Effects of Cyclo-His-Pro-enriched yeast hydrolysate on blood glucose levels and lipid metabolism in obese diabetic ob/ob mice

BACKGROUND/OBJECTIVES

We examined the hypoglycemic and anti-hyperlipidemic effect of yeast hydrolysate (YH) enriched with Cyclo-His-Pro (CHP) in the C57BL/6J ob/ob mouse model.

MATERIALS/METHODS

Mice were separated into 4 groups (8 mice/group) on the basis of blood glucose and body weight: WT control, lean mice given vehicle; ob/ob control, ob/ob mice given vehicle; YH-1, ob/ob mice given 0.5 g/kg of YH; YH-2, ob/ob mice given 1 g/kg of YH. YH in saline or vehicle was administered orally in the same volume every day for 3 weeks.

RESULTS

Mice treated with YH (0.5 and 1 g/kg) for 3 weeks displayed a significant reduction in overall body weight gain and perirenal and epididymal adipose tissue weight compared to the ob/ob control group. Additionally, high-density lipoprotein (HDL) cholesterol, glucose, and atherogenic indexes were significantly decreased in the blood of YH-1 and YH-2 groups compared to the ob/ob control. In ob/ob mice, YH administration significantly improved glucose tolerance and blood insulin levels. These data indicate that YH treatment produces potent hypoglycemic and anti-hyperlipidemic effects by controlling body weight, fat mass, blood lipid, insulin levels, and glucose tolerance.

CONCLUSION

YH could potentially be used as a treatment option for diabetes and hyperlipidemia. The CHP-enriched YH may be a promising strategy in the development of hypoglycemic peptide nutraceuticals.     

Leptin deficiency-induced obesity affects the density of mast cells in abdominal fat depots and lymph nodes in mice

Background  

Mast cells are implicated in the pathogenesis of obesity and insulin resistance. Here, we explored the effects of leptin deficiency-induced obesity on the density of mast cells in metabolic (abdominal fat depots, skeletal muscle, and liver) and lymphatic (abdominal lymph nodes, spleen, and thymus) organs. Fourteen-week-old male leptin-deficient ob/ob mice and their controls fed a standard chow were studied. Tissue sections were stained with toluidine blue to determine the density of mast cells. CD117/c-kit protein expression analysis was also carried out. Furthermore, mast cells containing immunoreactive tumor necrosis factor-α (TNF-α), a proinflammatory cytokine involved in obesity-linked insulin resistance, were identified by immunostaining.     

Adrenalectomy and castration in the genetically obese (ob/ob) mouse

The present studies have tested the hypothesis that adrenalectomy could modify the phenotypic expression of genetic obesity by examining the effects of adrenalectomy on the function of the gonadal system in lean and ob/ob mice. Corticosterone concentrations were undetectable in the adrenalectomized animals. Adrenalectomy significantly slowed the weight gain of obese mice in comparison to sham-adrenalectomized controls. Gonadectomy had no independent effect on weight gain. The testes, prostate, and seminal vesicles in the ob/ob mice were significantly smaller than in the lean animals. Castration lowered the weights of the prostate and seminal vesicles in the lean mice to weights close to those observed in the castrated ob/ob mice. Castration significantly increased the concentrations of LH and FSH in both ob/ob and lean mice, but the absolute concentrations were higher in the lean mice in both conditions. Adrenalectomy per se had no effect on the concentration of LH, FSH, or testosterone or on the weights of the prostate or seminal vesicles. These data indicate that adrenalectomy has no effect on the physiologic control of the reproductive system in genetically obese mice, and are consistent with the hypothesis that the defect in the ob/ob mouse is a modulator of steroid action which over expresses glucocorticoid effects and under expresses gonadal steroid effects.     

Aspalathin improves hyperglycemia and glucose intolerance in obese diabetic ob/ob mice

Purpose

Although several researches have demonstrated that rooibos extract has hypoglycemic effect, the role of aspalathin, a main polyphenol in the extract, remains unclear. Our aims were to find specific mechanisms for anti-diabetic action of aspalathin employing a rat skeletal muscle-derived cell line (L6 myocytes) and a rat-derived pancreatic β-cell line (RIN-5F cells) and to investigate its effect in type 2 diabetic model ob/ob mice.

