 agonism in apolipoprotein E-deficient (ApoE-/-) mice fed a cholesterol-rich diet increased aortic production of reactive oxygen species, the number of circulating endothelial microparticles, circulating sca-1/flk-1 positive cells, and most importantly augmented atherosclerotic plaque formation when compared to vehicle treated animals. Importantly, high concentrations of CpG ODN are required for these proatherogenic effects.ConclusionsSystemic stimulation of TLR9 with high dose CpG ODN impaired reendothelialization upon acute vascular injury and increased atherosclerotic plaque development in ApoE-/- mice. Further studies are necessary to fully decipher the contradictory finding of TLR9 agonism in vascular biology.

Myeloid-Specific Deletion of Diacylglycerol Lipase α Inhibits Atherogenesis in ApoE-Deficient Mice.

BackgroundThe endocannabinoid 2-arachidonoylglycerol (2-AG) is a known modulator of inflammation. Despite its high concentration in vascular tissue, the role of 2-AG in atherogenesis has not yet been examined.MethodsApoE-deficient mice were sublethally irradiated and reconstituted with bone marrow from mice with a myeloid-specific knockout of the 2-AG synthesising enzyme diacylglycerol lipase α (Dagla) or control bone marrow with an intact 2-AG biosynthesis. After a cholesterol-rich diet for 8 weeks, plaque size and plaque morphology were examined in chimeric mice. Circulating inflammatory cells were assessed by flow cytometry. Aortic tissue and plasma levels of endocannabinoids were measured using liquid chromatography-multiple reaction monitoring.ResultsMice with Dagla-deficient bone marrow and circulating myeloid cells showed a significantly reduced plaque burden compared to controls. The reduction in plaque size was accompanied by a significantly diminished accumulation of both neutrophil granulocytes and macrophages in atherosclerotic lesions of Dagla-deficient mice. Moreover, CB2 expression and the amount of oxidised LDL within atherosclerotic lesions was significantly reduced. FACS analyses revealed that levels of circulating inflammatory cells were unaltered in Dagla-deficient mice.ConclusionsMyeloid synthesis of the endocannabinoid 2-AG appears to promote vascular inflammation and atherogenesis. Thus, myeloid-specific disruption of 2-AG synthesis may represent a potential novel therapeutic strategy against atherosclerosis.

Role of the ABC transporter Mdr49 in Hedgehog signaling and germ cell migration.

Coalescence of the embryonic gonad in Drosophila melanogaster requires directed migration of primordial germ cells (PGCs) towards somatic gonadal precursor cells (SGPs). It was recently proposed that the ATP-binding cassette (ABC) transporter Mdr49 functions in the embryonic mesoderm to facilitate the transmission of the PGC attractant from the SGPs; however, the precise molecular identity of the Mdr49-dependent guidance signal remained elusive. Employing the loss- and gain-of-function strategies, we show that Mdr49 is a component of the Hedgehog (hh) pathway and it potentiates the signaling activity. This function is direct because in Mdr49 mutant embryos the Hh ligand is inappropriately sequestered in the hh-expressing cells. Our data also suggest that the role of Mdr49 is to provide cholesterol for the correct processing of the Hh precursor protein. Supporting this conclusion, PGC migration defects in Mdr49 embryos are substantially ameliorated by a cholesterol-rich diet.

Effect of Morinda citrifolia (Noni) Fruit Juice on High Fat Diet Induced Dyslipidemia in Rats.

