Cholesterol-fed Group Research Articles

Exploration of Antidiabetic, Cholesterol-Lowering, and Anticancer Upshot of Probiotic Bacterium Pediococcus pentosaceus OBK05 Strain of Buttermilk.

Chronic metabolic disorders such as hyperglycemia (diabetes), hyperlipidemia (high cholesterol), and cancer have become catastrophic diseases worldwide. Accordingly, probiotic intervention is a new approach for alleviating catastrophic diseases. In the present study, Pediococcus pentosaceus OBK05 was investigated as a potential probiotic bacterium for antidiabetic, cholesterol-lowering, and anticancer activities by in vitro and in vivo studies. Cell-free supernatant (CFS) of OBK05 showed potent antidiabetic activity by inhibition of α-amylase (72 ± 0.9%) and α-glucosidase (61 ± 0.8%) activity in vitro when compared to that of acarbose as standard (86 ± 0.9%). Furthermore, the in vivo cholesterol-lowering activity of OBK05 was studied using cholesterol-fed hypercholesterolemic mice. When compared to the cholesterol-fed group, the OBK05-intervened cholesterol-fed mice group had significantly lowered the levels of lipids and showed significantly lower lipid droplet accumulation in the histology of hepatocytes. Similarly, CFS exhibited higher anticancer activity (87.57 ± 1.27%) against HT-29 cells with an IC50 of 54.51 ± 1.8. Fluorescence-activated cell sorting (FACS) analysis revealed that CFS induced the cell cycle arrest by inhibiting the G1 to S phase transition. Further, in vivo anticancer activities were confirmed in BALB/c mice models, which were divided into four groups and held for 16 weeks. HT-29 cells (5×109/mice) were injected subcutaneously twice (0 and 2nd week), and OBK05 (3 ×105 CFU/mL) was given orally to the respective groups. The OBK05-intervened HT-29 cell-induced mice group showed significant improvement at the histological level and alleviated the degree of atypia in the intestine.

Sweet Potato Leaf Feeding Decreases Cholesterol, Oxidative Stress and Thrombosis Formation in Syrian Hamsters with a High-Cholesterol Diet

Nutritional strategies to reduce hyperlipidemia and the risk of cardiovascular disease are gaining more public favor and medical professionals’ attention. The authors of this study explored the effect of sweet potato leaf powder (SPLP) feeding on the parameters of plasma lipids, reactive oxygen species, and time to thrombosis formation in Syrian hamsters fed with high-cholesterol diets. The animals were separated into six groups: a feeding control diet, a control diet containing 0.1% cholesterol, a control diet containing 0.2% cholesterol, a control diet containing 0.1% cholesterol plus 2.5% SPLP, a control diet containing 0.1% cholesterol plus 5% SPLP, and a control diet containing 0.2% cholesterol plus 5% SPLP for six weeks. The levels of serum total cholesterol (51% increase), low-density lipoprotein cholesterol (70.6% increase), very-low-density lipoprotein cholesterol (51.3% increase), and the triglyceride and atherogenic index (LDL-C/HDL-C) significantly increased in the high-cholesterol diet groups. Concomitant 5% sweet potato leaf powder ingestion significantly decreased the lipid profiles, with a 20.6% total cholesterol reduction in the 0.1% cholesterol diet groups, a 17.2% reduction in the 0.2% group, a 48.7% LDL reduction in the 0.1% cholesterol group, and a 30.3% reduction in the 0.2% group, with a consequent decrease in the atherogenic index. SPLP feeding was found to be associated with increased fecal sterol contents, with a 188.6% increase in the 0.1% cholesterol-fed group and a 177.3% increase in the 0.2% group. The SPLP-fed groups had depressed ROS levels, elongated FeCl3-induced times to thrombosis formation, and increased liver superoxide dismutase contents and SREBP-1 protein expression. Sweet potato leaf intake could reduce plasma total cholesterol, LDL, and oxidative stress. We suggest sweet potato leaf intake as a choice of nutritional strategy for hyperlipidemia and cardiovascular disease prevention.