Methods

We investigated in vitro the effect of aspalathin on the glucose metabolism through the studies on molecular mechanisms of glucose uptake using cultured L6 myotubes. We also measured the antioxidative ability of aspalathin against reactive oxygen species (ROS) generated by artificial advanced glycation end product (AGE) in RIN-5F cells. In vivo, ob/ob mice were fed 0.1 % aspalathin-containing diet for 5 weeks, and the effect of aspalathin on fasting blood glucose level, glucose intolerance, and hepatic gene expression was studied.

Results

Aspalathin dose dependently increased glucose uptake by L6 myotubes and promoted AMP-activated protein kinase (AMPK) phosphorylation. Aspalathin enhanced GLUT4 translocation to plasma membrane in L6 myoblasts and myotubes. In RIN-5F cells, aspalathin suppressed AGE-induced rises in ROS. In vivo, aspalathin significantly suppressed the increase in fasting blood glucose levels and improved glucose intolerance. Furthermore, aspalathin decreased expression of hepatic genes related to gluconeogenesis and lipogenesis.

Conclusions

Hypoglycemic effect of aspalathin is related to increased GLUT4 translocation to plasma membrane via AMPK activation. In addition, aspalathin reduces the gene expression of hepatic enzymes related to glucose production and lipogenesis. These results strongly suggest that aspalathin has anti-diabetic potential.     

Apolipoprotein M

Apolipoprotein M (apoM) is a 26-kDa protein that is mainly associated with high-density lipoprotein (HDL) in human plasma, with a small proportion present in triglyceride-rich lipoproteins (TGRLP) and low-density lipoproteins (LDL). Human apoM gene is located in p21.31 on chromosome 6 (chromosome 17, in mouse). Human apoM cDNA (734 base pairs) encodes 188-amino acid residue-long protein. It belongs to lipocalin protein superfamily. Human tissue expression array study indicates that apoM is only expressed in liver and in kidney and small amounts are found in fetal liver and kidney. In situ apoM mRNA hybridization demonstrates that apoM is exclusively expressed in the hepatocytes and in the tubule epithelial cells in kidney. Expression of apoM could be regulated by platelet activating factor (PAF), transforming growth factors (TGF), insulin-like growth factor (IGF) and leptin in vivo and/or in vitro. It has been demonstrated that apoM expression is dramatically decreased in apoA-I deficient mouse. Hepatocyte nuclear factor-1α (HNF-1α) is an activator of apoM gene promoter. Deficiency of HNF-1α mouse shows lack of apoM expression. Mutations in HNF-1α (MODY3) have reduced serum apoM levels. Expression of apoM is significantly decreased in leptin deficient (ob/ob) mouse or leptin receptor deficient (db/db) mouse. ApoM concentration in plasma is positively correlated to leptin level in obese subjects. These may suggest that apoM is related to the initiation and progression of MODY3 and/or obesity.     

Iron metabolism in genetically obese (ob/ob) mice

Several reports in the clinical literature suggest that obese children may be at risk for developing iron deficiency. Here the absorption, retention, tissue distribution and tissue levels of iron were compared in lean (+/?) and obese (ob/ob) C57BL/6J mice to examine the impact of obesity on the iron status of this animal model. Obese mice absorbed and retained approximately twice as much 59Fe as lean mice after receiving a solution containing 1 mumol iron per os. This difference was independent of age, severity of obesity and mass of the gastrointestinal tract. Obese mice fed ad libitum had higher levels of 59Fe in blood and fat pads, but lower amounts of 59Fe in the skeletal-muscular system, than lean mice 6 d after subcutaneous injection of 1 mumol of the metal. At least 30% of carcass 59Fe was present in the liver of obese and lean mice 6 d after injection. Despite significantly lower concentrations of iron in liver and bone, blood hemoglobin and hematocrit were significantly higher in obese mice fed ad libitum than in lean mice at 10 wk of age. Plasma iron and transferrin were not affected by chronic obesity. Although several characteristics of iron metabolism differed in obese and lean mice, the results indicate that ob/ob mice were not iron deficient when fed a diet containing an adequate level of this micronutrient. The increased absorption of iron by obese mice probably represents an adaptive response that is required to supply additional micronutrient for the expanded blood volume in these animals.     

