CL Diet Research Articles

Cenicriviroc Suppresses and Reverses Steatohepatitis by Regulating Macrophage Infiltration and M2 Polarization in Mice.

The inhibition of hepatic macrophage and Kupfer cell recruitment and activation is a potential strategy for treating insulin resistance and nonalcoholic steatohepatitis (NASH). Cenicriviroc (CVC), a dual C-C chemokine receptor 2 (CCR2) and CCR5 antagonist, has shown antifibrotic activity in murine models of NASH and has been evaluated in clinical trials on patients with NASH. This study investigated the effects of CVC on macrophage infiltration and polarization in a lipotoxic model of NASH. C57BL/6 mice were fed a high-cholesterol, high-fat (CL) diet or a CL diet containing 0.015% CVC (CL + CVC) for 12 weeks. Macrophage recruitment and activation were assayed by immunohistochemistry and flow cytometry. CVC supplementation attenuated excessive hepatic lipid accumulation and peroxidation and alleviated glucose intolerance and hyperinsulinemia in the mice that were fed the CL diet. Flow cytometry analysis revealed that compared with the CL group, mice fed the CL + CVC diet had fewer M1-like macrophages, more M2-like macrophages, and fewer T cell counts, indicating that CVC caused an M2-dominant shift of macrophages in the liver. Similarly, CVC decreased lipopolysaccharide-stimulated M1-like macrophage activation, whereas it increased interleukin-4-induced M2-type macrophage polarization in vitro. In addition, CVC attenuated hepatic fibrosis by repressing hepatic stellate cell activation. Lastly, CVC reversed insulin resistance as well as steatosis, inflammation, and fibrosis of the liver in mice with pre-existing NASH. In conclusion, CVC prevented and reversed hepatic steatosis, insulin resistance, inflammation, and fibrogenesis in the liver of NASH mice via M2 macrophage polarization.

Chemokine receptor CCR1 regulates macrophage activation through mTORC1 signaling in nonalcoholic steatohepatitis

Background and aimsChemokine (CC motif) receptor 1 (CCR1) promotes liver fibrosis in mice. However, its effects on nonalcoholic steatohepatitis (NASH) remain unclear. Therefore, the present study aimed to investigate the role of CCR1 in the progression of NASH. MethodsHuman serum and liver tissues were obtained from patients with NASH and controls. Systemic (Ccr1−/−) and liver macrophage-knockout Ccr1 (Ccr1LKD) mice were fed a high-cholesterol and high-fat (CL) diet for 12 weeks or a methionine/choline-deficient (MCD) diet for 4 weeks. BX471 was used to pharmacologically inhibit CCR1 in CL-fed mice. ResultsCCR1 was significantly upregulated in liver samples from patients with NASH and in animal models of dietary-induced NASH. In the livers of mice fed a CL diet for 12 weeks, the CCR1 protein colocalized with F4/80+ macrophages rather than with hepatic stellate cells. Compared to their wild-type littermates, Ccr1−/− mice fed with the CL or MCD diet showed inhibition of NASH-associated hepatic steatosis, inflammation, and fibrosis. Mechanistically, Ccr1 deficiency suppressed macrophage infiltration and activation by attenuating the mechanistic target of rapamycin complex 1 (mTORC1) signaling. Similar results were observed in Ccr1LKD mice administered the CL diet. Moreover, CCR1 inhibition by BX471 effectively suppressed NASH progression in CL-fed mice. ConclusionsCcr1 deficiency mitigated macrophage activity by inhibiting mTORC1 signaling, thereby preventing the development of NASH. Notably, the CCR1 inhibitor BX471 protected against NASH. These findings would help in developing novel strategies for the treatment of NASH.

Broiler meat fatty acids composition, lipid metabolism, and oxidative stability parameters as affected by cranberry leaves and walnut meal supplemented diets

A randomized complete block with a 2 × 3 factorial arrangement was used to design a nutrition experiment conducted for the evaluation of the relation between walnut meal (WM—6% inclusion rate) and cranberry leaves (CL—1% and 2% inclusion rate) supplements and their effects on tissue lipid profile, lipid metabolism indices and oxidative stability of meat. Semi-intensive system conditions were simulated for 240 Ross 308 broilers and the animals were reared on permanent shave litter in boxes of 3 m2 (40 broilers / each group, housed in a single box). The current study results showed that the diets enriched in linolenic acid (LNA) (WM diets) led to broilers meat enriched in LNA, but the synthesis of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) was stimulated when the diets were supplemented with a natural antioxidants source (CL diets). The CL diet also exhibited the most powerful effect in counteracting the oxidative processes of meat.