The medicinal value of Morinda citrifolia L. (commonly known as Noni) has been explored in ancient folk remedies with a wide range of therapeutic utility, including antibacterial, antiviral, antifungal, antitumour, analgesic, hypotensive, anti-inflammatory and immune enhancing effects. The present study was designed to evaluate the effects of Noni fruit juice on serum lipid profile in high fat diet induced murine model of dyslipidemia. Hyperlipidemia was induced by feeding a cholesterol rich high fat diet for 45 days in wistar albino rats of either sex (n=8). Noni fruit juice administered at 50mg/kg/day and 100mg/kg/day, per oral, was compared with the standard drug Atorvastatin (10mg/kg/day, oral) fed for the latter 30 days. The blood samples were then sent for complete blood lipid profile, after 30 days of treatment. The data presented as mean ± SEM was analyzed using one-way ANOVA followed by Tukey's post-hoc test. The p <0.05 was considered as statistically significant. The Noni fruit juice treated group showed a significant decrease in the total cholesterol, triglycerides and very low density lipoprotein - Cholesterol at both the doses when compared to the disease control (p<0.05). However, the decrease in the TC (102.75±9.79 mg/dL) and LDL-C (47.87±7.47 mg/dL) levels observed with the noni fruit juice at the 50mg/kg dose employed, failed to show a statistical significance when compared to atorvastatin. The present study provides evidence for the hypolipidemic activity of Noni fruit juice in high fat diet induced hyperlipidemia in rats.

Aktivitas Hipolipidemik dan Indeks Aterogenik yang Rendah Ekstrak Air Daun Tapak Dara pada Tikus Hiperkolesterolemia

Catharanthus roseus is one plant recognized as medical potential which can decrease cholesterol. Thepresent study was carried out to evaluate the effect of aqueous leaf extracts of C. roseus on plasma lipidprofil levels in rats cholesterol rich diet and atherogenic index. This study was carried out on 15 SpraqueDawley male rats randomly distributed into five groups (n=3). Rats hypercholesterolemic wereadministrated cholesterol rich diet containing 1% (w/w). Normal control group with normal diet whitoutextracts (K1), positive control hypercholesterolemic group (K2) with cholesterol rich diet whitout extracts,and others three groups (K3-K5) were feed high cholesterol and 1 mL of aqueous leaf extracts of C. roseuswith a dose of 20% (w/v), 40% (w/v) and 80% (w/v) respectively, administered twice daily 1 mL orally.Treatment was given for 28 days. After treatment, the plasma lipids profile such cholesterol total, highdensity lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TGA) level were measured. Theresults showed that during the period of treatment with cholesterol rich diet have produced under condition hypercholesterolemic of rats with average cholesterol levels of 124.33 ± 4.04 mg/dL (K2 group). Treatmentwith extract dose of 80% showed the best results among the other doses in reducing levels of total cholesterol,triglycerides, LDL and raise HDL levels. Extract treatment dose of 80% up to four weeks resulted incholesterol total (79.33±3.51 mg/dL), TGA (72.33±6.65 mg/dL), HDL (55.00±3.60 mg/dL), and LDL(9.87±5.34mg/dL). In addition, value ratio of cholesterol:HDL was 1.4: 1 and atherogenic index value was0.44. These results suggest that extracts of leaf C. roseus optimum doses 80% (w/v) has hipolipidemicactivity in hypercholesterolemic rats and it has low atherogenic index value.

High LDL levels lead to increased synovial inflammation and accelerated ectopic bone formation during experimental osteoarthritis

A relation between osteoarthritis (OA) and increased cholesterol levels is apparent. In the present study we investigate OA pathology in apolipoprotein E (ApoE)(-)(/-) mice with and without a cholesterol-rich diet, a model for high systemic low density lipoprotein (LDL) cholesterol levels independent of weight. Wild type (WT), Apoe(-)(/-), S100a9(-/-) and Apoe(-)(/-)S100a9(-/-) mice (C57BL/6 background) received a standard or cholesterol-rich diet. Experimental OA was induced by intra-articular injection of collagenase and animals were sacrificed at day 10 and day 36. Although minimal differences in cartilage damage were found between the WT and ApoE(-)(/-) mice, increased synovial thickening was found in the latter. Thirty-six days after OA-induction, ApoE(-)(/-) mice on a standard diet showed increased ectopic bone formation, particularly at the medial collateral ligament, compared with OA in WT mice. Furthermore, a significant increase in synovial gene expression of both S100a8 and S100a9 and S100A8/S100A9 protein levels was found in ApoE(-)(/-) mice, suggesting an activated inflammatory status of synovial cells. In both ApoE(-)(/-) and WT mice, addition of a cholesterol-rich diet resulted in excessive bone formation in the medial collateral ligament at late-time-point OA. Interestingly, at the early time point, proteoglycan deposition was already significantly increased in ApoE(-)(/-) mice compared with WT mice. Mice deficient for both ApoE and S100a9 also showed increased ectopic bone formation, but not synovial activation, suggesting a role for S100-proteins in cholesterol-mediated synovial activation. Increased cholesterol levels strongly elevate synovial activation and ectopic bone formation in early-stage collagenase-induced OA.

