concanavalin-a and Choline-Deficiency

To determine the immune-modulatory and the hepatoprotective effects of oral administration of two soy extracts in immune mediated liver injury and non-alcoholic steatohepatitis (NASH).. Two soy extracts, M1 and OS, were orally administered to mice with concanavalin A (ConA) immune-mediated hepatitis, to high-fat diet (HFD) mice and to methionine and choline reduced diet combined with HFD mice. Animals were followed for disease and immune biomarkers.. Oral administration of OS and M1 had an additive effect in alleviating ConA hepatitis manifested by a decrease in alanine aminotransferase and aspartate aminotransferase serum levels. Oral administration of the OS and M1 soy derived fractions, ameliorated liver injury in the high fat diet model of NASH, manifested by a decrease in hepatic triglyceride levels, improvement in liver histology, decreased serum cholesterol and triglycerides and improved insulin resistance. In the methionine and choline reduced diet combined with the high fat diet model, we noted a decrease in hepatic triglycerides and improvement in blood glucose levels and liver histology. The effects were associated with reduced serum tumor necrosis factor alpha and alteration of regulatory T cell distribution.. Oral administration of the combination of OS and M1 soy derived extracts exerted an adjuvant effect in the gut-immune system, altering the distribution of regulatory T cells, and alleviating immune mediated liver injury, hyperlipidemia and insulin resistance.

Topics: Administration, Oral; Animals; Biomarkers; Blood Glucose; Chemical and Drug Induced Liver Injury; Choline Deficiency; Concanavalin A; Cytoprotection; Diet, High-Fat; Disease Models, Animal; Glycine max; Immunologic Factors; Insulin; Insulin Resistance; Lipids; Liver; Metabolic Syndrome; Methionine; Mice; Non-alcoholic Fatty Liver Disease; Plant Extracts; T-Lymphocytes, Regulatory; Time Factors; Tumor Necrosis Factor-alpha

Steatohepatitis enhances the severity of liver injury caused by acute inflammation. The purpose of this study was to test the hypothesis that fatty liver due to chronic choline-deficient diet exacerbates concanavalin A (ConA)-induced liver hepatitis, which is predominantly facilitated by T cells. Male C57BL/6 mice were fed either control choline-sufficient diet (CSD) or choline-deficient diet (CDD) for 6 weeks before ConA administration. Mice were sacrificed 3, 9, and 24 hours after ConA injection. Liver injury measured by aspartate aminotransferase (AST), alanine aminotransferase (ALT), pathology, and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) staining was minimal in mice fed either diet before ConA exposure. However, ConA-induced liver injury was significantly greater in CDD-fed mice compared with control-fed mice. Liver cytokines were assessed by quantitative real-time polymerase chain reaction (PCR). The expression of T helper (Th) 1 cytokines tumor necrosis factor alpha (TNF-alpha), interleukin 12 (IL-12), and interferon gamma (IFN-gamma) were dramatically elevated after ConA in CDD-fed mice compared with control-fed mice. CDD also enhanced ConA-induced STAT4 activation, but not STAT6. Notably, regulators of T-cell differentiation were strongly shifted toward a predominant Th1 profile. T-bet, regulator of the Th1 response, was up-regulated in CDD-fed mice, whereas Th2 regulator GATA-3 was significantly suppressed in CDD-fed mice after ConA. Moreover, the expression of suppressor of cytokine signaling (SOCS)-1, SOCS-3, and repressor of GATA-3 (ROG) favored a predominant Th1 cytokine response in CDD-fed mice. In conclusion, these data support the hypothesis that hepatosteatosis caused by CDD is associated with more severe ConA-induced hepatitis due to a predominant shift toward Th1 response.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Blotting, Western; Cell Differentiation; Chemical and Drug Induced Liver Injury; Choline Deficiency; Concanavalin A; Cytokines; Disease Models, Animal; Disease Progression; Fatty Liver; GATA3 Transcription Factor; Gene Expression; Male; Mice; Mice, Inbred C57BL; Polymerase Chain Reaction; RNA, Messenger; T-Box Domain Proteins; T-Lymphocytes; Th1 Cells; Transcription Factors

The effect of variations in culture media, mitogen dose, harvest time, 2-mercaptoethanol (2-ME) addition and length of [3H]thymidine pulse on the concanavalin A (Con A)-stimulated splenic lymphocyte transformation response of male, weanling Sprague-Dawley rats maintained on a control (C), folacin-deficient (F) or marginal methionine-choline (M/C) diet was examined. Splenocytes cultured in minimal essential medium reached an optimal response on day 4. The response of F rats was significantly lower than C rats on day 3 and day 4. The response of M/C rats were lower on day 3. The addition of 2-ME to the culture medium shifted the cell cycle kinetics toward and earlier peak response time (day 2), and significant differences among groups were seen only under suboptimal conditions. Cells cultured in Medium 199 had a low transformation response, which reached peak stimulation on day 5. The response of F and M/C rats was significantly lower than C animals on days 3 and 4. In contrast, splenocytes cultured in medium 199 + 2-ME reached optimal stimulation on day 2, with no significantly differences between groups. No effect on cell viability was seen from 2-ME, but it did accelerate cell cycle kinetics and reversed the normal age-induced immunosuppression seen in C animals.

Topics: Animals; Cells, Cultured; Choline Deficiency; Concanavalin A; Culture Media; Dose-Response Relationship, Drug; Folic Acid Deficiency; Lymph Nodes; Lymphocyte Activation; Male; Mercaptoethanol; Methionine; Rats; Rats, Inbred Strains; Spleen; Thymidine; Thymus Gland

We attempted to identify variants of gamma-glutamyl transferase (GGT) which may differ in their affinity for Concanavalin A (Con A) in extracts of fetal, regenerating, neoplastic and normal adult rat liver. GGT activity was low in normal liver and did not increase after partial hepatectomy. Feeding a choline-deficient diet containing 0.05% ethionine caused a large increase in hepatic GGT activity which persisted throughout the feeding period of 23 weeks. Practically all of the GGT contained in extracts prepared with Triton X-100 from fetal, regenerating, pre-neoplastic and neoplastic rat liver as well as that from normal adult liver bound to Con A-Sepharose and was eluted from the columns with methyl-alpha-D-mannopyranoside. In contrast, GGT from bovine kidney extracts prepared in an identical way did not bind to Con A-Sepharose. We conclude that if variants of rat liver GGT exist, they cannot be reliable separated by simple affinity chromatography on Con A-Sepharose of Agarose columns.

Topics: Animals; Choline Deficiency; Chromatography, Affinity; Concanavalin A; Female; Fetus; gamma-Glutamyltransferase; Isoenzymes; Liver; Liver Neoplasms; Liver Regeneration; Male; Neoplasms, Experimental; Pregnancy; Rats; Rats, Inbred Strains