Effect of tropical forage species in silvopastoral arrangements on methane production and in vitro fermentation parameters in a RUSITEC system

Background: Supplementation of grazing cattle with native and naturalized forages using silvopastoral systems has been suggested as an affordable strategy to reduce methane production and improve nutrition, diminishing the environmental impact of cattle production. Objective: To evaluate the effect of three tropical forage species in a silvopastoral arrangement on methane production and fermentation parameters using an in vitro ruminal simulation system (RUSITEC). Methods: Four diets were evaluated. The control treatment was a basal diet of colosuana grass (COL; Bothriochloa pertusa), while the other diets consisted of 70% COL complemented with 30% shrub forage from either Leucaena leucocephala (CL), Guazuma ulmifolia (CG), or Crescentia cujete (CT). A randomized complete block design with repeated measurements over time was used. Results: The inclusion of shrub forage did not affect pH, organic matter degradation (OMD) or volatile fatty acids (VFA). The inclusion of shrub forage affected the degradation of structural components. The concentration of N-NH3 increased in the CL diet compared to COL (p<0.05). In general, methane production in terms of mL/day, mL/g DMi, mL/g DMd, and mL/gOMd was reduced for CL compared to COL (p<0.05). Conclusions: Based on these results, inclusion of Leucaena leucocephala, Guazuma ulmifolia or Crescentia cujete on B. pertusa-based diets improves ruminal fermentation parameters and reduces in vitro methane production.

C[sbnd]C chemokine ligand 3 deficiency ameliorates diet-induced steatohepatitis by regulating liver macrophage recruitment and M1/M2 status in mice

Background and aimsThe global prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing. Chemokines and their receptors have potential as therapeutic targets of NAFLD. We investigated the role of CC chemokine ligand 3 (CCL3) in the development of murine and human NAFLD. MethodsCCL3-knockout mice (CCL3−/−) and littermate CCL3 wild-type control mice (WT) were fed a high-cholesterol and high-fat (CL) diet for 16 weeks to induce NAFLD. We investigated the impact of CCL3 gene deletion in bone marrow cells and leptin-deficient ob/ob mice on CL diet-induced steatohepatitis. We assayed the serum CCL3 levels in 36 patients with biopsy-proven NAFLD and nine healthy control subjects. ResultsCompared with normal chow (NC), the CL diet induced steatohepatitis and hepatic fibrosis and elevated the plasma CCL3 level. In the liver, CCL3 protein colocalized with F4/80+ macrophages, especially CD11c+ M1-like macrophages, rather than other cell types. CCL3−/− attenuated CL diet-induced steatohepatitis and fibrosis associated with M2-dominant liver macrophages compared with the WT. The reconstitution of bone marrow (BM) cells from CCL3−/− attenuated steatohepatitis in WT mice fed a CL diet. Furthermore, crossing CCL3−/− onto the ob/ob background prevented CL diet-induced NAFLD in ob/ob mice, which was associated with a lesser inflammatory phenotype of liver macrophages. Also, the serum and hepatic levels of CCL3 were significantly increased in patients with non-alcoholic steatohepatitis (NASH) compared to those with simple fatty liver (NAFL) and healthy subjects. ConclusionOur data indicate that CCL3 facilitates macrophage infiltration into the liver and M1 polarization in the progression of steatohepatitis and highlight the need for further studies to determine the effect of CCL3-CCR1 and -CCR5 signaling blockade on the treatment of NAFLD.

Response of plantain (Plantago lanceolata L.) supplementation on nutritional, endo-parasitic, and endocrine status in lambs

The aim of the present study was to assess the effects of plantain herb (Plantago lanceolata L.) supplementation on growth, plasma metabolites, liver enzymatic activity, hormonal status, gastrointestinal parasites, and carcass characteristics of lambs. A total of 24 lambs, aged 6 months weighing 8.0 ± 0.5kg were randomly allocated to one of two dietary treatments: (1) CL diet-roadside grass and concentrate mixture; (2) PL diet-CL diet + 5% fresh plantain supplementation on a DM basis. The PL diet group exhibited 23% higher (P = 0.01) average daily gain and 15% improved (P = 0.03) feed conversion efficiency. Circulating cholesterol concentrations were suppressed by 9% (P = 0.03), and liver enzyme activity was improved by 5-25% (P < 0.05) in the lamb fed PL diet, compared with CL diet only. The inclusion of plantain in the diet was highly effective at suppressing the parasites, Paramphistomum spp. (P = 0.003) and coccidial parasites (P = 0.04), but not stomach worms. Moreover, plantain supplementation increased growth hormone and insulin concentrations in plasma level, whereas decreased carcass fat by 32.7%. Therefore, supplementation of the lambs' diet with plantain showed some beneficial effects on productivity and parasitic infection, while it led to a leaner carcass.

