LC Rats Research Articles

Deficit of β-endorphin neurons in the hypothalamus and high expression of MOR in mesolimbic structures are related to high alcohol consumption in outbred rats.

Clinical studies have postulated that β-endorphin deficiency generates excessive alcohol consumption, and it has been shown that the reduction of β-endorphin neurons increases alcohol intake in animal models. The β-endorphin produce their rewarding effect when they act mainly on the μ-opioid receptors (MOR) located in mesolimbic structures. Thus, it is possible that individual differences in these components of the endogenous opioid system are related to different levels of alcohol consumption. The present study thus examines the relation between two levels of alcohol consumption and intrinsic characteristics of the components of the opioid system in outbred Wistar rats that were not genetically selected. We analyzed the number of β-endorphin-positive neurons in the arcuate nucleus (ArN) and the expression of μ-opioid receptors (MOR) in regions of the reward system, such as the nucleus accumbens (NAc), amygdala (Amy), and ventral tegmental area (VTA) in outbred rats with low (LC) or high (HC) voluntary alcohol consumption. Findings showed that the HC rats had a lower number of β-endorphin-positive neurons in the hypothalamic ArN and a higher expression of MOR in the NAc and VTA, compared to the LC rats. No changes in the expression of MOR in the Amy were observed between the two groups. Results suggest that intrinsic variability in the number of β-endorphin neurons and in the expression of MOR in the LC and HC rats could explain their different patterns for alcohol intake.

Differential expression of μ-opioid receptors in the nucleus accumbens, amygdala and VTA depends on liking for alcohol, chronic alcohol intake and estradiol treatment

Affectations of the opioid system have been related to exacerbated alcohol consumption. The objectives of this work were to assess whether a deficit of β-endorphinergic neurons differentially affects alcohol intake in female rats with low (LC) and high alcohol consumption (HC), and to determine changes in the μ-opioid receptors (MOR) related to alcohol consumption and chronic exposure to alcohol in structures of the mesolimbic system. Female wild-type rats were selected according to their baseline alcohol intake levels and then exposed to chronic voluntary alcohol consumption after a single injection of either the vehicle or estradiol valerate (EV) to produce a β-endorphin neuronal deficit. Changes in alcohol consumption and MOR expression levels were assessed in the nucleus accumbens (NAc), amygdala (Amy) and ventral tegmental area (VTA) at 5 and 10 weeks after EV treatment. The LC rats increased alcohol intake from baseline to the initial weeks after EV treatment and this consumption remained stable throughout the studied period. In contrast, alcohol consumption increased steadily over time in the HC rats. The HC vehicle rats had a 38% higher MOR protein expression in the NAc than the LC vehicle rats. In addition, chronic alcohol consumption increased MOR expression in the Amy regardless of consumption level, whereas EV treatment produced a decrease in MOR expression in the VTA in all groups. These results suggest intrinsic differences in MOR expression related to alcohol consumption levels. Also, the EV treatment and chronic exposure to alcohol produced adaptive changes in MOR expression.

Acute-on-chronic liver disease enhances phenylephrine-induced endothelial nitric oxide release in rat mesenteric resistance arteries through enhanced PKA, PI3K/AKT and cGMP signalling pathways

Acute-on-chronic liver disease is a clinical syndrome characterized by decompensated liver fibrosis, portal hypertension and splanchnic hyperdynamic circulation. We aimed to determine whether the alpha-1 agonist phenylephrine (Phe) facilitates endothelial nitric oxide (NO) release by mesenteric resistance arteries (MRA) in rats subjected to an experimental microsurgical obstructive liver cholestasis model (LC). Sham-operated (SO) and LC rats were maintained for eight postoperative weeks. Phe-induced vasoconstriction (in the presence/absence of the NO synthase –NOS- inhibitor L-NAME) and vasodilator response to NO donor DEA-NO were analysed. Phe-induced NO release was determined in the presence/absence of either H89 (protein kinase –PK- A inhibitor) or LY 294002 (PI3K inhibitor). PKA and PKG activities, alpha-1 adrenoceptor, endothelial NOS (eNOS), PI3K, AKT and soluble guanylate cyclase (sGC) subunit expressions, as well as eNOS and AKT phosphorylation, were determined. The results show that LC blunted Phe-induced vasoconstriction, and enhanced DEA-NO-induced vasodilation. L-NAME increased the Phe-induced contraction largely in LC animals. The Phe-induced NO release was greater in MRA from LC animals. Both H89 and LY 294002 reduced NO release in LC. Alpha-1 adrenoceptor, eNOS, PI3K and AKT expressions were unchanged, but sGC subunit expression, eNOS and AKT phosphorylation and the activities of PKA and PKG were higher in MRA from LC animals. In summary, these mechanisms may help maintaining splanchnic vasodilation and hypotension observed in decompensated LC.

