Dietary α-Lactalbumin protects against thioacetamide-induced liver cirrhosis by maintaining gut-liver axis function in rats.

We tested the hypothesis that α-lactalbumin inhibits the disruption of intestinal barrier function and liver cirrhosis by restoring gut-liver axis function in thioacetamide (TAA) -treated rats. Rat diets were supplemented with α-lactalbumin replacing 50% of dietary protein. After consuming α-lactalbumin for one week, rats were intraperitoneally injected with TAA twice a week for 14 weeks. The α-lactalbumin-enriched diet significantly inhibited the elevation of plasma alanine aminotransferase, aspartate aminotransferase, and hyaluronic acids. The supplement significantly reduced plasma lipopolysaccharide levels and increased occludin mRNA level. Hepatic fibrosis and regenerative nodules was developed and intestinal villi were shortened by TAA; α-Lactalbumin attenuated these histopathological changes. These results indicated that α-lactalbumin improved intestinal barrier function, suppressing endotoxin levels. These data also suggested that α-lactalbumin ameliorated the impairment of the gut-liver axis by TAA, inhibiting the development of liver cirrhosis.

Modified citrus pectin stops progression of liver fibrosis by inhibiting galectin-3 and inducing apoptosis of stellate cells.

Modified citrus pectin (MCP) is a pH modified form of the dietary soluble citrus peel fiber known as pectin. The current study aims at testing its effect on liver fibrosis progression. Rats were injected with CCl4 (1 mL/kg, 40% v/v, i.p., twice a week for 8 weeks). Concurrently, MCP (400 or 1200 mg/kg) was administered daily in drinking water from the first week in groups I and II (prophylactic model) and in the beginning of week 5 in groups III and IV (therapeutic model). Liver function biomarkers (ATL, AST, and ALP), fibrosis markers (laminin and hyaluronic acid), and antioxidant biomarkers (reduced glutathione (GSH) and superoxide dismutase (SOD)) were measured. Stained liver sections were scored for fibrosis and necroinflammation. Additionally, expression of galectin-3 (Gal-3), α-smooth muscle actin (SMA), tissue inhibitor metalloproteinase (TIMP)-1, collagen (Col)1A1, caspase (Cas)-3, and apoptosis related factor (FAS) were assigned. Modified pectin late administration significantly (p < 0.05) decreased malondialdehyde (MDA), TIMP-1, Col1A1, α-SMA, and Gal-3 levels and increased levels of FAS, Cas-3, GSH, and SOD. It also decreased percentage of fibrosis and necroinflammation significantly (p < 0.05). It can be concluded that MCP can attenuate liver fibrosis through an antioxidant effect, inhibition of Gal-3 mediated hepatic stellate cells activation, and induction of apoptosis.

Dietary supplementation with monounsaturated and long-chain polyunsaturated fatty acids influences the liver structural recovery and hepatocyte binuclearity in female Wistar rats in experimental cirrhosis induced by thioacetamide.

UNLABELLED: Oral administration of 300 mg/l thioacetamide (TAA) for 4 months causes hepatic lesions comparable to those described in alcoholic liver cirrhosis in humans and associated protein-energy malnutrition. In this sense, direct supplementation with monounsaturated fatty acids (MUFAs) and/or polyunsaturated fatty acids (PUFAs) might provide an advantage in the correction of the fatty acid deficiency in these patients. PUFAs are essential components of cell membranes maintaining its fluidity and function, important energy sources, and precursors of eicosanoids. Moreover, these fatty acids also modulate gene transcription, mRNA stability, and cellular differentiation. METHODS: Fifty-four female Wistar rats (Interfauna Ibérica, Barcelona, Spain) weighing 110-120 g were used in this study. The animals were divided into two groups: one group was treated with 300 mg/l TAA dissolved in drinking water during 4 months, and the other group, which served as a control, was given water without TAA. To evaluate the changes induced by the administration of TAA for 4 months, TAA-treated (n = 7) and control animals (n = 5) were killed. Then, the TAA treatment was stopped and the rest of the animals in both TAA and control groups were divided into three experimental groups and three control groups which received for 2 weeks different type of diets. Using the TAA-induced liver cirrhosis model in rats, we analysed the effects of dietary supplementation with MUFAs and PUFAs on binuclearity and ultrastructure of hepatocytes. After TAA-induced cirrhosis, we analysed whether dietary supplementation with fatty acids may restore the normal percentage of binucleated cells, as well as the ultrastructure, nuclear area, and nuclear/cytoplasm index of hepatocytes. RESULTS: Treatment with TAA causes cirrhosis characterized by the appearance of parenchyma nodules and fibrous septae, as well as qualitative and quantitative alterations in liver and plasma lipids. Our results indicate that dietary MUFAs support hepatocyte recovery regarding its ultrastructural and morphometric values. However, PUFAs-enriched diets (n-3 and n-3 + n-6) do not correct hepatomegaly, fibrosis or lipid accumulation. Thus, dietary PUFAs do not enhance hepatocyte recovery from morphological and ultrastructural alterations. CONCLUSIONS: In our experimental model of cirrhosis, dietary supplementation with a high proportion of long-chain PUFAs (n-3 or n-6) negatively influences liver recovery. This negative effect was likely due to the increased susceptibility of cell membranes to lipid peroxidation, together with an alteration in lipid metabolism.