Blueberry and cranberry anthocyanin extracts reduce bodyweight and modulate gut microbiota in C57BL/6J mice fed with a high-fat diet.

Blueberry and cranberry are rich in anthocyanins. The present study was to investigate the effects of anthocyanin extracts from blueberry and cranberry on body weight and gut microbiota. C57BL/6J Mice were divided into six groups (n = 9 each) fed one of six diets namely low-fat diet (LFD), high-fat diet (HFD), HFD with the addition of 1% blueberry extract (BL), 2% blueberry extract (BH), 1% cranberry extract (CL), and 2% cranberry extract (CH), respectively. Feeding BL and BH diets significantly decreased body weight gain by 20-23%, total adipose tissue weight by 18-20%, and total liver lipids by 16-18% compared with feeding HFD. Feeding CH diet but not CL diet reduced the body weight by 27%, accompanied by a significant reduction of total plasma cholesterol by 25% and tumor necrosis factor alpha(TNF-α) by 38%. The metagenomic analysis showed that the supplementation of blueberry and cranberry anthocyanin extracts reduced plasma lipopolysaccharide concentration, accompanied by a reduction in the relative abundance of Rikenella and Rikenellaceae. Dietary supplementation of berry anthocyanin extracts promoted the growth of Lachnoclostridium, Roseburia, and Clostridium_innocuum_group in genus level, leading to a greater production of fecal short-chain fatty acids (SCFA). It was concluded that both berry anthocyanins could manage the body weight and favorably modulate the gut microbiota at least in mice.

Lactobacillus pentosus strain S-PT84 improves steatohepatitis by maintaining gut permeability.

Intestinal mucosal barrier dysfunction is closely related to the pathogenesis of nonalcoholic steatohepatitis (NASH). Gut immunity has been recently demonstrated to regulate gut barrier function. The Lactobacillus pentosus strain S-PT84 activates helper T cells and natural killer/natural killer T cells. In this study, we examined the effect of S-PT84 on NASH progression induced by high-cholesterol/high-fat diet (CL), focusing on the immune responses involved in gut barrier function. C57BL/6 mice were fed a normal chow or CL diet with or without 1 × 1010 S-PT84 for 22 weeks. S-PT84 administration improved hepatic steatosis by decreasing triglyceride and free fatty acid levels by 34% and 37%, respectively. Furthermore, S-PT84 inhibited the development of hepatic inflammation and fibrosis, suppressed F4/80+ macrophage/Kupffer cell infiltration, and reduced liver hydroxyproline content. Administration of S-PT84 alleviated hyperinsulinemia and enhanced hepatic insulin signalling. Compared with mice fed CL diet, mice fed CL+S-PT84 had 71% more CD11c-CD206+ M2 macrophages, resulting in a significantly decreased M1/M2 macrophage ratio in the liver. Moreover, S-PT84 inhibited the CL diet-mediated increase in intestinal permeability. Additionally, S-PT84 reduced the recruitment of interleukin-17-producing T cells and increased the levels of intestinal tight junction proteins, including zonula occludens-1, occludin, claudin-3, and claudin-7. In conclusion, our findings suggest that S-PT84 attenuates diet-induced insulin resistance and subsequent NASH development by maintaining gut permeability. Thus, S-PT84 represents a feasible approach to prevent the development of NASH.