RESTRAIN HYPERCHOLESTEROLEMIA WITH ORANGE AND MANDARIN VOLATILE AND FOLDED OILS

ABSTRACT The findings of this study proved that orange peels contain more twice oil yield over mandarin peels (0.24% and 0.12%, respectively). Concentrated orange and mandarin peel essential oils were obtained by fractional distillation. The chemical constituents of original orange and mandarin oils (OO and MO) and their fivefold (5FOO and 5FMO) were fractionated and identified by GC/MS and GC. Limonene was the major monoterpene component in orange and mandarin oils, 89.65% and 65.57%, respectively, followed by myrcene (3.95%) and γ-terpinene (23.07%) in orange and mandarin oils, respectively. Octanal (1.47%) and linalool (1.5%) were the abundant oxygenated components in orange and mandarin oils, respectively. The decrement percentages of limonene were 18.58 and19.25% in five fold orange and mandarin oils, respectively. The major oxygenated component in 5FOO was the alcoholic β- Cis-terpeneol (4.27%). The main esters of mandarin oil, methyl N-methyl anthranilate and geranyl acetate were increased to 0.45% and 1.05%, respectively in 5FMO. In general, total oxygenated compounds percentages increased by 6.9 and7.3 times after folding of orange and mandarin oils, respectively. The above mentioned oils, pure limonene and synthetic antioxidant (BHT) were orally administrated for hypercholesterolemic rats for four weeks. No significant differences were recorded among groups of rats administrated with all different tested oils in body weight gain (%) or feed intake (g) (p≤ 0.05). In general, all rat groups administrated with orange and mandarin oils and their concentrates, showed improvement in HDL levels nearly to normal level compared to the negative control. Thus, the decrement of serum cholesterol level among cholesterol-fed groups did not correlate with the amount of limonene consumed by rats and may be related to other minor components associated with limonene and shared in the antioxidant effect. The efficiency of folded oils on hypocholesterolemic rats did not affect by decreasing limonene by deterpenation process.

Changes in Intestinal Gene Expression of Zebrafish (Danio rerio) Related to Sterol Uptake and Excretion upon β-Sitosterol Administration

Replacement of fishmeal with plant ingredients will introduce not only plant oil and protein but also phytosterol to the fish diet. Mammals strictly restrict the uptake of phytosterol at intestinal epithelial cells by regulating the gene expressions of sterol uptake and excretion proteins; however, phytosterol is found in the fish muscle and other organs. In order to assess the ability of phytosterol uptake by the intestinal epithelial cells of fish, no-sterol diet, cholesterol-, and β-sitosterol-containing diet was separately administered to zebrafish, and the relative mRNA expressions related to sterol uptake and excretion were evaluated. Gene expression of Niemann-Pick C1-like protein 1 in the sitosterol-fed group was significantly higher than that of the cholesterol-fed group (p < 0.05). The expression of apolipoprotein A-I gene was also higher in the sitosterol-fed group than that in the no-sterol and cholesterol-fed groups. The expressions of ATP-binding cassette, sub-family G, member 5 and 8, were significantly higher in the sitosterol-fed group, compared to the no-sterol group. Regarding the gene expression of ATP-binding cassette sub-family A, member 1, the sitosterol-fed group showed higher expression level compared to the other groups (p < 0.01). These results suggest that fish should be tolerant to phytosterols in contrast to mammals.

Effects of garlic oil on interleukin-6 mediated cardiac hypertrophy in hypercholesterol-fed hamsters.

Hypercholesterol diets are the major causes of cardiac hypertrophy and various cardiac disorders. The purpose of this study is to evaluate the effects of garlic oil on cardiac hypertrophy induced by hypercholesterol diets. Golden Syrian hamsters were fed with 2% cholesterol or 2% cholesterol plus 1% garlic oil for 2 months. Heart architecture changes were measured by hematoxylin-eosin staining and the molecular mechanism was determined by western blotting. Garlic oil reduced whole-heart weight to bone weight ratio, and left ventricle weight to bone weight ratio in the cholesterol-fed group. Moreover, the garlic oil group showed significantly reduced interleukin-6, phosphorylated (p)-extracellular signal-regulated kinase-5, p-mitogen-activated protein kinase-5, calcineurin, nuclear transcription factor of nuclear factor of activated T-cells-3 and p-GATA binding protein 4 when compared with the cholesterol group. However, no changes were observed in gp-130, signal transducer and activator of transcription-3, p-P38 and p-Jun N-terminal kinases protein levels in all groups. The results show that garlic oil may be useful in the treatment of hypertrophy-associated cardiovascular diseases.

