Effects of dietary fatty acids on the early stages of neoplastic induction in the rat pancreas. Changes in fatty acid composition and development of atypical acinar cell nodules.

Diets enriched with fat, especially unsaturated fat, promote experimental pancreatic carcinogenesis, but little is known of the effects of individual fatty acids. The effect of stearic and oleic acid on pancreatic fatty acids and atypical acinar cell nodules (preneoplastic lesions) was studied in 14-day-old weanling male Leeds strain rats (n = 60) given the carcinogen azaserine. Rats were allocated to one of six groups: untreated controls (n = 10), 20% stearic acid diet (n = 10), 20% oleic acid diet (n = 10), carcinogen alone (n = 10), carcinogen plus 20% stearic acid diet (n = 10) or carcinogen plus 20% oleic acid diet (n = 10). Azaserine was administered by intraperitoneal injection in a dose of 30 mg/kg at 2, 3 and 4 weeks of age. When total lipid extracts of pancreas were examined, there was an increase in stearic acid in the stearic acid fed group and an increase in oleic acid in the oleic acid fed group, irrespective of carcinogen treatment. The relative content of all other pancreatic fatty acids was suppressed by feeding oleic acid. At 26 weeks, the number and volumetric indices of pancreatic atypical acinar cell nodules was increased only in rats given azaserine and oleic acid. The enhancing effect of oleic acid on pancreatic carcinogenesis may be associated with pancreatic fatty acid changes.     

Potentiation of ethanol-induced pancreatic injury by dietary fat. Induction of chronic pancreatitis by alcohol in rats.

Effects of sustained ethanol intoxication and dietary fat content on pancreatic morphology were investigated in the rat model implanted with gastrostomy catheters, which permitted continuous intragastric infusion of ethanol plus liquid diet containing one of three levels of corn oil: 5% (low-fat), 25% (high-fat), and 35% (extra-high-fat) of total calories. After various durations of infusion ranging from 30 to 160 days, the pancreatic histology was examined. Mean blood alcohol levels achieved in the low, high, and extra-high fat diet groups were similarly high: 210 +/- 120, 224 +/- 122, and 289 +/- 110 mg/dl. The average weight gain of these ethanol-fed groups during the first 8 weeks of experiments was 15.4 +/- 1.9, 19.6 +/- 8.0, and 14.9 +/- 5.2 g/wk, respectively, and was not statistically different from that of pair-fed controls infused with isocaloric amount of dextrose and respective diet, nor from that of age-matched animals given the regular chow. None of control animals showed abnormal pancreatic morphologic features except occasional mild steatosis in those fed the extra-high-fat diet. With the low dietary intake of unsaturated fat, chronic ethanol intoxication produced only mild pancreatic pathology such as steatosis and interstitial edema. Administration of ethanol and the high-fat and extra-high-fat diets caused hypogranulation and apoptosis of acinar cells. Focal lesions of chronic pancreatitis were also observed in 20% or 30% of ethanol-fed animals given the high-fat or extra-high-fat diet. These lesions were characterized by fat necrosis, mononuclear cell infiltration, fibrosis, acinar atrophy, ductal dilatation, and intraductal mucious or proteinacious plugs. The incidence of focal acute pancreatitis was less (7-20%) but appeared increased with higher dietary fat content. Induction of either acute or chronic pancreatitis was not correlated with plasma levels of triglycerides or cholesterol. These results demonstrate potentiation by dietary unsaturated fat of ethanol-induced pancreatic injury. This model possesses many features analogous to those seen in alcoholic pancreatic injury in man. The hyperlipidemia does not appear to be an important pathogenetic factor for ethanol-induced pancreatitis produced in this model.     

Effect of amygdaloid lesions on dietary intake of disproportionate amounts of amino acids

Male rats with bilateral electrolytic lesions in the anterior, medial or posterior aspects of the ventral amygdala and groups of intact rats were fed, in turn, basal, imbalanced or deficient amino acid diets involving threonine or isoleucine as the limiting amino acid, and then a low protein (6% casein) followed by a high protein (75% casein) diet. No change in food intake was observed in animals fed the threonine basal diet postoperatively. When the threonine or isoleucine imbalanced diet was substituted for the respective basal diet, animals with lesions in certain areas of the medial amygdala showed little or no depression in food intake of the imbalanced diets, while all other rats with amygdala lesions reduced their food intake markedly, as did intact controls, when fed such diets. All animals, however, curtailed their food intake of the deficient or high protein diets. The lack of responsiveness of the animals with medial amygdaloid lesions to the imbalanced diets suggests that these areas may be involved in a system regulating food intake of animals fed diets containing imbalanced amino acid mixtures.     