Hog millet (Panicum miliaceum L.)-supplemented diet ameliorates hyperlipidemia and hepatic lipid accumulation in C57BL/6J-ob/ob mice

Dietary intake of whole grains reduces the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. In an earlier study, we showed that Panicum miliaceum L. extract (PME) exhibited the highest anti-lipogenic activity in 3T3-L1 cells among extracts of nine different cereal grains tested. In this study, we hypothesized that PME in the diet would lead to weight loss and augmentation of hyperlipidemia by regulating fatty acid metabolism. PME was fed to ob/ob mice at 0%, 0.5%, or 1% (w/w) for 4 weeks. After the experimental period, body weight changes, blood serum and lipid profiles, hepatic fatty acid metabolism-related gene expression, and white adipose tissue (WAT) fatty acid composition were determined. We found that the 1% PME diet, but not the 0.5%, effectively decreased body weight, liver weight, and blood triglyceride and total cholesterol levels (P < 0.05) compared to obese ob/ob mice on a normal diet. Hepatic lipogenic-related gene (PPARα, L-FABP, FAS, and SCD1) expression decreased, whereas lipolysis-related gene (CPT1) expression increased in animals fed the 1% PME diet (P < 0.05). Long chain fatty acid content and the ratio of C18:1/C18:0 fatty acids decreased significantly in adipose tissue of animals fed the 1% PME diet (P < 0.05). Serum inflammatory mediators also decreased significantly in animals fed the 1% PME diet compared to those of the ob/ob control group (P < 0.05). These results suggest that PME is useful in the chemoprevention or treatment of obesity and obesity-related disorders.     

Suppressive Role of Lactoferrin in Overweight-Related Female Fertility Problems

The secretory glycoprotein lactoferrin (LF) is suggested to ameliorate overweight regardless of non-genetic or genetic mechanisms. Although maternal overweight represents a key predictor of offspring growth, the efficacy of LF on fertility problems in overweight and obese mothers remains unknown. To address this issue, we examined the effect of LF ingestion by analyzing overweight mice (Institute of Cancer Research (ICR) mice with high-fat diets; HF mice) and obese mice (leptin-deficient mice with type II diabetes; ob/ob mice). Plasma insulin, leptin, glucose, and cholesterol levels were measured, and thermal imaging and histological analysis were employed. The litter size of HF females was reduced due to miscarriage, which was reversed by LF ingestion. In addition, LF ingestion suppressed overweight prevalence in their offspring. The component analysis of the maternal blood demonstrated that glucose concentration in both HF females and their offspring was normalized by LF ingestion, which further standardized the concentration of insulin, but not leptin. LF ingestion was unable to reverse female infertility in ob/ob mice, although their obesity and uterine function were partially improved. Our results indicate that LF upregulates female fertility by reinforcing ovarian and uterine functions in females that are overweight due to caloric surplus.     

Sensitivity of ob/ob mice to leptin-induced adipose tissue apoptosis

OBJECTIVE: ob/ob mice have increased sensitivity to many of leptin's effects. The primary objective of this experiment was to determine whether ob/ob mice demonstrated increased sensitivity to leptin-induced adipose tissue apoptosis. RESEARCH METHODS AND PROCEDURES: Fifteen-week-old female ob/ob and Ob/? mice received 0 (saline), 2.5, or 10 microg/d leptin for 14 days through subcutaneous (sc) osmotic minipumps. Food intake (FI), body temperature, physical activity, and body weight were measured daily. Body composition and weights and adipose tissue apoptosis (percentage DNA fragmentation) of inguinal, parametrial, and retroperitoneal fat pads were determined at the end of the study. RESULTS: FI decreases were more pronounced in ob/ob. Leptin (10 microg/d) decreased total FI 71% in ob/ob and 34% in Ob/? (p < 0.05). Body weight was decreased by both doses of leptin in ob/ob (p < 0.01) but was unchanged in Ob/?. Leptin increased body temperature in ob/ob but not in Ob/?. Physical activity was increased 400% by 10 microg/d leptin in ob/ob (p < 0.01) but decreased 13% in Ob/? (p < 0.01). Body fat content of ob/ob was reduced by both leptin doses, whereas only 10 microg/d leptin decreased body fat in Ob/?. Fat pad weights were decreased similarly by leptin in both genotypes. However, apoptosis was increased by leptin in all three fat pads in ob/ob, whereas Ob/? showed significant increases only in retroperitoneal. DISCUSSION: ob/ob mice had greater overall sensitivity to leptin. Although ob/ob mice appeared to be more sensitive than Ob/? mice to leptin-induced adipose tissue apoptosis, there were differences among adipose depots in responsiveness to leptin-induced apoptosis.     