DPP-4 Inhibition with Anagliptin Reduces Lipotoxicity-Induced Insulin Resistance and Steatohepatitis in Male Mice

Excessive hepatic lipid accumulation drives the innate immune system and aggravates insulin resistance, hepatic inflammation, and fibrogenesis, leading to nonalcoholic steatohepatitis (NASH). Dipeptidyl peptidase-4 (DPP-4) regulates glucose metabolism and is expressed in many different cell types, including the cells of the immune system. In addition, DPP-4 may be involved in macrophage-mediated inflammation and insulin resistance. This study investigated the effects of anagliptin (Ana), an inhibitor of DPP-4, on macrophage polarity and phenotype in the livers of mice with steatohepatitis. We investigated the effects of Ana on steatohepatitis induced via a high-cholesterol high-fat (CL) diet or a choline-deficient L-amino acid-defined, high-fat (CDAHF) diet. DPP-4 activity, liver histology, and insulin sensitivity were evaluated, and liver DPP-4+ macrophages were quantified using fluorescence-activated cell sorting (FACS). Liver and plasma DPP-4 activity increased significantly in mice on both diets. FACS revealed that, compared with chow-fed mice, the CL-fed mice exhibited a significant increase in the proportion of DPP-4+ liver macrophages, particularly the M1-type macrophages. Ana decreased hepatic lipid and M1 macrophage accumulation and stimulated M2 macrophage accumulation in the liver, thereby attenuating insulin resistance, steatohepatitis, and fibrosis. Importantly, Ana alleviated hepatic fibrosis and steatohepatitis in mice fed CL diet and CDAHF diet. Using Ana to inhibit DPP-4 reduced lipotoxicity-induced hepatic insulin resistance through regulating the M1/M2 macrophage status.

Effect of herbal supplementation on growth, immunity, rumen histology, serum antioxidants and meat quality of sheep

There is a wide range of feed additives deliberately designed to be used in sheep diets that can improve production performance. Whereas herbal supplementation is gaining popularity not only for improving sheep productivity and mutton quality but also for safe application without any harmful residual effects. The present study was designed to investigate the effect of plantain (Plantago lanceolata L.) and/or garlic leaf (Allium sativum) dietary supplementation on growth performance, immunity, rumen histology, serum antioxidants and meat quality of sheep. The experiment consisted of a completely randomized design with 32 one-year-old sheep (initial mean live weight 9 ± 0.2 kg) allocated to four groups (8 sheep per group). Rice straw and concentrates-based total mixed ration pellets (2390 kcal/kg DM, CP = 15.1%) were offered as a control diet (CL diet). Herbal treatment diets included (i) CL diet + 10 g DM of plantain herb (PL diet), (ii) CL diet + 10 g DM of garlic leaf (GL diet) and (iii) CL diet + 5 g DM of PL and 5 g DM of GL (PG diet). Compared with the CL diet group, the live weight gain and feed conversion ratio were 18% to 26% and 13% to 20% higher in herbal-supplemented groups, respectively. Moreover, the herbal-supplemented groups, especially the PL diet group had higher serum immunoglobulin concentration, antioxidant capacity and rumen papillae size compared to the control. Besides, the lowest caul fat and pelvic fat levels were observed in the PL diet group followed by PG, GL and CL diet groups. In addition, lower mutton ether extract and saturated fatty acid along with higher polyunsaturated fatty acid levels were found in all herbal-supplemented groups. In conclusion, dietary supplementation with PL and/or GL might be used as an alternative in sheep to promote growth, health status and lean mutton production.

1865-P: Dual Chemokine Receptor CCR2/CCR5 Antagonist Cenicriviroc Ameliorates Lipotoxicity-Induced Insulin Resistance and NASH in Mice

Hepatic lipid accumulation drives innate immunity with recruitment of bone marrow-derived macrophages and activation of liver-resident Kupffer cells, leading to insulin resistance and nonalcoholic steatohepatitis (NASH) development. However, treatment options that target macrophage/Kupffer cells recruitment or activation to halt the insulin resistance and NASH remain limited. A C-C chemokine receptor 2 (CCR2) plays a central role in macrophage recruitment. In addition, we reported that a different C-C chemokine receptor, CCR5, promotes insulin resistance and hepatic steatosis by regulating macrophage M1/M2 status. Here, we investigated the effect of cenicriviroc (CVC), a dual CCR2 and CCR5 receptor antagonist in a lipotoxic model of NASH with hepatic fibrosis and insulin resistance. Eight-week-old C57BL/6 mice were fed a high-cholesterol, high-fat (CL) diet or a CL diet containing 0.015% CVC (CL+CVC) for 12 weeks. The liver histology, insulin sensitivity, and fibrogenesis were examined. The CVC administration decreased the increase in plasma ALT levels as well as liver triglyceride, cholesterol, and TBARS levels caused by the CL diet. In addition, CVC improved glucose intolerance and hyperinsulinemia in the CL group and augmented the insulin signal, assessed by IRβ and Akt phosphorylation in the liver. Flow cytometry analysis revealed that mice fed CL+CVC had 72% fewer CD11c+CD206- M1-like macrophages, but 94% more CD11c-CD206+M2-like macrophages than CL group, indicating that CVC caused an M2-dominant shift in macrophages/Kupffer cells in the liver. Furthermore, CVC attenuated hepatic fibrosis by repressing the activation of hepatic stellate cells (HSCs) and decreasing hydroxyproline content. Thus, CVC ameliorated lipotoxicity-induced insulin resistance and NASH with hepatic fibrosis by polarizing M2 macrophage and attenuating HSC activation. Disclosure T. Ota: None. G. Chen: None. M. Nagashimada: None. Funding Japan Ministry of Education, Culture, Sports, Science and Technology