Potentiation of azaserine by cholestyramine in the rat

In the rat, when pancreatic growth is stimulated there is an increased incidence of spontaneous pancreatic neoplasms and marked potentiation of the pancreatic carcinogen azaserine. Since previous studies showed that cholestyramine caused pancreatic growth in this species we have now studied the effect of azaserine in rats fed soya flour diets containing cholestyramine. Two groups, each of eight rats, were fed either heated soya flour (HSF) or raw soya flour (RSF). Two further groups, each of 12 rats, received the same diets containing 2% cholestyramine (HSF + C, RSF + C). In each group, four rats received azaserine (30 mg/kg i.p.) and the remainder saline, weekly, for the first 5 weeks. Animals were killed after 24 weeks and pancreatic growth and the number and size of pancreatic neoplastic nodules was measured. RSF caused a significant increase in pancreatic weight, protein, RNA and DNA, compared with HSF and cholestyramine caused a further significant increase in pancreatic weight, protein and RNA but not DNA. Azaserine did not affect pancreatic growth. In azaserine-injected rats significantly more nodules were seen and the nodules were larger and the tumour burden greater in rats fed HSF + C than in rats fed HSF alone. However, the nodule count and other nodule parameters were not significantly different in RSF and RSF + C fed rats. It is concluded that 2% cholestyramine enhances pancreatic growth when added to soya flour diets and in rats fed HSF it potentiates the action of azaserine on the pancreas. It does not increase the potentiation of azaserine seen with RSF up to 24 weeks.     

Effects of hippocampal lesions on adaptive intake of diets with disproportionate amounts of amino acids

Bilateral double electrolytic overlapping lesions were placed in dorsal-lateral hippocampus of male 230 g rats, and their food intake responses to the ingestion of diets containing disproportionate amounts of amino acids were examined. Rats with such lesions and intact control rats maintained their normal intakes of the 6% casein basal diet or a threonine basal amino acid diet postoperatively. However, they exhibited marked initial food intake depression, similar to that of intact rats, when fed the threonine imbalanced amino acid diet. Also, animals with lesions in certain areas of the dorsal-lateral hippocampus showed facilitated adaptation to the amino acid imbalanced diet. Similar severe reduction in food intake with relative lack of adaptation were observed in both the intact controls and rats with hippocampal lesions when fed amino acid diets completely devoid of threonine. Initial food intake of rats with hippocampal lesions was inhibited drastically as was the case with the intact controls when fed a 75% casein high protein diet. All rats, either intact or lesioned, showed similar slow adaptation patterns with the prolonged ingestion of the high protein diet. The initial food intake responses and facilitated adaptation of the animals bearing lesions in certain areas of the hippocampus suggest that such areas are not crucially involved in the inhibition of food intake of rats fed disproportionate amounts of dietary amino acids. Rather, such areas of lesions in the hippocampus may play a role in a system governing the behavioral adaptation of the intake of amino acid imbalanced diets but not of diets containing amino acids in general excess. This would also indicate that different mechanisms control the intake of amino acid imbalanced diets and diets containing amino acids in excess.     

Protein selection, food intake, and body composition in response to the amount of dietary protein