Dietary glucose and fat attenuate effects of adrenalectomy on energy balance in ob/ob mice.

The body energy balance response of ob/ob mice to adrenalectomy is diet dependent. Diets varying in source of carbohydrate (starch, glucose or fructose), fat to starch ratio, or fat to glucose ratio were fed to determine influences on energy balance in female adrenalectomized ob/ob mice. Adrenalectomy lowered food intake in all ob/ob mice to values comparable to those of lean mice independent of diet, but the percentage of dietary energy retained as body energy in adrenalectomized ob/ob mice was lowered to values comparable to those of lean mice only in mice fed starch or fructose, not in mice fed glucose or fat. Consumption of either high glucose or high fat diets blocked about one-half the overall effect of adrenalectomy on energy balance in ob/ob mice. Hyperinsulinemia was associated with the high efficiency of energy retention observed in adrenalectomized ob/ob mice fed glucose, but high efficiency of energy retention occurred in adrenalectomized ob/ob mice fed high fat diets without concomitant hyperinsulinemia. Adrenalectomy-induced changes in brain weight and protein content, body protein content and plasma thyroid hormone concentrations in ob/ob mice were independent of diet consumption. Combined adrenalectomy-ovariectomy diminished, but did not eliminate, the diet-dependent influences on energy balance in ob/ob mice. In conclusion, dietary glucose and dietary fat both attenuate effects of adrenalectomy on energy balance in ob/ob mice by promoting high efficiencies of energy retention.     

The relationship between protein turnover and energy balance in lean and genetically obese (ob/ob) mice.

1. Groups of lean and genetically obese (ob/ob) mice were adapted to varying energy intakes and the rates of total protein turnover in liver, gut and kidney were measured. 2. Lean mice gained less weight when fed above maintenance and lost less weight when fed below maintance than obese mice. 3. Hepatic protein turnover (mg/d) was sigmoidally related to digestible energy intake in lean mice but showed no significant changes with dietary intake in obese mice. 4. The changes in protein turnover resulted from changes in both the half-lives of protein synthesis and catabolism and in tissue protein content. 5. In the lean mice, protein turnover in kidney and gut was not significantly changed with increasing energy intake until the highest level was reached. 6. The findings suggest that protein turnover may be an important cycle for the regulation of energy balance in mice and that this cycle is impaired in the genetically obese (ob/ob) mice.     

Cordyceps militaris alleviates non-alcoholic fatty liver disease in ob/ob mice

BACKGROUND/OBJECTIVES

Non-alcoholic fatty liver disease (NAFLD) is becoming an important public health problem as metabolic syndrome and type 2 diabetes have become epidemic. In this study we investigated the protective effect of Cordyceps militaris (C. militaris) against NAFLD in an obese mouse model.

MATERIALS/METHODS

Four-week-old male ob/ob mice were fed an AIN-93G diet or a diet containing 1% C. militaris water extract for 10 weeks after 1 week of adaptation. Serum glucose, insulin, free fatty acid (FFA), alanine transaminase (ALT), and proinflammatory cytokines were measured. Hepatic levels of lipids, glutathione (GSH), and lipid peroxide were determined.