SAT-243 Investigation of Intervention Effect on Thrifty Phenotype of Low Birth Weight Rat

[OBJECTIVES] Low birthweight infants caused by fetal malnutrition acquire a thrifty phenotype, according to the development of health and disease (DOHaD) theory. When these children grow in a eutrophic environment, there is a mismatch of phenotype and environment, and it becomes a risk of various metabolic diseases. We aimed to develop an interventional method by creating a rat model exhibiting hyperinsulinemia with low birth weight due to low carbohydrate & calorie restriction during pregnancy and high fat diet (HFD) exposure after growth. Since metaborome analysis of rat blood revealed various metabolic changes, we investigated whether insulin resistance caused by mismatch is improved by methyl donor supplementation. [METHODS] Pups were obtained from mother rats fed with low carbohydrate & calorie restriction diet (LC) for the entire pregnancy period. For control, we used pups from mother rats who had fed standard diet (SC) ad libitum. We made a methyl donor diet in accordance with the previous report and fed ad libitum to mothers who were 3rd trimester of gestational period or breastfeeding. The male rats were divided into an HFD group and SC group at 4 weeks of age, and the HFD group rats were exposed to HFD for 18 weeks. Then, oral glucose tolerance test was carried out to examine blood glucose level, blood insulin concentration, and the expression analysis of mRNA in the liver after an overnight fast. [Results] HFD-exposed LC rats did not cause abrupt change in body weight or in body fat mass induce by HFD exposure even if supplementation to methyl donor was given to lactating maternal rat after giving birth. The blood insulin concentration after oral glucose loading which was high in the HFD-exposed LC rats, was reduced by methyl donor supplementation. In addition, the expression of elevated in FOXO1 and decreased in insulin receptor in the HFD-exposed liver was normalized with supplementation with methyl donor to the LC rats. [Conclusion] Supplementation of pregnant or postnatal methyl donor caused metabolic changes, normalizing abnormalities in the regulation of gene expression in the liver of low birth weight rats.

A lactate-based compound containing caffeine in addition to voluntary running exercise decreases subcutaneous fat mass and improves glucose metabolism in obese rats

Abstract We examined the effect of a lactate-based compound containing caffeine with voluntary running exercise on fat loss and metabolic improvement in diet-induced obese (DIO) rats. The rats treated with exercise training (Ex) and the compound (i.e., 1000 mg/kg body weight of sodium lactate and 36 mg/kg body weight of caffeine) with exercise training (ExLC) for 5 weeks showed decreased epididymal and scapular fat mass compared to the sedentary (S) rats, while LC rats displayed significantly decreased scapular fat mass compared to the Ex rats. The area under the curve for the LC rats during an oral glucose tolerance test was significantly lower compared with the S and Ex rats. Thus, administration of a lactate-based compound containing caffeine can effectively decrease fat mass and improve glucose tolerance even with low volume exercise training in DIO rats.