Para-Tyrosine Supplementation Improves Insulin- and Liraglutide- Induced Vasorelaxation in Cholesterol-Fed Rats.

Former data of our workgroup indicated that the accumulation of oxidized amino acids (meta- and ortho-tyrosine) due to oxidative stress may play an important role in the impaired insulininduced vasoactive properties of different arterial segments. There are evidences, that incorporation of these amino acids into cellular proteins leads to certain hormonal resistances, which might be restored by supplementation with the physiologic isoform, para-tyrosine. Rats in the control group were kept on a regular diet, rats in the cholesterol-fed group received high-fat diet, while the third group of rats received high-fat diet with para-tyrosine supplementation for 16 weeks. Plasma cholesterol level was significantly higher in the cholesterol-fed group, while the level of cholesterol in the cholesterol+para-tyrosine group did not differ significantly from that of the controls. Plasma level of insulin after glucose stimulation was decreased in the cholesterol-fed group, while that in the para-tyrosine supplemented group did not differ significantly from the controls. Vascular para-, meta- and ortho-tyrosine content was measured with HPLC. Elevated vascular meta-tyrosine/para-tyrosine ratio of cholesterol fed rats could be avoided by para-tyrosine supplementation. Vascular response of the thoracic aorta to insulin and liraglutide was assessed by a DMT multi-myograph. Cholesterol feeding resulted in vascular insulin-and liraglutide resistance, which was restored by para-tyrosine supplementation. Incorporation of the oxidative stress induced pathological tyrosine isoforms leads to vascular-hormone-resistances. We show that the physiological amino acid para-tyrosine is capable of restoring hypercholesterolemia-induced increased meta-tyrosine content of the vascular wall, thus attenuating functional vascular damage.

Anti-Oxidant and Anti-Inflammatory Effects of Vildagliptin in Non-Alcoholic Fatty Liver Disease of Mice

Background: Vildagliptin is one of the Dipeptidyl Peptidase-4 (DPP-4) inhibitors. It is used as a novel oral anti-diabetic drug for the treatment of Type 2 Diabetes Mellitus (T2DM) with significant control of glycated hemoglobin and a lower incidence of hypoglycemia or weight gain. Aim: The present study hypothesized that the DPP-4 inhibitor vildagliptin may exert protective effects on liver in Non-Alcoholic Fatty Liver Disease (NAFLD) in cholesterol-fed albino mice. Methods: Thirty six albino mice were divided into 3 group (n=12). Group 1 (control) received distilled water and fed normal diet. Group 2 fed high cholesterol diet without therapy. The third group was fed with cholesterol rich diet and daily treated with vildagliptin (15 mg/ kg/day for 12 weeks). Liver index, liver triglyceride contents, serum hepatic transaminases, some anti-oxidant enzymes and TNF-α levels were measured in all groups. Results: Administration of vildagliptin in cholesterol-fed mice attenuated the development of NAFLD via a significant (p<0.05) decrease in ALT & AST liver enzymes versus NAFLD non-treated group (33.43 ± 6.0 & 34.13 ± 4.7U/L versus 344.9 ± 12.1 & 278.9 ± 5.9 respectively). Additionally, both liver triglyceride contents and tumor necrosis factor-alpha (TNF-α) were significantly (p<0.05) reduced in treated group 3 compared to cholesterol-fed non-treated groups (for liver triglycerides: 12.61 ± 0.41 versus 56.41 ± 2.11 mg/g tissue protein respectively while for TNF-α: 1.25 ± 0.75 versus 10.78 ± 1.18pg/mL respectively). Meanwhile, it produced a significant (p<0.05) increase in the levels of measured serum antioxidant markers in group 3 versus group 2 as follow: (SOD enzyme: 37.5 ± 4.24 versus 0.84 ± 0.43IU/mL, TBARS: 3.46 ± 0.66 versus 62.5 ± 3.3nmol/mg hepatic tissue protein, Catalase enzyme 54.7 ± 3.45 versus 4.51 ± 0.32 and GPx enzyme 17.34 ± 0.79 versus 0.21 ± 0.02 unit/mg hepatic tissue protein, respectively). Conclusion: These results suggest possible anti-oxidant and partial anti-inflammatory mechanisms in therapy of NAFLD by this anti-diabetic drug.

