Carbohydrate supplementation before operation retains intestinal barrier function and lowers bacterial translocation in a rat model of major abdominal surgery

BACKGROUND: Overnight fasting of rats augments the susceptibility of the small intestine to ischemia-reperfusion damage. Feeding before surgery may improve injuries to distant organs that were induced by ischemia-reperfusion. The present study tested the hypothesis that one of the food constituents, namely carbohydrates, may be responsible for the protective effect of preoperative feeding on postoperative organ dysfunction. METHODS: Male Wistar rats were fed ad libitum for 5 d and had either free access to water or free access to a carbohydrate drink and water. Then they were fasted for 16 h and access remained to either water or a carbohydrate drink and water. Following this, the arteria mesenterica superior was clamped for 60 min followed by 180 min of reperfusion. Subsequently, the intestinal permeability of stripped ileum was determined by measuring the mucosal to serosal flux in Ussing chambers. For assessment of bacterial content, organs were aseptically removed and assessed for bacterial content by culture under anaerobic conditions. RESULTS: Preoperative supplementation with carbohydrates resulted in a better maintenance of intestinal barrier function when compared with water supplemented animals. Moreover, carbohydrate supplementation resulted in a reduction in the ischemiareperfusion-induced increase in bacterial content of the liver, kidney, and mesenteric lymph nodes. CONCLUSIONS: Preoperative intake of carbohydrates by rats retains both the intestinal barrier function and prevents translocation of bacteria to distant organs.     

Preoperative supplementation with a carbohydrate mixture decreases organ dysfunction-associated risk factors

BACKGROUND & AIMS: Recently, both asymmetrical dimethylarginine and IL-6 have been suggested to be associated with the induction and severity of single and multiple organ dysfunction. The aims of the present study were to elucidate if these factors were increased in an ischemia reperfusion (IR) model and whether pre-operative carbohydrate supplementation can reduce the risk factors along with the IR injury. METHODS: One group of male Wistar rats was fasted for 16 h (water ad libitum) prior to clamping the superior mesenteric artery (IR fasted n=14). A second group had ad libitum access to a carbohydrate solution prior to clamping (IR fasted CHO group n=11). Sham-fasted animals, which only received laparotomy and no clamping, served as controls (n=4). RESULTS: Plasma urea and ALAT activity were both increased in the IR fasted animals when compared to the sham rats (P=0.007 and P<0.02, respectively). Furthermore, it was shown that IR fasted rats had increased ADMA and IL-6 concentration in plasma when compared to sham animals (P<0.02). Moreover, the GSH level in lung was significantly decreased in the IR fasted animals (P=0.014). IR CHO supplemented showed no significant increase of ALAT activity and decrease of lung GSH. Furthermore, significantly lower plasma urea, ADMA and IL-6 concentration was seen in the IR CHO supplemented group when compared to the IR fasted rats (P=0.028, P<0.01 and P<0.02, respectively). The liver glycogen concentration in IR fasted rats was 48% of that IR rats supplemented the carbohydrate mixture. CONCLUSION: The present rat intestinal ischemia reperfusion model not only induces organ injury indicated by the classical parameters such as plasma urea and ALAT activity, but also increased plasma IL-6 and ADMA and decreased lung GSH concentration in IR fasted rats. Pre-operative supplementation with the carbohydrate mixture significantly lowered the plasma urea, IL-6 and ADMA concentrations and maintained lung GSH concentration. This indicates that pre-operative carbohydrate supplementation reduces post-operative organ injury.     