Though not universally observed, moderately low-protein diets have been found to increase caloric intake and body fat. It appears that animals overeat in calories in order to obtain more dietary protein. For animals to control protein intake, they must be able to distinguish between two isocaloric diets containing different percentages of protein and make the appropriate dietary selection on the basis of their previous history of protein intake. Experiment 1 examined the 24-h diet selection (5 vs. 35% casein) of Sprague-Dawley rats that had been previously fed diets containing various percentages of dietary protein (5, 10, 20, 35, or 60% casein). Animals fed 5, 10, or 20% dietary protein showed a preference for the higher protein selection diet. In contrast, no significant diet preference was found in animals pre-fed the two higher levels of dietary protein (35 or 60% casein). In this study, daily food intake and body fat of rats fed the low-protein diets (5 and 10% casein) were similar to rats fed the 20% casein diet. Experiment 2 examined the effects of the level of methionine supplementation on rats fed 10% casein. In this study, food intake and body fat were increased by approximately 20% in rats fed 10% casein diets, regardless of the level of methionine supplementation (0.3 vs. 0.15%). Together, the results suggest that the presence of low-protein-induced hyperphagia helps maintain body protein levels in the face of moderately low dietary protein and promotes an increase in the amount of body fat and energy.     

Effects of carbon tetrachloride and azathioprine on diethylnitrosamine and N-2-fluorenylacetamide-induced hyperplastic liver nodule and hepatocellular carcinoma

Effects of carbon tetrachloride (CCl4) and azathioprine (AZP) on the evolution of hyperplastic liver nodules and foci and hepatocellular carcinoma (HCC) were tested in short- and long-term in vivo experiments. In diethylnitrosamine (DEN)-treated rats, which were fed a N-2-fluorenylacetamide (FAA)-containing diet and additionally treated with repeated CCl4 injections, gamma-glutamyl transpeptidase (gamma-GTP)-positive hyperplastic nodules were markedly developed in the 8th week of the experiment. However, their number and area in liver sections were remarkably small in DEN-treated rats fed a diet containing both FAA and AZP. Increased area of gamma-GTP-positive foci was also observed in the 12th week in DEN-injected rats fed a choline-devoid died alone or treated with repeated doses of CCl4 alone. Hepatocellular carcinoma in DEN-injected rats treated with both FAA and CCl4 was first detected in the 21st week, and the incidence up to the 36th week was very high. However, no hepatocellular carcinoma developed in DEN-injected rats treated with both FAA and AZP. The increased activity of liver aniline hydroxylase observed 12 h after the administration of FAA, AZP or DEN alone was not observed when AZP was administered simultaneously with FAA to DEN-injected rats. The mechanisms of the effects of CCl4 and AZP on hepatocarcinogenesis are discussed with special reference to drug interaction.     

Smooth endoplasmic reticulum proliferation and increased cell multiplication in cultured hepatocytes of the newborn rat in the presence of phenobarbital

Twenty-eight-day old male Sprague-Dawley rats were fed diets containing 2, 5, 10, 15, 25, or 50% protein and varying levels of fat and energy for 8 weeks and were then killed. The livers of rats fed the 2% lactalbumin protein diet ad libitum but not the others showed visible fat. This difference was confirmed histologically. Chemical analyses of the livers indicated that the rats fed the 2% protein diets ad libitum had experienced a substantial reduction in liver cell size but only a very slight reduction in the lipid content of each cell. As a consequence, their livers showed a net increase in the amount of lipid per g of tissue and this gave them a fatty appearance. The lipids formed large fat globules suspended in a greatly reduced amount of protoplasm; there were no fat globules outside the cells. Therefore, the fatty liver of protein deficiency was a physical phenomenon rather than a metabolic defect. Rats fed the 2% protein diets in restricted amounts also had shrunken cells but they did not manifest fatty liver because there was a relatively greater reduction in liver cell lipid than in liver cell size. There was a reduced amount of protein in the livers of rats fed 2 and 5% protein diets which can be explained by a reduction in liver RNA and in Protein/RNA ratio. Protein/RNA ratio was enhanced by starvation. Liver protein content showed a biphasic response to variation in dietary fat level with its lowest value in rats fed diets containing 11.9% fat.     

Multiple atypical acinar cell nodules of the pancreas

Although acinar cell nodules of the pancreas have been described in rats given carcinogenic chemicals, similar nodules have not been reported in humans. This report describes two cases of nodular acinar cell lesions in the pancreas in patients who died of insulin secreting islet cell adenoma and of bronchogenic carcinoma. The lesions consisted of multiple nodules that were well demarcated from the surrounding acinar tissue and were composed of zymogen granule containing acinar cells with a pale to pink cytoplasm. The significance of these atypical acinar cell nodules in regard to their being possible precursor lesions of acinar cell carcinoma of the pancreas is discussed.     