Juvenile Play Behavior in Neonatally Underfed and Sensory Stimulated Wistar Rats

Play development in juvenile rats depends on specific sensory signals integrated at cortical, limbic and brain stem levels to modulate motoric, metabolic, motivational and social responses. Neonatal undernourishment disrupts the morphological and functional organization of the brain for adaptive responses including play performance. These alterations may be restored by preweaning exposure to sensory-enriched environments. This study was designed to determine in four experimental groups, Control (LC), Underfed (LU), Control Ligated/Stimulated (LCS), and Underfed Ligated/Stimulated (LUS), whether changes in juvenile play of neonatally underfed male rats by the nipple-ligated procedure of F0 dams and/or the handling of F1 rats may restore the deficiencies in juvenile play performance. The pinning frequency values in LC, LCS and LUS groups consistently increased until reaching a significant peak between postnatal days (PDs) 25 and 50 and then gradually declining until PD 60, when the play in pairs was significantly higher compared with the play in groups that follows the same sequence but with lower values in the stimulated groups. The results may reflect poor maternal care and lower somatosensory stimulation; and the sensory massage of LU F1 pups compared with the LC, LUS, and LCS rats. Fewer dorsal body contacts occurred in LU and LUS rats when playing in pairs than in groups. Results suggest that although handling has salutary effects on neuronal play structures, the reduced levels of total pinning and dorsal contacts, mainly in the play of rat pairs in LCS vs. LUS groups, were not fully recovered.

Chronic stress sensitizes amphetamine-elicited 50-kHz calls in the rat: Dependence on positive affective phenotype and effects of long-term fluoxetine pretreatment

High level of positive affectivity acts as a protective factor against adverse effects of stress and decreases vulnerability to mood disorders and drug abuse. Fifty-kHz ultrasonic vocalizations (50-kHz USV) index the level of positive affect in the rat, whereas stable, trait-like inter-individual differences in terms of vocalization activity exist. Previously we have demonstrated that chronic stress can alter the effect of repeated amphetamine administration on 50-kHz vocalizations, and this effect is different in rats with high and low positive affectivity. In the present study it was tested whether the chronic stress effect on amphetamine-induced 50-kHz USV activity is altered by inhibition of serotonin reuptake. Male Wistar high (HC) and low (LC) 50-kHz vocalizing rats were subjected to 43-day chronic variable stress (CVS) regimen. On day 17 of the CVS, the four-week once a day fluoxetine (10 mg/kg) treatment was started. After the CVS and fluoxetine treatment, amphetamine (1 mg/kg) was daily administered for ten days and again nine days after withdrawal. Chronically stressed rats developed cross-sensitization of 50-kHz USV-s with repeated administration of amphetamine except the stressed LC rats that had not received fluoxetine. Amphetamine treatment decreased serotonin turnover in the fluoxetine-treated HC rats, but increased it in fluoxetine-treated LC rats. The effect of amphetamine on levels of amino acids in frontal cortex and hippocampus also depended on previous experience with chronic stress, repeated treatment with fluoxetine, and positive affectivity. Hence, this study provides further evidence the effects of chronic stress, psychostimulants, and a selective serotonin reuptake inhibitor are influenced by the inherent positive affectivity.

Mapping trait-like socio-affective phenotypes in rats through 50-kHz ultrasonic vocalizations.

Fifty-kilohertz ultrasonic vocalizations (USV) in rats are believed to express inter-individual differences in trait-like positive affective phenotypes. Emission of 50-kHz USV can be induced by amphetamine (AMPH) to model mania-like positive affect, raising the possibility that predispositions for high 50-kHz USV production confer susceptibility to mania-like states. Such 50-kHz USV presumably express the sender's motivation for social contact and elicit social approach behavior in receivers. We recently showed that AMPH-induced 50-kHz USV are paralleled by mania-like patterns of enhanced social approach behavior towards playback of 50-kHz USV. Here, we assessed whether these AMPH effects are dependent on trait-like inter-individual differences in 50-kHz USV production. To this aim, we subdivided juvenile rats into those emitting low (LC) and high (HC) rates of baseline 50-kHz USV and compared them across four AMPH dosage conditions: 0.0, 0.5, 1.0, and 2.5mg/kg. HC rats were considerably more susceptible to AMPH in inducing 50-kHz USV than LC rats, consistently across all examined doses. They further appeared to attribute more incentive salience to signals of rewarding social contact, as evidenced by enhanced social approach behavior towards 50-kHz USV playback, a response pattern also seen in LC rats after receiving AMPH treatment. HC but not LC rats emitted aversive 22-kHz USV following 50-kHz USV playback, indicating increased proneness to experience negative affective states if no actual social consequence followed the incentive signal. Inter-individual differences in 50-kHz USV map onto a unique trait-like socio-affective phenotype associated with enhanced emotional reactivity towards social and non-social reward, possibly conferring risk to mania-like states.