Investigation of the Memory Impairment in Rats Fed with Oxidized-Cholesterol-Rich Diet Employing Passive Avoidance Test

Recent studies have shown that hypercholesterolemia, besides being a risk factor for cardiovascular diseases, has also toxic effects on central nervous system. The design of the present study was to investigate the effects of dietary cholesterol and oxidized cholesterol on cognitive function. Male Wistar rats were randomly divided into 3 groups. The animals were fed with three normal, 2% cholesterol-rich, and 2% oxidized cholesterol-rich diets for 14 weeks. Memory impairment was analyzed by passive avoidance test. Coenzyme Q10 content was also measured by a validate RP-HPLC method. Besides, lipid peroxidation in serum and brain tissue was determined by malondialdehyde concentration measurement. The results showed that feeding rats with high oxidized cholesterol diet for 14 weeks significantly impaired the cognitive function compared to the normal (P<0.001) and high cholesterol-fed groups (P<0.01). The memory impairment was positively correlated to the serum level of the oxidized LDL; it was significantly associated with the increased malondialdehyde concentration on the brain tissue of both groups (P<0.05 and P<0.001, respectively). The total antioxidant level in the serum was also decreased in rats fed with the oxidized cholesterol (P<0.05). Moreover, the brain coenzyme Q10 content was significantly declined in the animals fed with the oxidized cholesterol-rich diet compared to the animals fed with the normal (P<0.01) and cholesterol-rich diets (P<0.05). The results suggested that the high dietary intake of the oxidized-cholesterol might impair the memory that could be correlated to the oxidative stress and declined the coenzyme Q10 content of the brain tissue.

Dietary cholesterol and oxidised cholesterol: effects on sperm characteristics, antioxidant status and hormonal profile in rats

Present study was designed to compare the potential effects of high serum levels of LDL and oxidised LDL (OxLDL) on spermatogenesis parameters in male Wistar rats. Animals were allocated into three groups and were fed for 14weeks with normal, cholesterol-rich and oxidised cholesterol-rich diets. Blood lipid profile, sex hormones level, as well as sex organs weight were evaluated. The sex organs weight in oxidised cholesterol-fed group was significantly reduced (P<0.05). Spermatozoa count in the group with high serum concentration of OxLDL (64±4.2×10(6) ) was markedly lower (P<0.01) than that of normal rats (87±4.1×10(6) ) and rats with high serum level of LDL (90±6.3×10(6) ). Similarly, the percentage of viable spermatozoa was significantly (P<0.001) decreased from 78% to 52% by high level of OxLDL in serum. While, nonoxidised LDL did not have suppressive effects on spermatogenesis and organs weight. Consistent with these effects, the serum concentration of sex hormones including FSH (P<0.001), LH (P<0.001) and testosterone (P<0.01) was significantly decreased only in rats with high level of OxLDL but not in rats with high level of nonoxidised LDL. In conclusion, high OxLDL level showed higher destructive effect on reproductive system compared to the high LDL level.

Biochemical Alterations and Endothelial Dysfunction in Hypercholesterolemic Rats: Effects of Bezafibrate

Background: Hypercholesterolemia is a serious problem cause biohazards for human health and endothelial dysfunction. Objective: The current study was conducted to test effects of hyperlipidemia alone or in combination with bezafibrate at various doses. Total cholesterol, triacylglecerol, high density lipoprotein (HDL), low density lipoprotein (LDL), apolipoprotein B, endothelin-1, histamine, nitric oxide (NO), IL-6, creatinine and alanine amino transferase (ALT) were assayed Methodology: Rats were randomly divided into four groups (10 rats each). The first group fed a normal diet and represents the control group. The second group fed normal diet enriched with 1% cholesterol and 5% coconut oil (cholesterol-fed group). The third group fed on normal diet and bezafibrate at a dose of 100 mg/kg/day. The fourth group is cholesterol fed and subdivided into A, B, C and supplemented with bezafibrate at various concentration 50, 100, 200 mg/kg/day respectively. Feeding was continued daily for 6 weeks after incidence of hypercholesterolemia. Results: The current study revealed that total cholesterol, triacylglycerols, apolipoprotein B, and LDL concentration were significantly elevated in the cholesterol-fed rats compared to control alone and control plus bezafibrate. In addition, plasma endothelin-1, histamine and IL-6 concentration were significantly elevated in the cholesterol-fed rats compared to bezafibrate and control group rats. Bezafibrate administration to rats fed high cholesterol diet normalized significantly the changes in all parameters except NO levels compared to hyperlipidemic group. Meanwhile, the plasma nitric oxide in all groups was not significantly different from those of control. However, plasma HDL concentration in the cholesterol fed rats was significantly lowered compared to bezafibrate treated rats. Conclusion: Bezafibrate may acts as a mixed blessing drug to normalize lipid profiles and endothelial dysfunction during hyperlipidemia. © 2013 GESDAV