Cholesterol lowering effect in the gall bladder of dogs by Herniaria hirsuta

Infusions of Herniaria hirsuta L., H. glabra L. and H. fontanesii J. Gay (Caryophyllaceae) are well known in Moroccon folk medicine for the treatment of biliary dyskinesia, (uro)lithiasis or as a diuretic, and in Europe as an urological drug [1, 2]. An in vivo experiment to evaluate the cholesterol lowering effect of a decoction of H. hirsuta in the gall bladder of dogs was carried out. Three groups of dogs i.e. control dogs (CG), dogs treated with ursodeoxycholic acid (UDCA) (2 × 7.35 mg/kg body weight/day) and dogs treated with the standardized decoction (HG) (2 × 48.5 mg/kg body weight/day) were fed a fatty diet during 120 days after which a diet without additional fat was introduced till day 180 [3]. Treatment started 30 days after the start of the fatty diet and lasted until the end of the experiment. A bile and blood sample of each dog was collected every 30 days, after which the cholesterol level was determined. The in vivo experiments already showed a minor difference for bile cholesterol between CG and HG after 30 days of treatment with the decoction, and a more pronounced difference after 90 days of treatment. Even 30 days after discontinuation of the cholesterol-rich diet a significant difference remained between CG and HG. There was no significant difference in blood cholesterol between the groups. Prolonged use of this standardized H. hirsuta extract resulted in a cholesterol-lowering effect in the bile of dogs. Since this pharmacological effect prevents the formation of gallstones and can contribute to solving existing gallstones, a standardized decoction of H. hirsuta may have a positive effect in the treatment of gallstones in human patients.

Anti-atherosclerotic effects of garlic preparation in freeze injury model of atherosclerosis in cholesterol-fed rabbits

BackgroundGarlic (Allium sativum L.) is one of the most popular substances used to reduce various risks associated with cardiovascular disease. However, little is known on the direct effects of garlic on atherosclerosis. PurposeIn the present study we have examined the effect of per oral administration of the time-released garlic herbal preparation on serum atherogenicity and formation of intimal thickening after freeze injury in cholesterol-fed rabbits. MethodsGroup 1 rabbits maintained on the standard cholesterol-rich diet served as the control. Group 2 rabbits were fed the cholesterol-rich diet and treated with garlic preparation containing 300 mg garlic powder. ResultsLocal thickening of the aortic media (i.e., the neointima formation) in the freeze injury zone was observed in all the rabbits. Regular garlic preparation therapy prevented the neointima formation and the accumulation of free and esterified cholesterol, triglycerides, phospholipids and collagen in the neointima, the effects being statistically significant. Garlic preparation also decreased serum lipid content by 1.5-fold and lowered atherogenic activity of blood serum (ability to induce lipid accumulation in cultured cells) induced by cholesterol-rich diet. ConclusionThe results obtained indicate that garlic preparation prevents the development of cholesterol-induced experimental atherosclerosis and possesses the direct anti-atherogenic activity.

A Diet-Sensitive BAF60a-Mediated Pathway Links Hepatic Bile Acid Metabolism to Cholesterol Absorption and Atherosclerosis

Dietary nutrients interact with gene networks to orchestrate adaptive responses during metabolic stress. Here, we identify Baf60a as a diet-sensitive subunit of the SWI/SNF chromatin-remodeling complexes in the mouse liver that links the consumption of fat- and cholesterol-rich diet to elevated plasma cholesterol levels. Baf60a expression was elevated in the liver following feeding with a western diet. Hepatocyte-specific inactivation of Baf60a reduced bile acid production and cholesterol absorption, and attenuated diet-induced hypercholesterolemia and atherosclerosis in mice. Baf60a stimulates expression of genes involved in bile acid synthesis, modification, and transport through a CAR/Baf60a feedforward regulatory loop. Baf60a is required for the recruitment of the SWI/SNF chromatin-remodeling complexes to facilitate an activating epigenetic switch on target genes. These studies elucidate a regulatory pathway that mediates the hyperlipidemic and atherogenic effects of western diet consumption.