Early Treatment With Enalapril and Later Renal Injury in Programmed Obese Adult Rats.

Obesity-related kidney disease should be prevented or retarded. We aimed to investigate whether early treatment with enalapril ameliorates later renal injury induced by early postnatal overnutrition. Three or ten male pups per mother were assigned to either the Obese or Lean group during the first 21 days of life. These pups were treated with enalapril (Obese enalapril, OE; Lean enalapril, LE) or vehicle (Obese control, OC; Lean control, LC) for 15-28 days. Body weight, blood pressure (BP), and renal alterations were determined at 3 months. Enalapril decreased body weight only in the Lean group at 3 months (P < 0.05). Systemic BP levels were higher in the LE, OC, and OE groups than in the LC group at 3 months (P < 0.05). Fewer glomeruli per section area were found in the LE, OC, and OE groups than in the LC group and in the OE group than in the OC group (P < 0.05). The LE and OE groups had higher index scores of glomerulosclerosis and tubulointerstitial fibrosis than the controls (P < 0.05). LE pups showed increased intrarenal angiotensin II receptor type (AT)2 and matrix metalloproteinase (MMP)-9 and decreased renin and tissue inhibitor of MMP (TIMP)-1 expression than the LC rats (P < 0.05). OE pups showed increased intrarenal AT2 and decreased AT1 and TIMP-1 expression than the OC rats (P < 0.05). In conclusion, early treatment with enalapril can induce detrimental renal effects in later life and may not be renoprotective in programmed obese adult rats. J. Cell. Physiol. 232: 447-455, 2017. © 2016 Wiley Periodicals, Inc.

Plasma amino acid and metabolite signatures tracking diabetes progression in the UCD-T2DM rat model.

Elevations of plasma concentrations of branched-chain amino acids (BCAAs) are observed in human insulin resistance and type 2 diabetes mellitus (T2DM); however, there has been some controversy with respect to the passive or causative nature of the BCAA phenotype. Using untargeted metabolomics, plasma BCAA and other metabolites were assessed in lean control Sprague-Dawley rats (LC) and temporally during diabetes development in the UCD-T2DM rat model, i.e., prediabetic (PD) and 2 wk (D2W), 3 mo (D3M), and 6 mo (D6M) post-onset of diabetes. Plasma leucine, isoleucine, and valine concentrations were elevated only in D6M rats compared with D2W rats (by 28, 29, and 30%, respectively). This was in contrast to decreased plasma concentrations of several other amino acids in D3M and/or D6M relative to LC rats (Ala, Arg, Glu, Gln, Met, Ser, Thr, and Trp). BCAAs were positively correlated with fasting glucose and negatively correlated with plasma insulin, total body weight, total adipose tissue weight, and gastrocnemius muscle weight in the D3M and D6M groups. Multivariate analysis revealed that D3M and D6M UCD-T2DM rats had lower concentrations of amino acids, amino acid derivatives, 1,5-anhydroglucitol, and conduritol-β-opoxide and higher concentrations of uronic acids, pantothenic acids, aconitate, benzoic acid, lactate, and monopalmitin-2-glyceride relative to PD and D2W UCD-T2DM rats. The UCD-T2DM rat does not display elevated plasma BCAA concentrations until 6 mo post-onset of diabetes. With the acknowledgement that this is a rodent model of T2DM, the results indicate that elevated plasma BCAA concentrations are not necessary or sufficient to elicit an insulin resistance or T2DM onset.

Nutritional recovery with a soybean diet after weaning reduces lipogenesis but induces inflammation in the liver in adult rats exposed to protein restriction during intrauterine life and lactation.