Low-Dose Fish Oil Consumption Prevents Hepatic Lipid Accumulation in High Cholesterol Diet Fed Mice

We examined the effects of low-dose fish oil ingestion on hepatic lipid accumulation caused after high cholesterol feeding in C57BL/6J mice. The mice were fed purified experimental diets consisting of 20 energy % (en%) safflower oil (SO or SO/CH), 2 en% fish oil + 18 en% safflower oil (2FO or 2FO/CH), or 5 en% fish oil + 15 en% safflower oil (5FO or 5FO/CH) with or without 2 weight % (wt %) cholesterol for 8 weeks. Hepatic triglyceride and total cholesterol contents were significantly lower in groups that were fed diets containing fish oil and cholesterol than in those that were fed safflower oil and cholesterol. The hepatic mRNA levels of fatty acid synthase (FAS) were lower in groups fed cholesterol or fish oil. Fatty acid oxidation-related hepatic gene expressions were higher in fish oil-fed groups. Fecal cholesterol excretion was higher in all cholesterol-fed groups; cholesterol excretion was high in groups fed fish oil and cholesterol. These results suggest that low-dose fish oil diets improve lipid metabolism by modifying the expression of lipid metabolism-related genes in the liver and increasing fecal cholesterol excretion.

Chemical composition and effect of an essential oil of Salix aegyptiaca L., Salicaceae, (musk willow) in hypercholesterolemic rabbit model

The essential oils (EO) of Salix aegyptiaca L., Salicaceae (SA), leaves were extracted using the hydrodistillation method and their chemical composition was further determined by GC-MS. 1,4-Dimethoxybenzene was the main isolated compound. Other major isolated constitutes were phenylethyl alcohol, carvone, citronellol, methyleugenol, eugenol, n-tetradecane and 4'-methoxyacetophenone. Twenty rabbits were equally divided into four groups: Normal control (NC) which fed a standard diet and three cholesterol-fed groups: HC, HC+1.0% SA and HC+3.0% SA groups which received 0.0%, 1.0% and 3.0% EO, respectively for four weeks. The serum lipid and lipoprotein profiles and atherogenic index (AI) were measured weekly. The high cholesterol diet significantly raised the TC, LDL-C, VLDL-C, HDL-C, TG and AI level compared with NC group. HC+1.0% SA and HC+3.0% SA groups showed similar results compared with HC group. It can be concluded that the EO of SA leaves could not prevent dyslipidemia that occurred in rabbits following inclusion of cholesterol in diet in both dose-and time-dependent manners.

Cholesterol diet leads to attenuation of ischemic preconditioning-induced cardiac protection: the role of connexin 43

Cardioprotection by ischemic preconditioning (IP) was abolished in connexin 43 (Cx43)-deficient mice due to loss of Cx43 located in mitochondria rather than at the sarcolemma. IP is lost in hyperlipidemic rat hearts as well. Since changes in mitochondrial Cx43 in hyperlipidemia have not yet been analyzed, we determined total and mitochondrial Cx43 levels in male Wistar rats fed a laboratory chow enriched with 2% cholesterol or normal chow for 12 wk. Hearts were isolated and perfused according to Langendorff. After a 10-min perfusion, myocardial tissue cholesterol, superoxide, and nitrotyrosine contents were measured and Cx43 content in whole heart homogenate and a mitochondrial fraction determined. In the cholesterol-fed group, tissue cholesterol and superoxide formation was increased (P < 0.05), while total Cx43 content remained unchanged. Mitochondrial total and dephosphorylated Cx43 content decreased. Hearts were subjected to an IP protocol (3 × 5 min ischemia-reperfusion) or time-matched aerobic perfusion followed by 30-min global ischemia and 5-min reperfusion. IP reduced infarct size in normal but not in cholesterol-fed rats. At 5-min reperfusion following 30-min global ischemia, the total and dephosphorylated mitochondrial Cx43 content was increased, which was abolished by IP in both normal and high-cholesterol diet. In conclusion, loss of cardioprotection by IP in hyperlipidemia is associated with a redistribution of both sarcolemmal and mitochondrial Cx43.