Xanthine Oxidase Inhibition Attenuates Insulin Resistance and Diet-Induced Steatohepatitis in Mice

Background: Hyperuricemia drives the development of nonalcoholic fatty liver disease (NAFLD). Pharmacological inhibition of xanthine oxidase (XO), a rate-limiting enzyme for uric acid (UA) production, has been demonstrated to improve hepatic steatosis in diet-induced obese mice. However, it remains unclear whether inhibition of XO could improve nonalcoholic steatohepatitis (NASH), a more advanced form of NAFLD, in terms of both liver inflammation and fibrosis. Methods: To investigate the effects of febuxostat and allopurinol, XO inhibitors clinically used for gout, on diet-induced NASH, C57BL/6J mice were fed a high-fat, high-cholesterol, and cholate diet (CL diet) with or without either 0.001% febuxostat or 0.003% allopurinol for 18 weeks. Systemic glucose tolerance and insulin sensitivity, liver histology, inflammation, and oxidative stress were tested. Furthermore, we conducted a single-arm, open-label intervention study with febuxostat for NAFLD patients with hyperuricemia. Findings: Despite a similar hypouricemic effect of the XO inhibitors on blood UA level, febuxostat, but not allopurinol, significantly decreased hepatic UA levels in CL diet-fed mice. This reduction in hepatic UA levels was accompanied by alleviated insulin resistance, lipid peroxidation, and classically activated M1-like macrophage accumulation in the liver. Furthermore, in NAFLD patients with hyperuricemia, treatment with febuxostat for 24 weeks decreased the serum UA level, accompanied by a reduction in serum levels of liver enzymes, alanine aminotransferase and aspartate aminotransferase. Interpretation: XO may represent a promising therapeutic target in NAFLD/NASH, especially in patients with hyperuricemia. Trial Registration: The study has been registered with UMIN-CTR (#UMI08686). Funding Statement: This work was supported by Grants-in-aid for Scientific Research (B) (25282017) and Challenging Exploratory Research (15K12698) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Japan Diabetes Foundation (to T.O.). This study was also supported by Teijin Pharma Limited. Declaration of Interests: The authors declare no conflict of interest associated with this manuscript. Ethics Approval Statement: The study was approved by the Ethics Committee of Kanazawa University Hospital. All animal procedures were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals of Kanazawa University, Japan.

A Porcine Placental Extract Alleviates Lipotoxicity-Induced NASH by Polarizing M2 Macrophages and Attenuating Stellate Cell Activation in Mice

NAFLD is caused by excessive hepatic lipid accumulation and associated with activation of macrophage/Kupffer cells and oxidative stress, resulting in nonalcoholic steatohepatitis (NASH) with advanced fibrosis. Placental extracts have been used to treat various chronic diseases due to their antioxidative effect. However, the effects of the extracts on the development of NASH have yet to be elucidated. Here, we investigated the effect of porcine placental extract (PPE) in a lipotoxicity-induced NASH model. C57BL/6 mice were fed a high-cholesterol, high-fat (CL) diet or a CL diet containing 0.1% PPE (CL+0.1%PPE) or 0.3% PPE (CL+0.3%PPE). Liver histology and fibrogenesis were examined. In addition, intrahepatic immune cell numbers were quantified by flow cytometry. After 15 weeks of administration, PPE alleviated lipid accumulation by reducing hepatic TG, TC and NEFA levels, as well as lipid peroxidation, which was assessed by TBARS. PPE improved glucose intolerance and enhanced the insulin signal, assessed by IRβ and Akt phosphorylation, in the liver, which was associated with the attenuation of MAPK (ERK/p38MAPK) and NF-kB activation. Flow cytometry analysis revealed that PPE reduced CD11c+CD206−(M1) by 44.2%, whereas it increased CD11c-CD206+(M2) by 1.4-fold, resulting in a predominance of M2 over M1 macrophage populations in the liver of NASH mice. In addition, PPE downregulated LPS-induced M1 marker mRNA expression in isolated murine peritoneal macrophages, but augmented IL-4-induced M2 marker mRNA expression in a dose-dependent manner. Furthermore, the PPE reduced the hepatic stellate cell (HSC) activation associated with the attenuated transforming growth factor-β/Smad3 signaling, both in the liver of NASH mice and in HSC line. Thus, PPE may be a potential approach to prevent NASH by limiting lipid peroxidation, promoting M2 macrophage polarization, and attenuating HSC activation. Disclosure G. Chen: None. L. Xu: None. N. Nagata: None. M. Nagashimada: None. T. Ota: None.