We evaluated the effects of postweaning nutritional recovery with a soybean flour diet on de novo hepatic lipogenesis and inflammation in adult rats exposed to protein restriction during intrauterine life and lactation. Rats from mothers fed with protein (casein) in a percentage of 17% (control, C) or 6% (low, L) during pregnancy and lactation were fed with diet that contained 17% casein (CC and LC groups, resp.) or soybean (CS and LS groups, resp.) after weaning until 90 days of age. LS and CS rats had low body weight, normal basal serum triglyceride levels, increased ALT concentrations, and high HOMA-IR indices compared with LC and CC rats. The soybean diet reduced PPARγ as well as malic enzyme and citrate lyase contents and activities. The lipogenesis rate and liver fat content were lower in LS and CS rats relative to LC and CC rats. TNFα mRNA and protein levels were higher in LS and CS rats than in LC and CC rats. NF-κB mRNA levels were lower in the LC and LS groups compared with the CC and LC groups. Thus, the soybean diet prevented hepatic steatosis at least in part through reduced lipogenesis but resulted in TNFα-mediated inflammation.

Changes in mitochondria fusion protein-2 hepatic expression in conditions of liver cirrhosis and acute on chronic liver failure

To determine the differential protein and mRNA expressions of mitochondria fusion protein-2 (Mfn2) in hepatic tissues in conditions of cirrhosis and acute on chronic liver failure using rat model systems,and to determine the correlative effects on production of adenosine triphosphate (ATP) and reactive oxygen species (ROS). A liver cirrhotic rat model (LC rats) was established by intraperitoneal injection of carbon tetrachloride (CCl4,in vegetable oil),and these mice were subsequently used (10 weeks later) to establish the acute on chronic liver failure rat model (LF rats) by injecting lipopolysaccharide and D-amino-galactose.Control groups (normal controls,NC rats) were established for each model by intraperitoneal injection of vegetable oil only.Protein expression of Mfn2 in liver was quantified by western blotting with fluorescence densitometry and immunofluorescence staining,and mRNA expression was measured by real-time fluorescence quantitative PCR.ROS levels in liver were measured by fluorescence spectrophotometry,and ATP content was measured by bioluminescence assay.Significance of inter-group differences was assessed by one-way ANOVA,and correlations were determined using bivariate statistical modeling. Mfn2 protein expression was significantly lower in the liver tissues from modeled rats than that from the control rats (LC:0.051+/-0.004 and LF:0.037+/-0.007 vs.NC:0.254+/-0.008;F=444.98,P less than 0.05).The mRNA expression followed the same trend of lower expression (LC:21.21+/-0.93 and LF:24.35+/-0.85 vs.NC:19.09+/-0.69; F=66.941,P less than 0.05).The ATP content in liver tissues was also significantly lower in the modeled rats (LC:2.07+/-0.05 mol/L and LF:1.81+/-0.11 mol/L vs.NC:3.24+/-0.08 mol/L; F =574.21,P less than 0.05).Lower Mfn2 expression was correlated with lower ATP content (r =0.982) and higher ROS content (r =0.803). Reduced Mfn2 expression in liver tissue may cause a decrease in ATP synthesis and increase in ROS generation,thereby disrupting metabolism and increasing oxidative stress in the liver under conditions of cirrhosis and liver failure.

Pharmacokinetic changes of drugs in a rat model of liver cirrhosis induced by dimethylnitrosamine, alone and in combination with diabetes mellitus induced by streptozotocin.