Effects of immunomodulatory drugs on plasma inflammatory markers in a rabbit model of atherosclerosis

Atherosclerosis is a chronic disease of the arterial wall where both innate and adaptive immuno-inflammatory mechanisms are involved. Inflammatory cytokines are implicated in the development and progression of atherosclerotic lesions. Immunomodulatory therapies have been proposed for the treatment of atherosclerosis. Therefore, the aim of this study was to investigate the systemic anti-inflammatory and immunomodulatory effects of atorvastatin, cyclosporine A (CsA), and tacrolimus (FK506) on plasma inflammatory markers in atherosclerotic rabbits. Male New Zealand rabbits were randomized into five groups each of 12 animals. Standard diet-fed group served as control, and the cholesterol-fed group received a diet supplemented with 1% cholesterol alone, cholesterol + atorvastatin, cholesterol + FK506, and cholesterol + CsA. Serum levels of lipid profile parameters (triglycerides, cholesterol, and high-density lipoprotein) were measured using colorimetric methods. Serum levels of C-reactive protein (CRP), interleukin-6 (Il-6), and interferon-gamma (INF-γ) were measured in all studied groups using ELISA techniques. Our results revealed a significant decrease (p < 0.001) in the serum levels of lipid profile parameters, CRP, Il-6, and INF-γ in atorvastatin-treated group compared with the cholesterol-fed group. On the other hand, a non-significant difference was observed for the same parameters in either FK506- or CsA-treated groups compared with the cholesterol-fed group. In conclusion, atorvastatin has a systemic anti-inflammatory role that far surpassed the cholesterol reduction effect alone. FK506 or CsA failed to suppress elevated plasma inflammatory markers. Thus, low doses of these two immunomodulating drugs could not have generalized systemic anti-inflammatory or immunosuppressive effects.

Neurovascular changes measured by time‐of‐flight MR angiography in cholesterol‐fed rabbits with cortical amyloid β‐peptide accumulation

To test the hypothesis that narrowing of cranial blood vessels in cholesterol-fed rabbits is a function of the duration of the high cholesterol diet. Such neurovascular changes, caused by elevated serum cholesterol, are linked to stroke and Alzheimer's disease risk. Four groups of New Zealand White rabbits were studied. Six were fed a normal diet, 19 were fed a 2% cholesterol diet with 0.12 ppm copper in the drinking water for 8 weeks, 10 weeks, or 12 weeks. Time-of-flight (TOF) MR angiography (MRA) at 3 Tesla was used to measure arterial diameters in 11 vessels. Previously published data for amyloid beta-peptide (Abeta) accumulation in the brains measured postmortem were correlated to vessel diameters. Ventricular volumes of rabbits were measured on group-averaged data. Several vessel diameters decreased with cholesterol diet duration. The posterior communicating arteries showed the largest significant effect. Abeta accumulation was inversely correlated with arterial diameter. Ventricular volumes between the normal diet and 12 weeks cholesterol-fed groups were not significantly different. Reduction in vessel diameter of medium-sized vessels but not large vessels was measured in these hypercholesterolemic rabbits. The vessel diameter narrowing and cortical Abeta deposition occurred before measurable ventricular enlargement.

Cholesterol diet-induced hyperlipidemia impairs the cardioprotective effect of postconditioning: role of peroxynitrite

The aim of the present study was to investigate if hyperlipidemia interferes with the infarct size-limiting effect of postconditioning and to study the involvement of peroxynitrite in this phenomenon. Rats were fed a 2% cholesterol-enriched or normal diet for 12 wk. Infarct size by triphenyltetrazolium chloride staining was measured in hearts isolated from both groups and subjected to 30 min coronary occlusion followed by 120 min reperfusion with or without the postconditioning protocol induced by six cycles of 10 s coronary occlusion and 10 s reperfusion at the onset of the reperfusion. Postconditioning significantly decreased infarct size in the normolipidemic but not in the hyperlipidemic group. Postconditioning increased cardiac 3-nitrotyrosine concentration (a marker for peroxynitrite formation) in the normal but not in the cholesterol-fed group when measured at the 5th min of reperfusion. Next, we tested if the postconditioning-induced acute increase in peroxynitrite is involved in the cardioprotection in normolipidemic animals in separate experiments. Postconditioning failed to decrease infarct size in the presence of the peroxynitrite decomposition catalyst 5,10,15,20-tetrakis-[4-sulfonatophenyl]-porphyrinato-iron [III] (20 mg/l) in normolipidemic animals. We conclude that an early increase in peroxynitrite after postconditioning plays a role in cardioprotection. Furthermore, hyperlipidemia blocks the cardioprotective effect of postconditioning at least in part via deterioration of the postconditioning-induced early increase in peroxynitrite formation.