Preoperative fasting induced protection against renal ischemia/reperfusion injury is independent of ghrelin in mice

One of the factors negatively influencing the outcome after kidney transplantation is ischemia-reperfusion (I/R) injury. Preoperative fasting is able to confer protection against I/R injury. We hypothesized that the protection imposed by preoperative fasting is mediated by increased levels of acylated ghrelin. Male C57BL/6 mice, 10 to 12 weeks old, were fasted for 1, 2, or 3 days, after which, acylated ghrelin levels were determined. Ad libitum fed mice were injected with acylated ghrelin or phosphate-buffered saline before renal I/R injury. Furthermore, mice were fasted for 3 days during which they were injected with a growth hormone secretagogue receptor antagonist, to block the effects of ghrelin, or a vehiculum. Bilateral renal I/R injury was induced by clamping the artery and vein of the left and right kidney simultaneously for 37 minutes. Kidney function was assessed by means of serum urea values determined at 24 and 48 hours after reperfusion. Fasting significantly increased acylated ghrelin serum levels. Ghrelin suppletion in ad libitum fed animals or ghrelin receptor blockade in fasted animals did not affect renal function after I/R injury. Our data suggest that the increased levels of acylated ghrelin induced by fasting do not mediate its protection against renal I/R injury.     

Effect of carbohydrate feeding on insulin action in skeletal muscle after surgical trauma in the rat

Metabolic stress after surgery is associated with peripheral insulin resistance. Recent studies have suggested that preoperative glucose can ameliorate postoperative decreases in insulin-stimulated glucose disposal. In the present experiments, we used a bowel-resection model of surgical trauma to test the hypothesis that elevations of serum insulin induced by preoperative oral glucose or ad libitum feeding affects postoperative insulin-stimulated glucose uptake in skeletal muscle. Insulin-stimulated glucose transport was measured in vitro in soleus muscles after surgical trauma in fasted rats given oral glucose or water before surgery. Insulin-stimulated glucose transport was also assessed in vitro in fasted or fed traumatized rats and non-traumatized control animals. In addition, stress hormones (glucagon, corticosterone, and adrenaline) were measured before and after surgical trauma in fasted rats and rats fed ad libitum. In vitro skeletal-muscle insulin sensitivity and responsiveness were reduced postoperatively in fasted animals that received oral glucose loads before bowel resections and in rats fed ad libitum or fasted before surgery versus non-traumatized rats (all P < 0.05). Stress-hormone concentrations after trauma did not differ between fed and fasted animals. In the current study, insulin sensitivity and responsiveness were reduced in isolated skeletal muscles after bowel resection, but neither preoperative glucose supplementation nor free intake of mixed nutrients ameliorated the development of postoperative insulin resistance.     

Plasma volume in fasted pregnant rats

The effect of a two day fast on maternal plasma volume and plasma protein concentration was explored in pregnant rats. Fasting between days 17 and 19 of gestation prevented the rise in plasma volume observed in ad libitum fed rats. Plasma protein concentration declined between day 17 and day 19 in both fasted and not fasted rats. Placental weight was not reduced by fasting during this stage of gestation. Fetal weight in the fasted group was 91% of the fetal weight of the ad libitum fed group. These results show that acute undernutrition, like longer term food or protein restriction, can prevent adequate plasma volume expansion and slow fetal growth.     

Suppression of food intake by adenosine and inosine.

The effect of adenosine and inosine on food consumption was tested by a 4 hr satiety assay in rats fasted overnight who then received injections subcutaneously, and also in ad libitum fed rats infused intravenously for 24 hr periods. It was found that adenosine, and to a lesser degree inosine, produced a significant suppression of food intake (86 and 69% reduction from control, respectively) during the 1st hr after subbcutaneous injection. This effect persisted for 4 hr, at the end of which both substances produced an average 37% reduction in food intake from control values. This effect of adenosine was also found in ad libitum-fed animals infused intravenously with adenosine for 24 hr (35% reduction from control). These studies report a previously undescribed effect of adenosine and add another substance to the list of agents shown to influence food intake regulation. Since adenosine is known to play a local regulatory role on adipose tissue metabolism and is generated and released from adipose tissue, the present findings may indicate an additional role for adenosine in providing a feedback regulatory signal between the adipose organ and the hypothalamic centers of food intake regulation.     

Translocation of protein kinase C isoforms in rat muscle in response to fasting and refeeding.