Rats with liver cirrhosis induced by N-dimethylnitrosamine (LC) and rats with LC with diabetes mellitus induced by streptozotocin (LCD) have been developed as animal models for human liver cirrhosis and liver cirrhosis with diabetes mellitus, respectively. Changes in the pharmacokinetics of drugs (mainly non-renal clearance, CLNR) in LC and LCD rats reported in the literature compared with respective control rats were reviewed. This review mainly explains the changes in the CLNRs of drugs (which are mainly metabolized via hepatic microsomal cytochrome P450s, CYPs) in LC and LCD rats, in terms of the changes in in vitro hepatic intrinsic clearance (CLint; mainly due to the changes in CYPs in the disease state), free (unbound) fraction of a drug in the plasma (fp) and hepatic blood flow rate (QH) depending on the hepatic excretion ratio of the drug. Generally, changes in the CLNRs of drugs in LC and LCD rats could be well explained by the above-mentioned three factors. The mechanism of urinary excretion of drugs (such as glomerular filtration or renal active secretion or reabsorption) in LC and LCD rats is also discussed. The pharmacokinetics of the drugs reported in the LC and LCD rats were scarce in humans. Thus, the present rat data should be extrapolated carefully to humans.

Intrauterine protein restriction combined with early postnatal overfeeding was not associated with adult-onset obesity but produced glucose intolerance by pancreatic dysfunction

We investigated if whether intrauterine protein restriction in combination with overfeeding during lactation would cause adult-onset obesity and metabolic disorders. After birth, litters from dams fed with control (17% protein) and low protein (6% protein) diets were adjusted to a size of four (CO and LO groups, respectively) or eight (CC and LC groups, respectively) pups. All of the offspring were fed a diet containing 12% protein from the time of weaning until they were 90 d old. Compared to the CC and LC groups, the CO and LO groups had higher relative and absolute food intakes, oxygen consumption and carbon dioxide production; lower brown adipose tissue weight and lipid content and greater weight gain and absolute and relative white adipose tissue weight and absolute lipid content. Compared with the CO and CC rats, the LC and LO rats exhibited higher relative food intake, brown adipose tissue weight and lipid content, reduced oxygen consumption, carbon dioxide production and spontaneous activity, increased relative retroperitoneal adipose tissue weight and unaltered absolute white adipose tissue weight and lipid content. The fasting serum glucose was similar among the groups. The area under the glucose curve was higher in the LO and CO rats than in the LC and CC rats. The basal insulinemia and homeostasis model assessment of insulin resistance (HOMA-IR) were lower in the LO group than in the other groups. The total area under the insulin curve for the LO rats was similar to the CC rats, and both were lower than the CO and LC rats. Kitt was higher in the LO, LC and CO groups than in the CC group. Thus, intrauterine protein restriction followed by overfeeding during lactation did not induce obesity, but produced glucose intolerance by impairing pancreatic function in adulthood.

Postweaning low-calcium diet promotes later-life obesity induced by a high-fat diet

The aim of this study was to investigate the effects of a postweaning low-calcium diet on later obesity and explore the underlying mechanisms. Ninety-six male rats were weaned at 3 weeks of age, fed standard (STD: 0.50% calcium, n=48) and low-calcium (LC: 0.15% calcium, n=48) diets for 3 weeks, and then fed the standard diet for a 3-week washout period successively. Finally, the STD rats were divided into STD control and high-fat diet (HFD) groups, and the LC ones into LC control and LC+HFD (LCHF) groups. The STD and LC rats were fed the standard diet, while the HFD control and LCFD ones were fed a high-fat diet for 6 weeks to induce obesity. During the three feeding periods, adenosine-monophosphate-activated protein kinase (AMPK) and its responsive proteins phospho-acetyl-coA carboxylase, carnitine palmitoyltransferase 1 and uncoupling protein 3 were persistently down-regulated in the LC group (decreased by 18%, 24%, 18% and 20%, respectively) versus the STD group, and these effects were significantly more pronounced in the LCHFD group (decreased by 21%, 30%, 23% and 25%, respectively) than the HFD group by a later high-fat stimuli, causing more fat and body weight in adulthood. However, lipolysis enzymes, serum leptin, insulin and lipids were not significantly affected until the body weight and fat content changed at 15 weeks of age. The results suggest that the low-calcium diet after weaning promotes rat adult-onset obesity induced by high-fat diet, which might be achieved by programming expressions of genes involved in AMPK pathway.

Splanchnic sympathectomy prevents translocation and spreading of E coli but not S aureus in liver cirrhosis

IntroductionSpontaneous bacterial peritonitis (SBP) is mainly caused by bacterial translocation of enteric Gram-negative bacteria, predominantly Escherichia coli. The sympathetic nervous system (SNS) is activated in advanced cirrhosis, particularly in the...