Oral supplementation with l-aspartate and l-glutamate inhibits atherogenesis and fatty liver disease in cholesterol-fed rabbit

Previous studies have shown that dietary supplementation with L-aspartate and L-glutamate inhibits fatty streak initiation in cholesterol-fed rabbit. The present study investigates the role of dicarboxylic amino acids on the progression of fatty streaks and the development of fatty liver disease, which were caused in New Zealand White rabbits after a 0.5% w/w cholesterol diet for 7 weeks. A group of animals additionally received a combination of 12.5 mM L-aspartate and 12.5 mM L-glutamate per day through drinking water. Total cholesterol (TC), high-density lipoproteins cholesterol (HDLC), non-HDLC and triacylglycerol (TAG) concentrations were measured in plasma. Serum gamma-glutamyl transferase (gamma-GT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were also determined. At the end of dietary intervention, animals were sacrificed. Aortic, hepatic and brain lesions were evaluated after staining with hematoxylin and eosin. Supplementation with dicarboxylic amino acids inhibited the progression of aortic intima thickness (P < 0.05) and the development of liver lesions (P < 0.05). TC, non-HDLC and TAG were similarly increased in both cholesterol-fed groups. Serum gamma-GT and AST activities elevated during the study in all cholesterol-fed animals but the elevation of gamma-GT was milder and significantly lower in rabbits treated with L-aspartate and L-glutamate (P < 0.05). ALT activity was not affected by cholesterol feeding. In conclusion, oral supplementation with L-aspartate and L-glutamate inhibits the progression of atherogenesis and the development of fatty liver disease in the animal model of cholesterol-fed rabbit. The beneficial effects of dicarboxylic amino acids reflect the limited elevation of serum gamma-GT activity.

Effect of a cholesterol-rich diet on the metabolism of the free and esterified cholesterol components of a nanoemulsion that resembles LDL in rabbits

We have shown that the free cholesterol (FC) and the cholesteryl ester (CE) moieties of a nanoemulsion with lipidic structure resembling low-density lipoproteins show distinct metabolic fate in subjects and that this may be related to the presence of dyslipidemia and atherosclerosis. The question was raised whether induction of hyperlipidemia and atherosclerosis in rabbits would affect the metabolic behavior of the two cholesterol forms. Male New Zealand rabbits aged 4-5 months were allocated to a control group (N = 17) fed regular chow and to a 1% cholesterol-fed group (N = 13) during a 2-month period. Subsequently, the nanoemulsion labeled with 3H-FC and 14C-CE was injected intravenously for the determination of plasma kinetics and tissue uptake of the radioactive labels. In controls, FC and CE had similar plasma kinetics (fractional clearance rate, FCR = 0.234 +/- 0.056 and 0.170 +/- 0.038 h-1, respectively; P = 0.065). In cholesterol-fed rabbits, the clearance of both labels was delayed and, as a remarkable feature, FC-FCR (0.089 +/- 0.033 h-1) was considerably greater than CE-FCR (0.046 +/- 0.010 h-1; P = 0.026). In the liver, the major nanoemulsion uptake site, uptake of the labels was similar in control animals (FC = 0.2256 +/- 0.1475 and CE = 0.2135 +/- 0.1580%/g) but in cholesterol-fed animals FC uptake (0.0890 +/- 0.0319%/g) was greater than CE uptake (0.0595 +/- 0.0207%/g; P < 0.05). Therefore, whereas in controls, FC and CE have similar metabolism, the induction of dyslipidemia and atherosclerosis resulted in dissociation of the two forms of cholesterol.