RESULTS

Consumption of C. militaris significantly decreased serum glucose, as well as homeostasis model assessment for insulin resistance (HOMA-IR), in ob/ob mice. In addition to lowering serum FFA levels, C. militaris also significantly decreased hepatic total lipids and triglyceride contents. Serum ALT activities and tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were reduced by C. militaris. Consumption of C. militaris increased hepatic GSH and reduced lipid peroxide levels.

CONCLUSIONS

These results indicate that C. militaris can exert protective effects against development of NAFLD, partly by reducing inflammatory cytokines and improving hepatic antioxidant status in ob/ob mice.     

The effects of dietary sucrose on opiate receptor binding in genetically obese (ob/ob) and lean mice

Weanling genetically obese (C57BL/6J-ob/ob) and lean (+/?) mice were given access to either a standard laboratory diet or the standard diet plus a 32% sucrose solution. At the end of a 4-week period, animals were sacrificed and opiate receptor binding determined. Both obese and lean mice given access to sucrose consumed approximately 30% more calories per day than animals given access to the standard diet alone. Obese animals consumed significantly more calories from the sucrose solution than the lean animals. Genetically obese animals weighed more than lean littermates throughout the course of the study. Differences in body weight due to sucrose supplementation in both genetically obese and lean mice were significant by day 10 and increased in magnitude until the termination of the study. Whereas there were no significant differences in specific opiate receptor binding (pmol 3H-naloxone bound/mg brain protein) between the genetically obese and lean animals, opiate receptor binding was significantly greater in genetically obese animals given access to sucrose than in obese animals which had access only to the standard diet. These data demonstrate that the sucrose-induced model of obesity functions in mice and that giving ob/ob mice access to sucrose in addition to a standard laboratory diet results in increased opiate receptor binding.     

瘦素改善2型糖尿病ob/ob小鼠糖和脂质代谢作用机制的探讨

目的 研究瘦素对2型糖尿病小鼠糖脂代谢及肝功能的改善作用,并从氧化应激、内质网应激和胰岛素抵抗等方面探讨其可能机制。方法 选用瘦素缺陷的ob/ob小鼠作为2型糖尿病模型,根据是否给予瘦素处理分为两组（1 mg/（kg·d）, 腹腔内注射）;选用野生型C57BL/6J小鼠为正常对照。测定血清甘油三酯（triglyceride,TG）和总胆固醇（total cholesterol,TC）反映脂质代谢状况,血清谷丙转氨酶（alanine transaminase,ALT）和谷草转氨酶（aspartate aminotransferase,AST）反映肝功能; 酶联免疫吸附实验测定胰岛素水平;Western blot检测氧化应激指标还原烟酰胺腺嘌呤二核苷酸磷酸氧化酶 4（nicotinamide adenine dinucleotide phosphate reduced oxidase 4,NOX4）和过氧化氢酶 （catalase,CAT）、内质网应激标志葡萄糖调节蛋白78（glucose-regulated protein 78,GRP78）水平,胰岛素调控糖脂代谢通路蛋白激酶B（protein kinase B,AKT）的激活。结果 与野生型小鼠比较,ob/ob小鼠血糖、TG和TC、ALT和AST活力以及胰岛素水平皆升高（均有P<0.05）;与ob/ob小鼠比较,经瘦素处理的ob/ob小鼠血糖、TG和TC、ALT和AST活力以及胰岛素水平皆下降（均有P<0.05）。与野生型小鼠比较,ob/ob小鼠肝脏NOX4和GRP78水平增高（均有P<0.05）,CAT和p-AKT水平降低（均有P<0.05）;与未经瘦素处理的ob/ob小鼠相比,经瘦素处理的ob/ob小鼠肝脏中NOX4、GRP78降低（均有P<0.05）,CAT和p-AKT升高（均有P<0.05）。结论 瘦素可改善2型糖尿病小鼠糖脂代谢和肝功能,作用机制可能与其减轻肝脏氧化应激和内质网应激以及恢复对胰岛素的敏感性有关。     

Dietary glucose increases plasma insulin and decreases brown adipose tissue thermogenic activity in adrenalectomized ob/ob mice