Lactobacillus Pentosus Strain S-PT84 Attenuates Insulin Resistance and Steatohepatitis by Maintaining Gut Permeability and Polarizing M2 Macrophages

Obesity or high-fat diet enhances gut permeability and metabolic endotoxemia, which can trigger insulin resistance and NASH. However, there are few promising treatments targeting lipotoxicity-mediated endotoxemia in NASH. The Lactobacillus pentosus strain S-PT84 has immunomodulatory functions including Th1/Th2 balance modulatory effects and natural killer (NK)/NKT cell activation. In this study, we examined the effect of S-PT84 on diet-induced NASH. C57BL/6 mice were fed a high-cholesterol/high-fat diet (CL) alone or with 1×1010 S-PT84 (CL + S-PT) for 22 weeks. We quantified intrahepatic immune cells using a fluorescence-activated cell sorter (FACS). S-PT84 administration improved hepatic steatosis by decreasing TG and FFA levels by 34% and 37%, respectively. S-PT84 also inhibited the development of hepatic inflammation and fibrosis, lowering F4/80+ macrophage/Kupffer cell infiltration and the hydroxyproline content of the liver. S-PT84 administration in mice fed a CL diet led to improved hyperinsulinemia as well as enhanced hepatic insulin signal assessed by phospho-Akt. FACS analysis revealed that mice fed the CL+S-PT had 71% more CD11c-CD206+ M2 macrophages than mice fed the CL diet, resulting in a predominance of the M2 over the M1 macrophage population. S-PT84, however, showed little effect on NK/NKT cell and T regulatory cell populations in the liver. Furthermore, CL diet increased intestinal permeability as measured with the carboxyfluoresce assay and increased plasma lipopolysaccharide binding protein (LBP) levels. S-PT84 diminished increased intestinal permeability and plasma LBP levels. Additionally, S-PT84 reduced IL-17 producing T cells and increased ZO-1, occludin, and claudin proteins constituting tight junction in colon. In conclusion, S-PT84 treatment attenuated lipotoxicity-induced insulin resistance and NASH by maintaining gut permeability and polarizing M2 liver macrophages. Disclosure Y. Sakai: None. M. Nagashimada: None. T. Ota: None.

Honokiol attenuates diet‐induced non‐alcoholic steatohepatitis by regulating macrophage polarization through activating peroxisome proliferator‐activated receptor γ

Non-alcoholic steatohepatitis (NASH) may develop into hepatic cirrhosis. This study aimed to investigate whether honokiol could prevent NASH induced by high-cholesterol and high-fat (CL) diet in mice and the possible mechanism involved. Mice were fed with CL diet for 12weeks to establish a NASH model; honokiol (0.02% w/w in diet) was added to evaluate its effect on NASH. Murine peritoneal macrophages, RAW264.7 and ANA-1 cells, were used to explore the possible mechanisms of honokiol on macrophage polarization. Mice developed NASH after fed with CL diet for 12weeks. Honokiol supplementation alleviated insulin resistance, hepatic steatosis, inflammation, and fibrosis induced by CL diet. Immunohistochemistry showed that honokiol induced more M2 macrophages in livers compared with CL diet alone. Honokiol decreased M1 marker genes (TNFα and MCP-1) and increased M2 marker gene (YM-1, IL-10, IL-4R and IL-13) expression in mice liver compared with CL diet. Moreover, treatment with honokiol lowered alanine aminotransferase and aspartate aminotransferase in serum and preserved liver from lipid peroxidation, evidenced by lowered hepatic malondialdehyde level. Honokiol has antioxidant function, as honokiol upregulated hepatic glutathione and superoxide dismutase level and downregulated hepatic CYP2E1 protein level. Hepatic peroxisome proliferator-activated receptor γ (PPARγ) and its target genes were upregulated by honokiol. Furthermore, honokiol (10μM) treatment in mouse peritoneal cells, RAW264.7 cells and ANA-1 cells, led to M2 macrophage polarization, whereas a PPARγ antagonist, GW9662, abolished this effect of honokiol. Honokiol can attenuate CL diet-induced NASH and the mechanism in which possibly is polarizing macrophages to M2 phenotype via PPARγ activation.