The Effects of Splenectomy and Splenic Autotransplantation on Plasma Lipid Levels

Purpose: Atherosclerosis observations after splenectomy for trauma and hypersplenism suggests a possible role for the spleen in lipid metabolism. The authors examined the effects of splenectomy on serum lipids in rats and also cholesterol-fed rats with experimental atherosclerosis. Methods: This study was designed on rats. The rats were divided into five groups: splenectomy, normal diet (SP-N, n: 8), splenectomy, cholesterol-fed groups (SP-C, n: 8), splenic autotransplantation after splenectomy, normal diet (SA-N, n: 8), splenic autotransplantation after splenectomy, cholesterol-fed groups (SA-C, n: 8) and sham groups (n: 8). Total triglyceride, total cholesterol, HDL (high-density lipoprotein), LDL (low-density lipoprotein), and VLDL (very low-density lipoprotein) levels were determined in 40 rats. The rats were classified into five groups based on the surgical procedures. The spleens were removed and then the rats were fed a normal diet in Group SP-N (n = 8). The spleens were removed and then the rats were fed a diet containing 1% cholesterol in Group SP-C (n = 8). Splenectomy and splenic autotransplantations were performed and then the rats were fed a normal diet in Group SA-N (n = 8). Splenectomy and splenic autotransplantations were performed and then the rats were fed a diet containing 1% cholesterol in Group SA-C (n = 8). The rats were sham-operated in the control group (Group S, n = 8). An active splenic function was shown in rats that underwent splenic autotransplantation in both groups by using Technicium 99 m sulphurcolloide sintiscan on day 30. Blood lipid levels were repeated 6 months later. Results: There was no difference between pre- and postoperative lipid levels in the sham group and SA-N group (p >.05). All lipid levels including HDL were increased significantly in SP-C group (p <.05). Also VLDL and total tryglyceride levels were increased significantly in SP-N and SA-C groups (p <.05). Conclusions: This study showed that the spleen might have an important effect on lipid metabolism and splenic autotransplantation may be protective in conditions with increased lipid levels.

Effects of Feeding Periods of High Cholesterol and Saturated Fat Diet on Blood Biochemistry and Hydroxyproline Fractions in Rabbits

Hypercholesterolemia and hypertriglyceridemia are considered as important risk factors during the atherosclerotic process. The aim of the present investigation was to study the total cholesterol (TC), low-density lipoprotein cholesterol (LDLC), high density lipoprotein (HDL), triglyceride (TG), platelet levels and hydroxyproline fractions during the pathogenesis of atherosclerosis. For this purpose, twenty five 12-weeks, New Zealand white male rabbits, were purchased, individually caged, and divided into either control group or cholesterol-fed group. The control group (n = 10) was fed 100 g/day of normal diet, ORC-4 (Oriental Yeast Co. Ltd., Tokyo, Japan) for a period of 15 weeks. The cholesterol-fed group (n = 15) was fed a high cholesterol and saturated fat diet of ORC-4 containing 1% cholesterol plus 1% olive oil (100 g/day) for periods of 5 (group 1), 10 (group 2) and 15 (group 3) weeks. Blood sample from each animal was taken at the end of the experimental period for the biochemical analysis. The results of the present study showed that TC, LDLC, TG, HDLC and platelets were significantly (P < 0.01) increased in cholesterol-fed rabbits as compared with control rabbits. The serum hydroxyproline (Hyp) in rabbits belonging to group 1 showed no significant alteration when compared to control group. Group 2 rabbits showed a significant increase of 103% (P < 0.01) and 100% (P < 0.001) in free and protein-bound hydroxyproline fractions respectively when compared to control rabbits. However, there was no significant change in peptide-bound and total serum hydroxyproline levels as compared to the control group (P > 0.05). There was no significant (P > 0.05) decrease of free serum hydroxyproline in group 3 rabbits when compared to control rabbits. On the other hand, group 3 rabbits showed a significant increase in peptide-bound and protein-bound Hyp by 517% (P < 0.05) and 100% (P < 0.01) respectively when compared to control rabbits. However, total serum Hyp in group 3 rabbits showed no significant (P > 0.05) change when compared to control rabbits. These results suggest that feeding rabbits high cholesterol and saturated fat diet for feeding periods of 5, 10 and 15 weeks induced significant change in TC, LDLC, HDL, TG, platelet levels and various Hyp fractions in serum without any significant change in the total Hyp content.