Weanling rats were offered food ad libitum, or fasted for 18 h, or fasted and refed for times ranging from 5 to 30 min. Five protein kinase C (PKC) isoforms (alpha, epsilon, zeta, theta and mu) were detected in the hindlimb muscles by Western immunoblotting. PKC forms epsilon and theta were abundant in plantaris, but not in soleus muscle, and no difference in localization was detected between fed rats and those fasted for 18 h. PKC forms alpha and mu were affected by fasting and refeeding. PKC-mu was found only in the cytosolic fraction of the plantaris muscle of the fasted animal, but in the fully-fed animals it was also associated with the membrane fraction. The pattern of localization observed in the fully-fed state was restored in the fasted rats by 20 min refeeding. In contrast, PKC-alpha was not detected in the cytosolic fraction of the plantaris in fasted animals but rapidly reappeared there on refeeding, being restored to 20% and 80% of the fed value within 5 and 30 min of refeeding respectively. The timing of these changes was correlated with the increase in serum insulin concentration, which was significantly elevated above the fasted value by 5 min and at subsequent times. These data suggest a possible role for PKC isoforms alpha and mu in the metabolic changes that occur in skeletal muscle on transition between the fasted and the fed state.     

The effect of age and dietary restriction on bone strength, calcium and phosphorus contents of male F344 rats

At 6 weeks of age, male Fischer F344 rats were fed a purified, casein based diet either ad libitum or in the amount of 60% of the diet consumed by the rats, fed ad libitum (restricted diet). Femur bone tissues were obtained from the rats at 4 and 13 months of age. The femurs of the animals fed ad libitum were significantly stronger than the femurs of the animals fed a restricted diet both at the age of 4 and 13 months [p0.01]. The body weight and bone weight of the animals fed ad libitum were also higher than that of the animals on the restricted diet. The bone ash weight was significantly lower for food restricted animals [p0.013] when compared to the animals fed ad libitum. The bone phosphorus content was significantly higher in the diet restricted older animals [p0.02] when compared to the diet restricted younger animals. Results indicate that food restriction will delay bone maturation.     

Effects of preoperative flavonoid supplementation on different organ functions in rats

BACKGROUND: Previously it has been reported that preoperative feeding preserves heart function in rats after intestinal ischemia-reperfusion. To further improve postoperative organ function, bioactive nutrition compounds were selected in vitro against the xanthine oxidase radical cascade, an enzyme suggested to play a key role in the induction of single- or multiple-organ dysfunction. METHODS: Flavonoids were selected in vitro for their capacity to (1) inhibit xanthine oxidase, (2) scavenge superoxide, and (3) scavenge peroxylradicals. The most bioactive flavonoids were added to the preoperative nutrition to study their effect on postintestinal ischemia-reperfusion organ function. RESULTS: A combination of flavonoids selected on basis of effective flavonoid xanthine oxidase inhibition and superoxide scavenging resulted in increased superoxide scavenging. In vivo, the selected flavonoid mixture significantly lowered postischemic intestinal apoptosis and intestinal oxidative stress indicated by malondialdehyde concentration when compared with ischemia-reperfusion fasted and sham-fasted animals. Moreover, this flavonoid mixture significantly lowered plasma creatinine and urea concentration, both indicating a better postoperative kidney function. Furthermore, oxidative stress measured as this flavonoid mixture when compared with control significantly lowered plasma malondialdehyde concentration in fed rats. CONCLUSIONS: Coadministration of bioactive flavonoid mixture to preoperative nutrition, in contrast to fasting, attenuates ischemia-reperfusion injury by preserving kidney function in the rat and decreasing apoptosis in the intestine.     