Effects of Dietary Carbohydrate Sources on Growth and Body Composition of Juvenile Abalone (Haliotis discus, Reeve)

ABSTRACT A 16-wk feeding trial was conducted to assess the effects of different types of dietary carbohydrate on growth and body composition of juvenile abalone (Haliotis discus). A total of 1,680 abalone were randomly distributed among 24 containers (70 per container) and fed one of eight diets, including seven experimental diets containing different types of carbohydrates, including dextrin (DT), glucose (GC), corn starch (CS), α-cellulose (CL), maltose (MT), sucrose (SC), and wheat flour (WF), and Undaria to compare effect of experimental diets. Water stability of the diets was measured 12, 24, and 48 h after seawater immersion. Water stability of the diets was different over all periods of time and their significant interaction was also observed. Survival was not significantly affected by diet. Weight gain and specific growth rate were greatest in abalone fed on the CL diet. Shell length, width, height, and soft body weight were greatest in abalone fed the CL diet, followed by the WF, DT, SC, CS, GC, ...

The Iodine Content in Urine, Faeces and Selected Organs of Rats Fed Lettuce Biofortified with Iodine Through Foliar Application.

Iodine is an essential trace element for humans. Foliar application of micronutrients is successfully used in order to increase the concentration of essential elements in vegetables. The aim of this study was to evaluate the iodine absorption in the rat organism fed foliar biofortified lettuce. The presented study was consisted of the vegetative and animal experiment. In the vegetative experiment with lettuce, two combinations of foliar application were used: (1) control—without iodine application and (2) iodine application in the potassium iodide (KI) form. In the animal experiment, Wistar rats were divided to four groups, which received one of four diets: (1) C—control diet containing iodine in the KI form, (2) D—diet deficient in iodine, (3) D + BL—diet containing biofortified lettuce, and (4) D + CL—diet containing control lettuce (as the only source of iodine in diet, respectively). The diets contained 0.260, 0.060, 0.254 and 0.075 mg I/kg, respectively. In order to determine the iodine absorption in the rat organisms, the content of this trace element was measured in urine, faeces and in selected organs with the use of the ICP-OES technique. Foliar application of the KI increased the content of iodine in lettuce. The rats from the D + BL group excreted significantly less iodine in their urine and faeces and also accumulated more iodine in the organs than the rats from the C group. Iodine with biofortified lettuce was much bioavailable for rodents than iodine from control diet. Biofortified lettuce can be a source of iodine in a diet of human and can improve iodine nutrition.

Plasma Concentrations of GLP-1 and PYY in Rats Fed Dietary Fiber Depend on the Fermentability of Dietary Fiber and Respond to an Altered Diet

Background: It was reported to be associated with intake of dietary fiber and gut hormone. We compared the effect of dietary fiber on the plasma concentration of glucagon-like peptide-1 (GLP-1) and peptide YY (PYY). Methods: Firstly, male Wistar rats (10 wk old) were fed a fiber-free diet (FF diet, AIN-93G-based diet without cellulose) or one of nine FF diets with 5% dietary fiber for 21 days. Cellulose, guar gam (GG), partially hydrolyzed guar gum (PHGG), sugar beet fiber (BF), konjac mannnan (KM), raffinose (RF), hydroxypropyl starch (HPS), hydroxypropyl distarch phosphate (HPDP) and Akamoku (AK) were used as dietary fiber sources. Secondly, we then determined whether the concentrations of GLP-1 and PYY in portal and aorta plasma and microbiota in cecal contents responded to an altered diet. Male Wistar rats (10 wk old) were fed the PHGG diet for 20 days before being changed to the FF diet. Results: The amount of short chain fatty acids (SCFA) in the cecal contents markedly increased in rats fed the GG, PHGG, BF, KM, RF, HPS or HPDP diets compared with SCFA in rats fed the FF, CL or AK diets. The concentrations of GLP-1 and PYY in portal plasma positively correlated with the concentration of SCFA in the cecal contents and cecal tissue weight. The concentrations of GLP-1 and PYY, in the altered diet group decreased to the same concentration as that measured in the FF group. However, it was low the similarities in the cecal bacteria population. Conclusion: The plasma concentrations of GLP-1 and PYY increased with increasing the amounts of SCFA, the products of DF fermentation, in the cecal contents. GLP-1 and PYY plasma concentrations and SCFA concentrations in the cecal contents rapidly responded to the altered diet, but the cecal bacteria population did not.