Thoracic Duct Narrowing-Innovative Technique Restraining Weight Gain in Rats.

The lymphatic system is responsible for the absorption of fats from the digestive system, conveying 60-70 % of ingested fat to the blood stream. From the anatomical point of view, all the lymphatic drainage from the lower half of the body converges in the abdomen to enter the thoracic duct. This experimental study aim was to study the result of thoracic duct narrowing (TDN), an innovative surgical technique, on weight gain restrain in high-fat diet-fed rats. Forty-seven rats were allocated into three groups: thoracic duct narrowing ("S"-surgery), sham operation ("CS"-control surgery), and no surgery ("C"-control). All rats were fed with high-fat, cholesterol-rich diet. Food consumption and metabolic syndrome parameters including weight gain, plasma lipids and glucose, blood pressure, and viscera weight and histopathology were analyzed. Thoracic duct narrowing was proved simple and safe surgical procedure in the rat model. TDN induced weight gain restrain, associated with mild hepatic steatosis compared to moderate-severe hepatic steatosis in control groups. Splenomegaly and splenic fatty histiocytes were shown in the treated animals. TDN improved several parameters of the metabolic syndrome in high-fat diet-fed rats. TDN carries the potential of innovative obesity treatment using the lymphatic route of lipid absorption.

Real-Time Elastography Visualization and Histopathological Characterization of Rabbit Atherosclerotic Carotid Arteries

To evaluate the feasibility of non-invasive vascular real-time elastography imaging (RTE) in visualizing the composition of rabbit carotid atherosclerotic plaque as determined by histopathology, a rabbit model of accelerated carotid atherosclerosis was used. Thirty rabbits were randomly divided into two groups of 15 rabbits each. The first group was fed a cholesterol-rich diet and received balloon-induced injury the left common carotid artery endothelium, whereas the second group only received a cholesterol-rich diet. The rabbits were all examined in vivo with HITACHI non-invasive vascular real-time elastography (Hi-RTE) at baseline and 12 wk, and results from the elastography were compared with American Heart Association histologic classifications. Hi-RTE and the American Heart Association histologic classifications had good agreement, with weighted Cohen's kappa (95% confidence internal) of 0.785 (0.649–0.920). Strains of segmented plaques that were stained in different colors were statistically different (p < 0.0001). The sensitivity and specificity of elastograms for detecting a lipid core were 95.5% and 61.5%, respectively, and the area under the receiver operating characteristic curve was 0.789, with a 95% confidence interval of 0.679 to 0.876. This study is the first to indicate the feasibility of utilizing Hi-RTE in visualizing normal and atherosclerotic rabbit carotid arteries non-invasively. This affordable and reliable method can be widely applied in research of both animal and human peripheral artery atherosclerosis.

A Hamster Model of Diet-Induced Obesity for Preclinical Evaluation of Anti-Obesity, Anti-Diabetic and Lipid Modulating Agents.