Collagen production in fasted and food-restricted rats: response to duration and severity of food deprivation

Malnutrition is associated with defects in connective tissue metabolism such as altered growth and wound healing. Because collagen is the major protein in most tissues, we determined the threshold for induction of altered collagen production by partial food restriction in rats. Groups of animals were fasted 2 or 4 d or were fed 20-100% of a predetermined food intake for 4 to 8 d. Collagen and noncollagen protein production in articular cartilage were determined using purified collagenase digestion of collagen labeled for 2 h in vitro with [3H]proline. Significant decreases in collagen (P less than 0.01) were seen in rats after 4 d of 40% (weight-losing rats) or after 8 days of 80% (weight-gaining rats) ad libitum intake. Collagen production decreased with both duration and degree of food deprivation; after 8 d of 20% intake, collagen was less than 10% that of controls fed ad libitum (P less than 0.001). In contrast, noncollagen protein production was significantly decreased only after 4 or 8 d of less than 40% intake (weight-losing rats). Maximum suppression of noncollagen protein was to approximately 65% of levels in controls fed ad libitum (P less than 0.01) and was not further reduced in fasted rats. Insulin-like growth factor-I levels were significantly decreased with duration and severity of diet in parallel with changes in collagen. The degree and sensitivity of altered collagen production to small changes in food intake suggest close regulation of this peptide and a potential role for decreased collagen synthesis in connective tissues during mild states of undernutrition.     

Acute fasting before conception affects metabolic and endocrine status without impacting follicle and oocyte development and embryo gene expression in the rabbit

Food deprivation affects female reproduction. The goal of the present study was to elucidate in the rabbit model the effects of acute energy restriction on ovarian function (follicle development, atresia rate and in vitro oocyte maturation) and embryonic development and gene expression of some candidate genes. Serum metabolic parameters (non-esterified fatty acids (NEFA), triglycerides, glucose, insulin and leptin concentrations) and endocrine markers (oestradiol-17β and progesterone concentrations) were also studied. A control group of nulliparous does fed ad libitum and a 72-h fasted group were used. At the end of the nutritional treatment, the ovaries of half of the animals were retrieved while the other animals were re-fed and artificially inseminated to recover embryos at 84 h after insemination, during the luteal phase. At the end of fasting, increased serum NEFA and decreased leptin concentrations were observed in the fasted group, but no differences appeared in serum steroid concentrations, follicle population and atresia rate or nuclear and cytoplasmic oocyte maturation. In the luteal phase, insulin concentrations increased notably in the fasted group. The number of recovered embryos per female and the speed of embryo development were reduced in the food-deprived group. Acute fasting altered both metabolic and endocrine markers and embryo development, but follicle and oocyte development and embryo gene expression were not affected.     

The effect of long-term fasting on the branched chain acylcarnitines and branched chain carnitine acyltransferases.

The effect of fasting for 8 days on the levels of carnitine acyltransferases in heart, liver, liver mitochondria, skeletal muscle, skeletal muscle mitochondria, kidney, and testes in young adult male rats was determined. The specific activities of acetyl-, octanyl-, isobutyryl-, and isovaleryl-carnitine acyltransferase in mitochondria isolated from the livers of fasted animals were significantly higher than the levels of the transferases isolated from livers of fed animals. Similar results were obtained with the 500 x g supernatant fluids from liver. In contrast, the specific activities of carnitine acyltransferases of 500 x g supernatant fractions isolated from heart, skeletal muscle, kidney, and testes were the same for fed as fasted animals. The total carnitine content of liver, muscle, heart, and kidney was less in animals fasted for 8 days than in fed animals, but the amount/g of organ was higher in the animals fasted for 8 days. The amount of specific short-chain acylcarnitines in liver, muscle, and heart was determined for both fed and fasted animals. The amount of isobutyrylcarnitine and isovalerylcarnitine increased significantly in muscle from fasted animals. These data are consistent with the previous suggestion that carnitine may have a role in the metabolism of the branched-chain amino acids.     