Suppression of Mammary Carcinogenesis Through Early Exposure to Dietary Lipotropes Occurs Primarily In Utero

The study determined whether feeding during lactation affects the suppressive effect of maternal dietary lipotropes (i.e., methionine, choline, folate, and vitamin B12) on mammary carcinogenesis. Pregnant Sprague-Dawley rats were randomly allocated to the control diet during pregnancy and lactation (CC), lipotropes-fortified diet during pregnancy (LC), lipotropes-fortified diet during pregnancy plus lactation (LL), or lipotropes-fortified diet during lactation (CL). Randomly selected female offspring from each group were injected intraperitoneally with 50 mg/kg body weight of N-nitroso-N-methylurea at 50 days of age to induce mammary tumors. The LC and LL diets significantly increased tumor latency and survival (P < 0.05). Tumor volumes were significantly suppressed in LC and LL offspring as compared with the CC and CL pups (3759.1 ± 563.0 and 3603.7 ± 526.1 vs. 7465.0 ± 941.1 and 5219.3 ± 759.8 mm3, respectively; P < 0.05). Both LC and LL lowered tumor multiplicity as compared with CC and CL (P < 0.05). The LC and LL diets repressed transcription of histone deacetylase (HDAC) 1 as well as total HDAC enzyme activity as compared with CC and CL diets (P < 0.05). Data suggest that the tumor suppressive effect of maternal dietary lipotropes is primarily in utero and may be linked to regulation of proteins involved in chromatin remodeling.

Prevention and reversal of lipotoxicity-induced hepatic insulin resistance and steatohepatitis in mice by an antioxidant carotenoid, β-cryptoxanthin.

Excessive hepatic lipid accumulation promotes macrophages/Kupffer cells activation, resulting in exacerbation of insulin resistance and progression of nonalcoholic steatohepatitis (NASH). However, few promising treatment modalities target lipotoxicity-mediated hepatic activation/polarization of macrophages for NASH. Recent epidemiological surveys showed that serum β-cryptoxanthin, an antioxidant carotenoid, was inversely associated with the risks of insulin resistance and liver dysfunction. In the present study, we first showed that β-cryptoxanthin administration ameliorated hepatic steatosis in high-fat diet-induced obese mice. Next, we investigated the preventative and therapeutic effects of β-cryptoxanthin using a lipotoxic model of NASH: mice fed a high-cholesterol and high-fat (CL) diet. After 12 weeks of CL diet feeding, β-cryptoxanthin administration attenuated insulin resistance and excessive hepatic lipid accumulation and peroxidation, with increases in M1-type macrophages/Kupffer cells and activated stellate cells, and fibrosis in CL diet-induced NASH. Comprehensive gene expression analysis showed that β-cryptoxanthin down-regulated macrophage activation signal-related genes significantly without affecting most lipid metabolism-related genes in the liver. Importantly, flow cytometry analysis revealed that, on a CL diet, β-cryptoxanthin caused a predominance of M2 over M1 macrophage populations, in addition to reducing total hepatic macrophage and T-cell contents. In parallel, β-cryptoxanthin decreased lipopolysaccharide-induced M1 marker mRNA expression in peritoneal macrophages, whereas it augmented IL-4-induced M2 marker mRNA expression, in a dose-dependent manner. Moreover, β-cryptoxanthin reversed steatosis, inflammation, and fibrosis progression in preexisting NASH in mice. In conclusion, β-cryptoxanthin prevents and reverses insulin resistance and steatohepatitis, at least in part, through an M2-dominant shift in macrophages/Kupffer cells in a lipotoxic model of NASH.