Programming of rat adrenal medulla by neonatal hyperleptinemia: adrenal morphology, catecholamine secretion, and leptin signaling pathway

Leptin serum concentration in early life is an important factor for adequate future development of the offspring. Previously, we demonstrated that hyperleptinemia on lactation programmed for hyperleptinemia, central leptin resistance with lower expression of the long form of leptin receptor at hypothalamus, and higher medullary catecholamine levels with cardiovascular consequences at adulthood. The central objective of this study was to determine the direct effect of leptin on adrenal medullary function of adult rats that were leptin treated during lactation. Adrenal morphology was also accessed. Recombinant murine leptin was injected in the pups during the first 10 days of life (group L, leptin-programmed) or at adulthood during 6 days (group LC). The controls of both experiments received saline (groups C and CC). Both treatments resulted in hyperleptinemia at 150 days old (+78% and 2-fold increase, respectively; P < 0.05). Programmed animals showed hypertrophy of adrenal and higher adrenal catecholamine content at 150 days old (3-fold increase, P < 0.05), and no changes were observed in the LC group. However, LC rats had lower adrenal content of tyrosine hydroxylase (-17%, P < 0.05). Leptin-programmed rats had a lower response to leptin in vitro stimulation (-22%, P < 0.05) and lower expression of key proteins of the leptin signaling pathway, leptin receptor and janus tyrosine kinase 2 in the medullas (-61% and -29%, respectively, P < 0.05). However, they presented higher expression of phosphorylated signal transducer and activator of transcription 3 (+2-fold, P < 0.05). Leptin treatment at adulthood did not affect these parameters. The higher catecholamine synthesis and secretion in the leptin-programmed rats observed in our previous study does not seem to be a consequence of the direct effect of leptin on the medullas. We suggest that the hyperleptinemia of the programmed animals increases adrenal medullary function through sympathetic nervous system activation. In conclusion, high leptin levels on lactation program the activity of the sympathoadrenal system at adulthood that may contribute to the development of adult chronic diseases such as hypertension.

Systemic spreading of intraperitoneal Staphylococcus aureus is increased in experimental liver cirrhosis but unaltered by splanchnic sympathectomy

In liver cirrhosis, nosocomial spontaneous bacterial peritonitis (SBP) is associated with poor prognosis and most frequently caused by gram-positive bacteria. We previously reported that the well-known hyperactivity of the sympathetic nervous system (SNS) in advanced cirrhosis contributes to the translocation of E. coli from the peritoneum to mesenteric lymph nodes and extraintestinal sites. However, the role of splanchnic SNS-activity for spreading of gram-positive challanges in cirrhosis remains unclear. Methods: Staphylococcus aureus (S. aureus 106 CFU/ml) was given i.p. (3ml) and 6 hours later mesenteric lymphnodes (MLN), liver, spleen, lung and peripheral blood were harvested sterile from ascitic cirrhotic rats (LC) and healthy controls (Con) with and without splanchnic sympathectomy (±SE) being achieved by anti-dopamine-ß-hydroxylase-saporin treatment i.p. Bacterial tissue burden was determined by standard microbiological culture techniques. Results: After S. aureus challenge LC rats presented with marked increases in incidence of S. aureus translocation to MLN and blood as compared to control rats (Table 1). Intraperitoneal application of S.aureus induced endogenous translocation of gram-negative bacteria to MLN, liver, spleen and lung in LC but not in healthy rats (Fig. 1). SE had neither impact on incidence nor severity of translocation of Staph.aureus to MLN, liver, spleen, lung or blood in either control or LC rats. However, SE tended to reduce translocation of endogenous gram-negative bacteria from the gut to liver, spleen and lung (Fig.1). Conclusions: In cirrhosis, i.p. application of S. aureus is associated with enhanced systemic spreading and induction of endogenous gram-negative bacterial translocation as compared to controls. Splanchnic sympathetic hyperactivity in cirrhosis seems not to contribute to the translocation of Staph.aureus but enhances gram-negative translocation.