Atypical acinar cell nodules of the human pancreas

The earliest morphological evidence of altered growth potential of pancreatic acinar cells of the rats treated with carcinogen, such as azaserine, is the development of nodules of atypical acinar cells, some of which are considered to appear, eventually, as acinar cell carcinomas. On the other hand, there exist nodular lesions in the human pancreas, which are similar to atypical acinar cell nodules of the rats, in the sense of nodularity, multiplicity, size, and cytological features, such as pale cytoplasm. To clarify the plausibility of the human nodular lesions as a precursor of acinar cell carcinoma of the pancreas, light and electron microscopical studies were performed, using pancreases of 115 semi-consecutive series of autopsy cases and 20 surgical cases. Multiple nodular lesions were found in 3 autopsy cases and one surgical cases. Ultrastructurally, markedly dilated cysterna of rough-surfaced endoplasmic reticulum and intracisternal granules were the most prominent characteristics of the atypical cells of the nodules. These features are neither reported in chemically induced atypical acinar cell nodules and carcinomas nor in human acinar cell carcinomas. The human lesions were considered to be of degenerative nature rather than neoplastic.     

ATYPICAL ACINAR CELL NODULES OF THE HUMAN PANCREAS

The earliest morphological evidence of altered growth potential of pancreatic acinar cells of the rats treated with carcinogen, such as azaserine, is the development of nodules of atypical acinar cells, some of which are considered to appear, eventually, as acinar cell carcinomas. On the other hand, there exist nodular lesions in the human pancreas, which are similar to atypical acinar cell nodules of the rats, in the sense of nodularity, multiplicity, size, and cytological features, such as pale cytoplasm. To clarify the plausibility of the human nodular lesions as a precursor of acinar cell carcinoma of the pancreas, light and electron microscopical studies were performed, using pancreases of 115 semi-consecutive series of autopsy cases and 20 surgical cases. Multiple nodular lesions were found in 3 autopsy cases and one surgical cases. Ultrastructurally, markedly dilated cysterna of rough-surfaced endoplasmic reticulum and intracisternal granules were the most prominent characteristics of the atypical cells of the nodules. These features are neither reported in chemically induced atypical acinar cell nodules and carcinomas nor in human acinar cell carcinomas. The human lesions were considered to be of degenerative nature rather than neoplastic.     

Dietary hyperphagia in rats: role of fat, carbohydrate, and energy content

Dietary energy, fat and carbohydrate content were varied to determine the nutritional factors responsible for hyperphagia induced by feeding rats high-fat diets. In the first experiment, rats were fed isoenergetic high-fat or high-carbohydrate diets for 2 weeks. Weight gain and energy intake were lower in rats given the high-fat diet. When some of the rats were switched to a diet that was high in fat, carbohydrate and energy, gram food intake was initially unchanged, resulting in a substantial increase in energy intake and weight gain. Energy intake gradually declined over the 4 weeks following the switch to the high-energy diet. In the second experiment, rats were fed high-fat diets that were either high or low in carbohydrate content and either high or low in energy content (kcal/g). Rats fed a high-fat diet that was high in energy and carbohydrate ate the most energy and gained the most body weight and carcass fat. In the third experiment, rats were fed high-carbohydrate diets varying in fat and cellulose content. Energy intake and body weight gain varied directly as a function of caloric density regardless of the fat or cellulose content of the diets. It is concluded that hyperphagia induced by feeding high-fat diets is not due to the high dietary fat content alone. Rather, high levels of fat, carbohydrate, and energy interact to produce overeating and obesity in rats fed high-fat diets.     

Relationship between fatty liver and subsequent development of necrosis, inflammation and fibrosis in experimental alcoholic liver disease

Rats fed a diet varying in the amount of fat, infused with ethanol, were studied to determine the relationship among diet, degree of fatty liver, and development of necrosis, inflammation, and fibrosis. Three groups of experimental animals, male Wistar rats, were fed diets containing 25% fat, 35% fat, and 32% fat with low protein. Morphologic assessment of liver injury was performed monthly by obtaining liver biopsies. The greatest degree of fatty infiltration at 1 month was seen in the high fat-low protein group, the mean fat score (3.8 +/- 0.37) was significantly higher than in the other two groups (P less than 0.05 and P less than 0.01). When the subsequent development of necrosis, inflammation, and fibrosis was related to the degree of fatty infiltration at 1 month, a significant relationship was seen between the number of animals developing these pathologic lesions and the severity of fatty liver. Our results show that the degree of fatty infiltration of the liver, influenced by the dietary intake of both fat and protein, is related to the subsequent development of necrosis, inflammation, and fibrosis in our intragastric feeding model for alcoholic liver disease.     