AimUnlike rats and mice, hamsters develop hypercholesterolemia, and hypertriglyceridemia when fed a cholesterol-rich diet. Because hyperlipidemia is a hallmark of human obesity, we aimed to develop and characterize a novel diet-induced obesity (DIO) and hypercholesterolemia Golden Syrian hamster model.Methods and ResultsHamsters fed a highly palatable fat- and sugar-rich diet (HPFS) for 12 weeks showed significant body weight gain, body fat accumulation and impaired glucose tolerance. Cholesterol supplementation to the diet evoked additional hypercholesterolemia. Chronic treatment with the GLP-1 analogue, liraglutide (0.2 mg/kg, SC, BID, 27 days), normalized body weight and glucose tolerance, and lowered blood lipids in the DIO-hamster. The dipeptidyl peptidase-4 (DPP-4) inhibitor, linagliptin (3.0 mg/kg, PO, QD) also improved glucose tolerance. Treatment with peptide YY3-36 (PYY3-36, 1.0 mg/kg/day) or neuromedin U (NMU, 1.5 mg/kg/day), continuously infused via a subcutaneous osmotic minipump for 14 days, reduced body weight and energy intake and changed food preference from HPFS diet towards chow. Co-treatment with liraglutide and PYY3-36 evoked a pronounced synergistic decrease in body weight and food intake with no lower plateau established. Treatment with the cholesterol uptake inhibitor ezetimibe (10 mg/kg, PO, QD) for 14 days lowered plasma total cholesterol with a more marked reduction of LDL levels, as compared to HDL, indicating additional sensitivity to cholesterol modulating drugs in the hyperlipidemic DIO-hamster. In conclusion, the features of combined obesity, impaired glucose tolerance and hypercholesterolemia in the DIO-hamster make this animal model useful for preclinical evaluation of novel anti-obesity, anti-diabetic and lipid modulating agents.

Protective effect of kombucha on rats fed a hypercholesterolemic diet is mediated by its antioxidant activity

Context: Kombucha (KT) is claimed to have various beneficial effects on human health, but there is very little scientific evidence available in the literature.Objective: The present study investigates the effects of Camellia sinensis (GT) Linn. (Theaceae) and KT, two natural drinks, on cholesterol and antioxidant status using a hypercholesterolemia rat model.Materials and methods: The present study compared the free-radical scavenging abilities and polyphenol levels of GT and KT. Wistar rats fed cholesterol-rich diets were given KT or GT (5 mL/kg body weight per day, po) for 16 weeks, then fasted overnight and sacrificed. The plasma lipid levels, thiobarbituric acid reactive substances (TBARS) and aspartate aminotransferase (AST), alanine aminotransferase (ALT), and γ-glutamyl transpeptidase (GGT) serum levels, antioxidant activities of superoxide dismutase (SOD) and catalase (CAT), and creatinine and urea rats were examined.Results: KT had a phenolic compound of 955 ± 0.75 mg GAE/g) followed, by GT (788.92 ± 0.02 mg GAE/g). The free radical scavenging activity of KT was higher than GT. Compared with GT, KT induced lowered serum levels of TC, TG, VLDL-C, and LDL-C by 26, 27, 28, and 36%, respectively, and increased the serum level of high-density lipoprotein cholesterol (HDL-C). KT induced a 55% decrease of TBARS level in liver and 44% in kidney, compared with those of rats fed a cholesterol-rich diet alone. Moreover, CAT and SOD activities were reduced by 29 and 33%, respectively, in liver and 31 and 35%, respectively, in kidney, after oral administration of KT, compared with those of HCD-fed rats.Conclusion: The findings revealed that KT administration induced attractive curative effects on hypercholesterolemic, particularly in terms of liver–kidney functions in rats. Its effect on humans needs to be studied further.

Abstract 554: Vascular Lesion Formation is Attenuated by Genetic Deletion of Hyaluronan Synthase 3