Soybean diet alters the insulin-signaling pathway in the liver of rats recovering from early-life malnutrition

ObjectiveWe investigated if alterations in the insulin-signaling pathway could contribute to reduced hepatic glycogen levels in adult rats subjected to a protein deficiency during intrauterine life and lactation and reared through to recovery on a soybean diet. MethodsRats from mothers fed with 17% or 6% protein (casein) during pregnancy and lactation were maintained with a 17% casein diet (offspring born to and suckled by mothers fed a control diet and subsequently fed the same diet after weaning [CC group] and offspring born to and suckled by mothers fed a control diet and subsequently fed a soybean flour diet with 17% protein after weaning [CS group]), a soybean diet (offspring of mothers fed a low-protein diet and a control diet after weaning [LC group] and offspring of mothers fed a low-protein diet and fed a soybean flour diet containing 17% protein after weaning [LS group]), or a 6% casein diet (offspring of mothers fed a low-protein diet and subsequently fed the same diet after weaning [LL group]) from weaning until 90 d of life. ResultsA soybean diet did not modify basal serum glucose and glucagon concentrations, but raised basal serum insulin and consequently increased the serum insulin/glucose ratio. Insulin receptor and insulin receptor substrate-1 levels were lower in rats fed a soybean diet compared with those maintained with a casein diet. In the LS group, the p85 levels were higher than in the LC group, whereas in CS rats its expression was lower than in CC rats. The expression of p110 was lower in the CS group compared with the CC group and similar in the LS and LC groups. Insulin receptor substrate-1 phosphorylation was similar in the LS, LC, and CS groups and lower compared with the CC group. The insulin receptor substrate-1–p85/phosphatidylinositol 3-kinase association was lower in LS than in LC rats and in CS than in CC rats. Akt phosphorylation was lower in the CS and LS groups than in the CC and LC groups. ConclusionAdult rats maintained with a soybean diet exhibited insulin resistance due, at least in part, to alterations in the early steps of the insulin signal transduction pathway.

Alterations in mechanical properties of mesenteric resistance arteries in experimental portal hypertension

Splanchnic vasodilation is the pathophysiological hallmark in the development of the hyperdynamic circulatory syndrome in liver cirrhosis and portal hypertension. This has been attributed so far mainly to a marked vascular hyporeactivity to endogenous vasoconstrictors. However, myogenic tone and vessel stiffness have not been addressed in mesenteric arteries in liver cirrhosis. CCl(4)(-)-induced ascitic cirrhotic (LC) and age-matched control rats, portal vein-ligated (PVL) rats, and sham-operated rats were investigated. Third-order mesenteric resistance arteries were studied under no-flow conditions using a pressure myograph measuring media thickness and lumen diameter in response to incremental increases in intramural pressure, from which wall mechanics were calculated. Electron microscopy was used for investigation of wall ultrastructure, especially the fenestrae in internal elastic lamina (IEL). In PVL animals, no significant change in passive vessel strain, stress, media-to-lumen ratio, or cross-sectional area was noted. In contrast, in LC rats, vessel strain was markedly elevated compared with healthy control rats, indicating a marked reduction in vessel stiffness. In addition, the strain-stress curve was shifted to the right, and the elastic modulus in dependency on vessel stress decreased, demonstrating predominantly structure-dependent factors to be involved. The media-to-lumen quotient was not significantly altered, but cross-sectional area was highly increased in LC rats, indicating hypertrophic outward remodeling. These findings were paralleled by enlarged fenestrae in the IEL but no change in thickness of IEL or proportion of extracellular matrix or vascular smooth muscle in LC rats. We concluded that, in long-standing severe portal hypertension such as ascitic LC but not in short-term conditions such as PVL, mesenteric resistance arteries exhibit vascular remodeling and markedly less resistant mechanical properties, leading to decreased vessel stiffness accompanied by structural changes in the IEL. This may well contribute to the maintenance and severity of splanchnic arterial vasodilation in LC.