Diets rich in saturated fat and fructose induce anxiety and depression‐like behaviours in the rat: is there a role for lipid peroxidation?

Epidemiological studies reveal associations between obesity/metabolic syndrome and mood disorders. We assessed behavioural changes in rats fed diets enriched in fat and fructose in different proportions and correlated the observed alterations with biochemical changes induced by the diets. Three groups of rats were used as follows: control (C) animals fed regular rat chow, rats fed high‐fat diet (HF) and rats fed high‐fat and high‐fructose diet (HFHF). HF and HFHF animals were also given a 10% fructose solution as drinking water. Behavioural and biochemical parameters were determined. Anxiety was measured by the open‐field and the social interaction test. Depression‐like behaviour was evaluated by the forced swimming test. The object recognition test was utilized to assess effects on memory. Diet‐exposed animals displayed signs of anxiety in the open‐field (HF rats had reduced central time; HFHF rats had reduced number of central entries) and in the social interaction test (decreased time of interaction in HF group). In the forced swimming test, the immobility time was prolonged in the HFHF group. While different measures of anxiety scores correlated with visceral adiposity and dyslipidemia, results from both social interaction and forced swimming tests were significantly associated with lipid peroxidation, which in turn also correlated with the metabolic parameters. The experimental diets did not affect the object recognition memory. Both experimental diets induced metabolic derangements in rats and provoked similar anxiety‐ and depression‐like behaviours. Lipid peroxidation seems to play a role in translating diet‐induced metabolic alterations into behavioural disorders.     

Experimental obesity syndromes in rats: influence of diet palatability on maintenace body weights

Female rats with ventromedial hypothalamic (VMH) lesions, parasagittal hypothalamic knife cuts (KC), or dorsolateral tegmental (DLT) lesions were maintained successively on 0.2 and 0.4% quinine chow, plain chow, pellets, wet mash, and high fat diets (15–45 days each). Only VMH rats overate the 0.2% quinine diet and only KC rats underate the 0.4% quinine diet. Although DLT rats did not overeat the unadulterated chow and pellet diets, as did VMH and KC rats, all three surgical groups attained roughly comparable elevated body weight means after access to the wet mash and high fat diets. Thus, dietary manipulations clearly induce differential patterns of feeding behavior in these three obesity syndromes.     

Pancreatic Pathology,Chemically Defined Liquid Diets and Bacterial Flora in the Rat

Germfree and conventional rats on a completely defined liquid diet from weaning to adult life developed pancreatic lesions. In the germfree animals they were reversible and disappeared in the majority of this group after 5-8½ months. The lesions were encountered after ingestion of the diet for 1 month and consisted of acinar atrophy with fibrosis. The latter was largely replaced by stromal fat after 3½ months in the more vulnerable conventional rats. From 4 months of diet ingestion onward unusual focal metaplasia was observed in the pancreas of conventional animals. The metaplastic cells resembled hepatic or oxyphilic (oncocytic) epithelium and were believed to arise in the islets but the possibility of acinar or ductal origin could not be eliminated.     

Quantity not composition of dietary fats represents the dominant contributor to experimental obesity: Relevance to human pathophysiology