An extensive accumulation of the matrix component hyaluronan (HA) has been observed in vascular lesions. HA is thought to confer immunomodulatory actions and to regulate cell proliferation and migration in diseased vessels. HA is extruded into the extracellular space by three transmembrane HA-synthase isoenzymes (HAS1, -2, -3). Objective of the current study was to assess specifically the role of HAS3 in vascular pathologies such as restenosis and atherosclerosis. In HAS3 deficient mice (HAS3-/-) and wildtype controls (WT) neointimal hyperplasia was induced using a carotid artery ligation model. Morphometric analysis of carotid arteries harvested 28 days after ligation revealed that lack of HAS3 mitigates neointimal lesion formation as indicated by a decrease in intimal area (AUC: HAS3-/-, 1.53 x107 ± 0.23 x107 μm3 vs. WT, 2.59 x 107 ± 0.34 107 μm3, n = 9, p &lt; 0.05) and reduced intima/media ratio (HAS3-/-, 0.94 ± 0.13 vs. WT, 1.39 ± 0.13, n = 9, p &lt; 0.05). Supportive in vitro studies suggest that blunted neointimal hyperplasia is mainly due to a reduced migration and to a lesser extent reduced proliferation of vascular smooth muscle cell (VSMC). Next atherosclerosis-prone apolipoprotein E/HAS3 double-deficient (apoE-/-/HAS3-/-) mice were generated and fed with a cholesterol-rich Western type diet for 15 weeks. Analysis of aortic en face preparations stained with Oil Red O revealed reduced plaque burden compared to apoE-/- controls (apoE-/-/HAS3-/-, 6.95 ± 2.66, n = 10 vs. apoE-/-, 11.43 ± 4.06, n = 13, p &lt; 0.05). Plasma lipids (total cholesterol, LDL/VLDL, HDL cholesterol) did not differ between the groups. Plaque composition at the aortic root showed decreased collagen content (Picro Sirius Red staining) but no differences in cell density, VSMC content (alpha Actin staining) and macrophage count (Mac2 staining). In contrast, flow cytometry data point to a reduction in circulating T cells (apoE-/-/HAS3-/-, 28.4 ± 1.2 %, n = 5 vs. control, 41.7 ± 5.1 %, n = 6). Collectively, these findings suggest a role for HAS3 in vascular lesion formation. In neointimal hyperplasia HAS3-dependent HA synthesis may support mainly VSMC migration whereas in atherogenesis additional immunomodulatory actions are proposed.

Abstract 349: Sgk1 Is a Pivotal Player in Vascular Inflammation and Atherogenesis

Objective: Atherosclerosis, an inflammatory disease of arterial vessel wall, requires migration and MMP-9-dependent invasion of macrophages into the vascular wall. MMP-9 expression is stimulated by transcription factor NF-κB, which is regulated by IκB and thus IκB-kinase IKK. Regulators of NF-κB include SGK1. The present study explored involvement of SGK1 in vascular inflammation and atherogenesis. Approach and Results: Gene-targeted ApoE-deficient mice without (apoe-/-sgk1+/+) or with (apoe-/-sgk1-/-) additional SGK1 knockout received 16-weeks cholesterol-rich diet. Atherosclerotic lesions in aorta and carotid artery, vascular CD45+ leukocyte infiltration, Mac-3+ macrophage infiltration, VSMC content, MMP-2 and MMP-9 positive areas in atherosclerotic tissue were significantly less in apoe-/-sgk1-/-mice. Migration of SGK1-deficient CD11b+F4/80+ macrophages was significantly diminished in vitro and in vivo. Zymographic MMP-2 and MMP-9 production, MMP-9 activity and invasion through matrigel in vitro were significantly less in sgk1-/- than in sgk1+/+ macrophages and in control plasmid- or inactive K127NSGK1-transfected than in constitutively active S422DSGK1-transfected THP-1 cells. Plaques of apoe-/-sgk1-/- mice displayed reduced macrophage number and macrophage MMP-9 content. In THP-1 cells MMP-inhibitor GM6001 abrogated S422DSGK1-induced MMP-9 production and invasion. MMP-9 transcript levels were significantly reduced in sgk1-/- macrophages and strongly upregulated in S422DSGK1-transfected THP-1 cells compared to control plasmid- or K127NSGK1-transfected THP-1 cells. Phosphorylation of IKK and IκB and nuclear translocation of p50 were significantly lower in sgk1-/- macrophages than in sgk1+/+ macrophages and significantly higher in S422DSGK1-transfected THP-1 cells than in control plasmid- or K127NSGK1-transfected THP-1 cells. Treatment of S422DSGK1-transfected THP-1 cells with IKK-inhibitor BMS-345541 abolished S422DSGK1-induced increase of MMP-9 transcription and gelatinase activity. Conclusions: SGK1 plays a pivotal role in vascular inflammation during atherogenesis. SGK1 participates in the regulation of macrophage migration and MMP-9 transcription via regulation of NF-κB.