Effect of iron overload and dietary fat on indices of oxidative stress and hepatic fibrogenesis in rats.

BACKGROUND/AIMS: Oxidative stress is presumed to play an important role in hepatic fibrogenesis. Diets high in polyunsaturated fatty acids (PUFA) enhance fibrosis and have been associated with increased oxidative damage in some models of liver injury. The aim of this study was to determine the effects of dietary fat of varying PUFA content on iron-induced oxidative stress and fibrosis. METHODS: Rats were given parenteral iron and diets supplemented with coconut oil, safflower oil or menhaden oil. RESULTS: Hepatic iron overload was associated with induction of heme oxygenase-1, a sensitive indicator of oxidative stress, and with modest increases in hydroxyproline and procollagen I mRNA levels without histologically evident fibrosis, all of which were unaffected by dietary fat. In addition, iron loading was associated with increases in cysteine, gamma-glutamylcysteine and glutathione. Dietary fat brought about the expected alterations in peroxidizability, but did not alter indices of oxidative damage. CONCLUSION: These data highlight the distinction between oxidative stress and oxidative damage and suggest that the former is not sufficient to elicit overt fibrosis. Furthermore, while hepatic iron overload leads to oxidative stress, there is an associated upregulation of antioxidant defenses involving thiol metabolism that may be a critical factor limiting the accumulation of oxidative damage.

Effect of branched-chain amino acids on the composition and cytolytic activity of liver-associated lymphocytes in rats.

BACKGROUND: Although branched-chain aminoacids (BCAA) are reported to be effective in prolongation of the mean survival time of patients with liver cirrhosis, it is not clear whether BCAA could influence the immune function in those patients. METHODS: Branched-chain amino acids were given as a supplement to carbon tetrachloride-induced cirrhotic rats, and an aminogram of the liver and kinetics of liver-associated lymphocytes (LAL) were then analysed. RESULTS: Liver cirrhosis was established at the 12th week, and glutathione S-transferase placental form (GST-P)-positive lesions, which are known to be pre-neoplastic lesions, occupied 1.72+/-0.84% of the liver at the 16th week in the controls. At this time the LAL showed an increase in the number of CD5-, CD8- and CD18-positive cells and augmentation of lectin-dependent cellular cytotoxicity (LDCC) activity. Furthermore, supplementation of BCAA increased the number of LAL, especially CD8-positive cells and natural killer cells, and augmented LDCC activity of LAL at the 16th week. The number of LAL was positively correlated with the valine concentration in the plasma and liver, and the area of GST-P-positive lesions tended to be decreased in the BCAA group. CONCLUSION: These findings suggest that BCAA administration has stimulatory effects on the local immune systems of the liver, which may have a potential to inhibit hepatocarcinogenesis. Moreover, among all amino acids valine might be an important amino acid for enhancing the immune function of LAL.

Oral supplementation with branched-chain amino acids improves survival rate of rats with carbon tetrachloride-induced liver cirrhosis.

We investigated whether supplementation with branched-chain amino acids (BCAA) improves survival of rats with carbon tetrachloride (CCl4) -induced cirrhosis. Liver cirrhosis was induced in 40 male Sprague-Dawley rats by administering CCl4 for 15 weeks. Twenty rats each were then assigned to the control and BCAA group and fed a casein diet or a BCAA-supplemented casein diet, respectively, for an additional 17 weeks with repeated injections of CCl4. No significant difference occurred in either mean energy or nitrogen intake or in body or liver weight between the two groups. BCAA-supplementation significantly preserved plasma albumin concentrations (P < 0.05) and inhibited significantly the occurrence of ascites and hyperammonemia (P < 0.05). The survival rate was significantly higher in the BCAA group (P=0.03), while no significant difference was found in liver histology between the groups. These results suggest that BCAA improved survival of rats with CCl4-induced cirrhosis by preventing hypoalbuminemia and hyperammonemia without directly reducing hepatic necrosis and fibrosis.

Dietary nucleotides correct plasma and liver microsomal fatty acid alterations in rats with liver cirrhosis induced by oral intake of thioacetamide.