Plant fats are low in saturated fats but high in unsaturated fats compared to animal fats, and are supposedly less obesogenic. This study compared the obesogenic effects of plant and animal derived fatty diets in Wistar rats. Rats of each gender were divided into three dietary (standard chow (SC), high fat diet rich in animal fat (HFDaf) and a high fat diet rich in plant fat (HFDpf)) groups of ten each and fed for 17 weeks. Anthropometric, Adiposity and nutritive variables were assessed using standard methods. Comparing HFDpf to HFDaf: Abdominal circumference (AC),initial feed intaken (IFI), final feed intake(FFI), final body weight (FBW), white adipose tissue (WAT) were increased but brown adipose tissue (BAT) decreased in male rats fed with HFDpf; also, there were increased body length, IFI, FFI but decreased AC, FBW, BAT in female rats fed with HFDpf. Comparing male to female rats: Thoracic circumference, IFI, FFI, energy intake were increased while Adiposity index decreased across diet groups in male rats; the AC, FBW increased while WAT, BAT decreased in HFDpf fed group, also, BAT was increased but AC, FBW decreased in HFDaf fed group in male rats. Palatability and high feed efficiency of consumed diets were more associated with obesogenic risk than just the level of saturation. Therefore, Obesogenic effects of fatty diets in both genders is more dependent on the quantity (amount) of fatty diet consumed than the dietary fat composition alone.     

Composition of Swine Arterial Tissue

The effects of trans-fatty acid-rich and saturated acid-rich diets on the fatty acid composition and morphology of swine arterial tissues were studied. Three groups of two-month-old swine were fed either a basal diet or a basal diet containing 8.3% margarine or butter for 4 months. The most significant change observed in the fatty acid pattern was the accumulation of 18: 2ω6 and the supperssion of both 20:4ω6 and 22:4ω6 acids in the aortic tissues of swine fed either butter or margarine as compared to swine fed the basal diet. Also, ω6 metabolites were significantly decreased in swine fed diets containing butter or margarine, as compared to those fed a basal diet. The group of swine which were fed either butter or margarine containing diets suffered a significant increase in intimal thickening of the coronary artery compared to those fed the basal diet. These thickened intima were characterized by the presence of modified smooth muscle cells, lipid containing cells, and degenerated cells without stainable lipids. However, there was no significant difference in the incidence and extent of intimal thickening of the coronary arteries between swine fed either of the two fat containing diets.     

Influence of dietary fats on ultrastructure and fatty acid. Composition of swine arterial tissue

The effects of trans-fatty acid-rich and saturated acid-rich diets on the fatty acid composition and morphology of swine arterial tissues were studied. Three groups of two-month-old swine were fed either a basal diet or a basal diet containing 8.3% margarine or butter for 4 months. The most significant change observed in the fatty acid pattern was the accumulation of 18 : 2 omega 6 and the suppression of both 20 : 4 omega 6 and 22 : 4 omega 6 acids in the aortic tissues of swine fed either butter or margarine as compared to swine fed the basal diet. Also, omega 6 metabolites were significantly decreased in swine fed diets containing butter or margarine, as compared to those fed a basal diet. The group of swine which were fed either butter or margarine containing diets suffered a significant increase in intimal thickening of the coronary artery compared to those fed the basal diet. These thickened intima were characterized by the presence of modified smooth muscle cells, lipid containing cells, and degenerated cells without stainable lipids. However, there was no significant difference in the incidence and extent of intimal thickening of the coronary arteries between swine fed either of the two fat containing diets.     

Cingulate lesions and behavioral adaptation to amino acid imbalanced diets

The effect of anterior cingulate cortex lesions on dietary intake and adaptation of disproportionate amounts of amino acids was examined. Rats with bilateral electrolytic lesions in the anterior cingulate cortex and sham-operated rats were fed, in turn, amino acid basal, imbalanced or devoid diets involving threonine and isoleucine as the growth limiting amino acids, and then a low protein (6% casein) followed by a high protein (75% casein) diet. Lesions of the anterior cingulate cortex did not prevent the initial depression in food intake of the amino acid imbalanced diets, but shortened the duration of anorexia associated with dietary amino acid imbalances. Cingulate lesions did not influence the food intake of rats fed amino acid devoid diets. When switched from a low protein to a high protein diet, animals bearing lesions and sham-operated controls reduced markedly their initial food intake and adapted to the high protein diet in similar manner. It was concluded that the initial food intake depression associated with a dietary amino acid imbalance is a direct response to postingestive cues which influence food intake. Moreover, that the difference in adaptive intakes of the cingulate cortex lesioned animals who ingested a diet of imbalanced amino acids or of high protein, indicates that separate mechanisms act to control food intake of animals fed diets containing imbalanced amino acid mixtures or diets with excessive amounts of protein.