The purpose of this study was to determine whether consumption of a high glucose diet would increase plasma insulin concentrations and decrease brown adipose tissue metabolism in adrenalectomized ob/ob mice previously fed a high starch diet. Male sham-operated and adrenalectomized ob/ob and lean mice were fed a high starch diet for 12 d, then switched to a high glucose diet for the last 2 or 4 d of the 14- or 16-d feeding trials. Adrenalectomized ob/ob mice consumed 16% more energy and gained 50% more weight without an increase in oxygen consumption when switched from a high starch diet to a high glucose diet. Within 2 d after the switch to the high glucose diet, plasma insulin concentrations increased by 70% without any change in plasma glucose concentrations; brown adipose tissue metabolism, as assessed by GDP binding to brown adipose tissue mitochondria, was decreased by 26% 4 d after the diet switch. Sham-operated ob/ob and lean mice and adrenalectomized lean mice were minimally affected by the switch to the high glucose diet. The increase in plasma insulin concentrations in adrenalectomized ob/ob mice induced by the high glucose diet may contribute to the observed depression in brown adipose tissue metabolism.     

Expression of interleukin-6 is greater in preadipocytes than in adipocytes of 3T3-L1 cells and C57BL/6J and ob/ob mice

Inflammation plays a major role in the development of chronic diseases such as cardiovascular disease and Type 2 diabetes. Further, it was demonstrated that obese animals and humans have significantly higher levels of circulating proinflammatory cytokines, such as interleukin-6 (IL-6). The aim of this study was to determine whether adipose tissue could be a major source of circulating IL-6 in leptin-deficient obese (ob/ob) mice by comparing the expression of IL-6 in different tissues of ob/ob mice. Our secondary goal was to determine whether preadipocytes are the source of adipose tissue IL-6. The ob/ob mice had higher levels of plasma IL-6 (P < 0.05) and adipose tissue IL-6 mRNA (P < 0.05) compared with lean mice. Interestingly, IL-6 mRNA levels of liver and spleen were not different between ob/ob and lean mice, whereas adipose tissue IL-6 mRNA levels were higher in the ob/ob mice compared with lean mice (P < 0.05). In addition, we showed that IL-6 secretion from the adipose tissue stromal vascular fraction cells was higher than that from fully differentiated adipocytes (P < 0.001). We further demonstrated that 3T3-L1 preadipocytes had significantly higher levels of lipopolysaccharide (LPS)-stimulated IL-6 mRNA and IL-6 secretion than differentiated 3T3-L1 adipocytes. Taken together, these data suggest that adipose tissue and preadipocytes from the adipose tissue stromal vascular fraction may contribute significantly to the increased plasma IL-6 levels in ob/ob mice.     

Hypothalamic neuronal histamine regulates body weight through the modulation of diurnal feeding rhythm

Hypothalamic neuronal histamine and its H(1) receptor (H(1)-R), a leptin signaling pathway in the brain, regulate body weight and adiposity by affecting food intake and energy expenditure. Glucagon-like peptide-1 and/or corticotrophin-releasing hormone mediate leptin signaling to neuronal histamine. Leptin-induced suppression of food intake and upregulation of uncoupling protein-1 expression in brown adipose tissue were partially attenuated in histamine H(1)-R knockout (H(1)KO) mice. H(1)KO mice developed maturity-onset obesity. Hyperphagia and decreased energy expenditure assessed by the expression of uncoupling protein-1 mRNA were observed in older (48-wk-old) obese H(1)KO mice but not in younger (12-wk-old) non-obese H(1)KO mice. However, the diurnal feeding rhythm was impaired even in younger non-obese animals. Specifically, disruption of the feeding rhythm developed before the onset of obesity in H(1)KO mice. Correction of these abnormal feeding rhythms with scheduled feeding improved the obesity and associated metabolic disorders in the H(1)KO mice. These findings suggest that histamine H(1)-R is crucial for regulating the feeding rhythm and in mediating the effects of leptin. Early disruption of H(1)-R-mediated functions in H(1)KO mice may lead to hyperphagia and decreased energy expenditure, which may contribute to the development of obesity in these animals.     