BACKGROUND/AIMS: Dietary nucleotides modulate a number of metabolic processes, including long-chain polyunsaturated fatty acid metabolism. In this study, we evaluated the effect of dietary nucleotides on plasma and liver microsomal fatty acid profiles in a rat model of liver cirrhosis induced by oral intake of thioacetamide. METHODS: Fifty-four female Wistar rats were assigned to one of the following groups: rats in the thioacetamide group (n=45) were given 300 mg thioacetamide/l in their drinking water for 4 months, and rats in the control group (n=9) received water during the same period. After 4 months of treatment, 9 rats in each group were killed. The remaining rats in the thioacetamide group were divided into two new groups, and the animals in each were allowed to recover for 1 or 2 weeks on either a nucleotide-free diet or the same diet supplemented with 50 mg of each of the following: AMP, GMP, CMP, IMP and UMP per 100 g diet. RESULTS: Saturated (mainly stearic acid), monounsaturated, and n-6 long-chain polyunsaturated fatty acids (mainly arachidonic acid), and also the unsaturation index decreased in plasma of rats with experimental cirrhosis. Administration of the diet supplemented with nucleotides to thioacetamide-treated rats corrected plasma levels of saturated, n-6 long-chain polyunsaturated fatty acids and the unsaturation index. In liver microsomes, the cirrhotic rats showed lower levels of protein and higher levels of palmitic, oleic, linoleic and arachidonic acids. Protein concentrations and levels of all the above-mentioned fatty acids were corrected with the nucleotide-enriched diet. CONCLUSIONS: Dietary nucleotides contribute to correcting plasma and liver microsomal fatty acid alterations in rats with liver cirrhosis induced by chronic oral administration of thioacetamide.

Steatosis and collagen content in experimental liver cirrhosis are affected by dietary monounsaturated and polyunsaturated fatty acids.

BACKGROUND AND METHODS: We used thioacetamide administered orally to induce cirrhosis in rats, and after these had recovered for 1 and 2 weeks we examined the effects of dietary supplementation with monounsaturated and n-3 polyunsaturated fatty acids, or with a combination of n-3 and n-6 polyunsaturated fatty acids, on the extent of steatosis and collagen content in the liver. RESULTS: Nodular cirrhosis, increased collagen content, and lipid accumulation were established after 4 months of treatment with thioacetamide. When the animals were fed a diet rich in oleic acid for 2 weeks, the steatosis and fibrosis decreased. Supplementation with n-3 polyunsaturated fatty acids favored reductions in collagen content but did not reduce the fat accumulation. With a diet supplemented with a mixture of n-3 and n-6 fatty acids we found no reduction in either lipid accumulation or collagen content. CONCLUSIONS: Fibrosis and steatosis may be influenced by dietary fat, and monounsaturated fat appears to influence favorably the histologic recovery of the damaged liver.

Dietary long-chain polyunsaturated fatty acids influence the recovery of thioacetamide-induced liver cirrhosis in rats.

BACKGROUND: The aim of this study was to evaluate the dietary supplementation with omega-3 and omega-6 long-chain polyunsaturated fatty acids on the fatty acid composition of plasma and red blood cell membranes in rats with thioacetamide-induced liver cirrhosis. METHODS: Thirty-eight female Wistar rats were given 300 mg thioacetamide/L in drinking water for 4 months to induce the experimental liver cirrhosis. Sixteen rats were used as controls. After treatment with thioacetamide, nine rats of each group were killed. Then, thioacetamide-treated rats were divided into three new groups, each receiving a different diet for 2 weeks: a semipurified diet (n = 9), the same diet supplemented with omega-3 long-chain polyunsaturated fatty acids (n = 10), or the same semipurified diet supplemented with omega-3 and omega-6 long-chain polyunsaturated fatty acids simultaneously (n = 10). The remaining control rats were fed the semipurified diet. Liver histology and plasma and erythrocyte fatty acid composition were assessed. RESULTS: An apparent improvement of the histological damage took place in the rats fed the omega-3+ omega-6-supplemented diet. The diet supplemented with polyunsaturated fatty acids of the omega-3 series induced increases in the omega-3 long-chain polyunsaturated fatty acid levels in total plasma lipids, plasma lipid fractions and in erythrocyte phospholipids, and decreases in omega-6 long-chain polyunsaturated fatty acids in erythrocyte phospholipids during the recovery of rats with thioacetamide-induced liver cirrhosis. The administration of the diet supplemented with both omega-3 and omega-6 long-chain polyunsaturated fatty acids contributed to increase the levels of total plasma saturated, monounsaturated, and omega-6 long-chain polyunsaturated fatty acids from cirrhotic rats. CONCLUSION: We conclude that the simultaneous supply of long-chain fatty acids of the omega-3 and the omega-6 series can be beneficial to improve the fatty acid status of this experimental model of liver cirrhosis.