Effects of medroxyprogesterone acetate and caloric restriction on the distribution of serum and liver lipids and proteins in female rats

The effects of a large dose (2 mg/100 g body weight) of medroxyprogesterone acetate (MPA), restricted feeding (50% of normal intake) and their interaction were investigated on the serum and liver lipid and protein distribution in female rats. MPA increased serum cholesterol and triglyceride levels in rats on ad libitum food intake without having any effect in the animals on restricted diet. There was no statistical difference in the total serum protein or albumin levels among the groups. The serum levels of alpha and gamma globulin were significantly lower in the underfed rats. MPA increased the concentration of gamma globulin in well nourished rats but interacted with dietary restriction to further reduce the level of this protein in the undernourished animals. There was no difference in the levels of hepatic phospholipids or free cholesterol among groups, However, cholesterol ester and triglyceride levels in liver were significantly elevated in the MPA-treated group fed ad libitum. A similar increase was not found in underfed rats. This study indicates that MPA exerts significant effects on the distribution of serum and hepatic lipids and serum globulin levels and that the effects are dependent upon the nutritional status of the subject.     

Effect of food restriction and intense physical training on estrous cyclicity and plasma leptin concentrations in rats

Intense physical training and dietary energy restriction have been associated with consequences such as nutritional amenorrhea. We investigated the effects of intense physical training, food restriction or the combination of both strategies on estrous cyclicity in female rats, and the relationship between leptin ad these effects. Twenty-seven female Wistar rats were distributed into four groups: SF: sedentary, fed ad libitum; SR: sedentary subjected to 50% food restriction (based on the food intake of their fed counterparts); TF: trained (physical training on a motor treadmill with a gradual increase in speed and time), fed ad libitum; TR: trained with 50% food restriction. We analysed estrous cyclicity, plasma leptin and estradiol as well as chemical composition of the carcass, body weight variation, and weight of ovaries and perirenal adipose tissue. Data demonstrate that physical training alone was not responsible for significant modifications in either carcass chemical composition or reproductive function. Food restriction reduced leptin levels in all animals and interrupted the estrous cyclicity in some animals, but only the combination of food restriction and physical training was capable of interrupting the estrous cyclicity in all animals. Leptin was not directly related to estrous cyclicity. From our findings, it may be concluded that there is an additive or synergistic effect of energy intake restriction and energy expenditure by intense physical training on estrous cyclicity. Leptin appears to be one among others factors related to estrous cycle, but it probably acts indirectly.     

Soluble dietary fiber and cholesterol influence in vivo hepatic and intestinal cholesterol biosynthesis in rats.

Ninety-six male Sprague-Dawley rats were randomly assigned to eight dietary treatments. Rats were fed, with ad libitum access, diets containing 10% dietary fiber as cellulose (control), pectin, psyllium or oat bran with or without 0.3% added cholesterol for 3 wk. Among cholesterol-fed rats, liver total cholesterol was significantly lower in rats fed diets supplemented with either pectin or psyllium compared with those fed cellulose. In contrast, rats fed oat bran with cholesterol had significantly higher liver cholesterol concentrations compared with cellulose-fed animals. Liver total lipid concentrations were significantly lower in groups fed pectin and psyllium with or without dietary cholesterol compared with cellulose-fed controls. Pectin feeding with or without dietary cholesterol significantly lowered plasma total cholesterol compared with cellulose feeding. Oat bran had no effect on plasma total cholesterol compared with control diets. Hepatic sterol synthesis was significantly greater for animals fed soluble dietary fibers compared with those fed cellulose, but the effect on intestinal sterol synthesis was less pronounced.     

Visceral organ size and hepatocyte metabolic activity in fed and fasted rats

Changes in visceral organ mass and hepatocyte metabolic activity in response to nutrient deprivation were studied in male Sprague-Dawley rats. Forty-two rats (320 g) were given ad libitum access to feed or fasted for 72 h after which time visceral organ mass and metabolic activity were measured. Liver metabolic activity was measured in vitro by [14C]valine incorporation into acid-precipitable protein and oxygen consumption in isolated hepatocytes. Fasted rats had lower weights of liver and intestines and similar kidney and stomach weights relative to body weight than fed rats. Compared with fed controls, fasted rats had lower RNA and higher DNA concentrations in liver and intestines with decreased RNA and protein mass in all visceral organs. Nutrient deprivation generally resulted in reduced ratios of RNA/DNA and protein/DNA in visceral tissues. Nutrient deprivation had no effect on in vitro oxygen consumption or [14C]valine incorporation expressed per g tissue. However, in vitro oxygen consumption and valine incorporation expressed per unit DNA and estimated total liver oxygen consumption were reduced in fasted rats. These data suggest that nutrient deprivation reduced visceral organ mass of RNA and protein primarily through an apparent reduction in cell size. Nutrient deprivation resulted in decreased liver oxygen consumption because of decreased organ size rather than by tissue metabolic activity.     