The development of cold-induced thermogenesis and the structure of brown adipocyte mitochondria in genetically-obese (ob/ob) mice

The onset of cold-induced thermogenesis was studied in a strain of mice which produced among their offspring genetically-obese (ob/ob) individuals. A thermogenic response was present in a majority by day 5 after birth. The thermogenic response to cold was measured on days 5, 10 or 15 after birth, and the animals reared and the onset of obesity noted. The correlation between the subsequent development of obesity and a poor thermogenic response in early life was low. A poor thermogenic response at day 15 was associated with the presence in brown adipocytes of mitochondria with disordered internal structures. At day 42 both non-obese and obviously-obese mice showed a similar thermogenic response to moderate cold exposure. It would seem that in this strain of mice disordered internal mitochondrial structure in brown adipose tissue is associated with a poor thermogenic response to cold, but not invariably with the subsequent onset of obesity.     

Effect of high fat and high carbohydrate cafeteria diets on the development of the obese hyperglycemic (ob/ob) syndrome in mice

The role of dietary fat and carbohydrate in the development of obesity, impaired glucose homeostasis and hyperinsulinemia was examined in young obese-hyperglycemic (ob/ob) mice. Palatable and varied high fat (HF) and high carbohydrate (HC) cafeteria diets were fed to ob/ob mice from 3 to 10 weeks of age. Mice fed the HF cafeteria diet consumed 15% more energy and gained 50% more in body weight than mice fed the HC cafeteria diet. Metabolic efficiency, as indicated by the ratio of body weight gain to energy intake was increased by 29% in mice fed the HF cafeteria diet. The latter mice also showed an earlier onset of hyperglycemia and a greater impairment of glucose tolerance. The development of hyperinsulinemia and the loss of an insulin response to parenterally administered glucose were similar in the two groups of mice. The results indicate a greater preference for HF than HC cafeteria diets, a greater retention of energy consumed as fat, and a greater impairment of glucose homeostasis in young ob/ob mice fed the HF cafeteria diet.     

Tissue-Specific HSP70 Levels and Reproductive Physiological Responses in Fishes Inhabiting a Metal-Contaminated Creek

The 70-kDa stress protein family (HSP70) plays important roles in a variety of physiological processes, including protein chaperoning, protection against apoptosis, steroidogenesis, and general cellular stress responses in vertebrate organisms, and has also been proposed as a biochemical marker of environmental stress, such as toxicant exposure. The objectives of this study were to determine HSP70 protein expression in head kidney, liver, gill, and ovarian tissues and to examine reproductive physiological responses in female fishes exposed chronically to sublethal metal concentrations. Female black bullhead (Ameiurus melas) and bluegill sunfish (Lepomis macrochirus) were collected from Tar Creek, Oklahoma (flowing through the Tri-State mining district) and from a nearby reference creek (Lytle Creek) during spring (prespawning; 26.5 ± 0.95°C water temperature) and winter (ovarian recrudescence; 4.8 ± 0.80°C water temperature). Aqueous (dissolved and suspended) concentrations of Cd and Zn and liver concentrations of Cd and Zn in both fish species were significantly greater at Tar Creek compared to Lytle Creek. HSP70 expression was consistently elevated in the head kidney of both fish species collected at Tar Creek in comparison to fish collected from the reference creek. In contrast, no consistent differences were observed in HSP70 expression in liver, gill, or ovarian tissues between sites. Significant seasonal differences were observed in expression of HSP70 in gill tissue of both species, in ovarian and liver tissue of bluegill sunfish and in head kidney of black bullhead. Serum testosterone concentration was significantly reduced in sunfish collected from Tar Creek during winter. Gonadosomatic and hepatosomatic indices were significantly lower in black bullhead collected from Tar Creek during spring, and condition factors were lower in black bullhead collected from Tar Creek during both spring and winter. There was no significant difference in the extent of ovarian follicular cell apoptosis in either species collected during spring. In conclusion, we observed significant tissue specific differences and seasonal variation in expression of HSP70, as well as alterations in circulating testosterone levels in female fish chronically exposed to metals.Received: 12 May 2002/Accepted: 26 January 2003