Effect of short fasts on the development of tibial dyschondroplasia in chickens

The length of time each day that feed is available to young broiler-type chickens significantly influenced the development of tibial dyschondroplasia. Birds that were fasted for 8 or 10 h per day had reduced incidences of tibial dyschondroplasia as compared to birds fed ad libitum (5-11% vs. 59-68%). When birds were fasted for only 2 or 4 h each day there was no significant effect on the development of tibial dyschondroplasia. Birds fasted for 8 h every other day or every 4th d had lower incidences of tibial dyschondroplasia compared to ad libitum fed controls. Fasting for 8 h on the 4th d or the 4th and 8th d of a 20-d experiment had no effect on the incidence of tibial dyschondroplasia as compared to controls. Except for the daily fasting there was little effect of fasting on 20-d body weight or gain per feed intake. Fasting increased the percent bone ash in the tibia in all experiments. Fasting had no effect on total calcium, ultrafilterable calcium, dialyzable phosphorus or growth hormone levels in plasma taken from the birds the 18th and 19th day. The hour of the day that the birds were bled had a significant influence on the plasma total calcium, dialyzable phosphorus and growth hormone levels.     

Intravenous feeding of the rat with short chain fatty acid esters. II. Monoacetoacetin.

Acetoacetic acid is oxidized independent of carnitine transport into the cell mitochondria and its monoglyceride is a water soluble compound. The latter was examined as an intravenous nutrient. The monoglyceride of acetoacetic acid was prepared from the acid anhydride, diketene, and glycerol and was found to be totally miscible with water. The nutritional properties of monoacetoacetin were investigated by continuous intravenous infusion of 25 or 50 g/kg body weight per day into ad libitum fed rats. The response of these animals was compared to normal and food restricted rats. All experimental animals survived the 7 day study period in good health and were free of detectable physiological and behavioral abnormalities. The test rats demonstrated weight gain dependent on the infusion rate while on an inadequate spontaneous oral food intake diet. Ketonuria, hyperketonemia and monoglyceride excretion in the urine accompanied the infusion of monoacetoacetin and were dependent on the infusion load. At 50 g/kg per day total plasma ketones were found to be comparable with the infusion load. At 50 g/kg per day total plasma ketones were found to be comparable with the maximum level obtained in fasted rats, but the acetoacetate concentration was relatively higher than in fasting. This was consistant with ketosis derived from the hydrolysis of the monoglyceride. It was concluded from these results that monoacetoacetin might be an asset for intravenous nutrition.     

A critical appraisal of the usefulness of perioperative nutritional support.

Preoperative malnutrition is often associated with poor postoperative outcome, yet there is no consensus about whether perioperative nutritional support reduces postoperative complications to the level occurring in well-nourished patients undergoing similar procedures. This is partly because reports evaluating effect of perioperative nutritional support on postoperative outcome vary widely in number of patients studied, primary diagnosis, and duration and quality of perioperative nutritional support. These concerns warrant caution in interpreting reported results, even of randomized studies. However, analysis of published reports suggests that when total parenteral nutrition (TPN) is given to malnourished patients in adequate amounts for greater than or equal to 7-15 d preoperatively, significant improvements in both nutritional status and postoperative clinical outcome are likely to occur. Preoperative total enteral nutrition (TEN) is as effective as TPN in improving postoperative clinical outcome. Postoperative TPN, TEN, and ad libitum oral nutrition are equally effective in reducing postoperative complications. Potential candidates for surgery for whom prompt initiation of preoperative TPN or TEN may reduce operative morbidity and mortality irrespective of nutritional status can be identified on admission.