Involvement of the vagus nerve and hepatic gene expression in serum adiponectin concentrations in mice.

Several humoral factors, such as adiponectin and urate, have been suggested to affect metabolic syndromes. Previously, we reported a reduction in blood adiponectin concentrations after a high-fructose diet partially via the vagus nerve in rats. Although a lithogenic diet (LD), i.e., supplementation of a normal control diet (CT) with 0.6% cholesterol and 0.2% sodium cholate, reduced blood adiponectin concentrations, the involvement of the vagus nerve in this mechanism remains unclear. To estimate the involvement of the vagus nerve in the regulation of blood adiponectin concentrations using an LD, male imprinting control region mice that had been vagotomized (HVx) or only laparotomized (Sham) were administered a CT or an LD for 10 weeks. Serum adiponectin concentrations in the Sham-LD, HVx-CT, and HVx-LD groups were reduced by half compared with the Sham-CT group. The hepatic mRNA levels of fibroblast growth factor 21 (Fgf21), which reportedly stimulates adiponectin secretion from white adipose tissue, were lower in the LD groups compared with the CT groups. HepG2 hepatoma cells showed that various bile acids reduced the mRNA expression of FGF21. Moreover, the LD increased serum urate concentrations and reduced hepatic expressions of the acyl-CoA oxidase 1 (Acox1) mRNA and glucokinase, suggesting insufficient regeneration of ATP from AMP. In conclusion, serum adiponectin concentration may be regulated via the vagus nerve in normal mice, whereas a reduction of hepatic Fgf21 mRNA by bile acids may also lower serum adiponectin levels. Moreover, the LD may promote hepatic AMP accumulation and subsequently increase the serum urate concentration in mice.

Beneficial health effects of polyphenols metabolized by fermentation.

High daily intake of polyphenol-rich meal in some countries could be regarded as a healthy meal. However, the knowledge about the bioavailability and functionality of the exiting amounts of polyphenol into the large intestine needs to be elucidated, particularly the beneficial health effects and its fermentation characteristics during fermentation. Thus, this review focuses on the influence of polyphenols metabolized by fermentation and elucidates their health attributes. Besides, it also summarized the potential benefits of polyphenols and discussed the need for further research to fully understand the health attributes of polyphenols.

Dietary Ethanolamine Plasmalogen Alleviates DSS-Induced Colitis by Enhancing Colon Mucosa Integrity, Antioxidative Stress, and Anti-inflammatory Responses via Increased Ethanolamine Plasmalogen Molecular Species: Protective Role of Vinyl Ether Linkages.

Dietary ethanolamine plasmalogen (PlsEtn) has been reported to have several health benefits; however, its functional role during colon pathophysiology remains elusive. The present study investigated the anticolitis effect of dietary ethanolamine glycerophospholipids (EtnGpls) with high PlsEtn from ascidian muscle (86.2 mol %) and low PlsEtn from porcine liver (7.7 mol %) in dextran sulfate sodium (DSS)-induced colitis in mice. Dietary EtnGpls lowered myeloperoxidase activity, thiobarbituric acid-reactive substances, proinflammatory cytokines and proapoptosis-related protein levels in colon mucosa after 16 days of DSS treatment, with ascidian muscle (0.1% EtnGpl in diet) showing higher suppression than porcine liver (0.1% EtnGpl in diet). Moreover, dietary EtnGpls suppressed DSS symptoms after 38 days of DSS treatment as evidenced by increased body weight, colon length, and ameliorated colon mucosa integrity. Additionally, dietary EtnGpls elevated short-chain fatty acid production in DSS-treated mice. Altogether, these results indicate the potential of utilizing diets with abundant PlsEtn for the prevention of colon inflammation-related disorders.

Combined effects of BARLEYmax and cocoa polyphenols on colonic microbiota and bacterial metabolites in vitro.

BARLEYmax, a barley variety, and cocoa polyphenols (CPPs) have been reported to affect bacterial metabolites in the colon. This study aimed to evaluate the combined effects of BARLEYmax and CPPs supplementation on fecal microbiota in vitro using pig feces for 48 h. The relative abundances of the family Clostridiaceae and the genus Clostridium and ammonia-nitrogen production were decreased by both BARLEYmax and CPP supplementation, and there was a positive correlation between their abundances and the ammonia-nitrogen concentration. Although acetate and n-butyrate production was decreased by CPP supplementation, their concentrations were maintained at a higher level in the BARLEYmax + CPP group than in the cellulose (control) and cellulose + CPP groups. Therefore, this study demonstrated that a combination of BARLEYmax and CPPs may be beneficial in maintaining higher short-chain fatty acid production and the elimination of potentially harmful factors. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00959-z.

Bile acids induced hepatic lipid accumulation in mice by inhibiting mRNA expression of patatin-like phospholipase domain containing 3 and microsomal triglyceride transfer protein.

Preliminary studies have shown that a lithogenic diet (LG), which contains cholesterol and cholic acid, induces gallstones and hepatic lipid accumulation (HLA), and reduction of blood triglyceride in mice. We hypothesized that an LG induces HLA by diminishing hepatic triglyceride excretion; however, there is no clear understanding of the mechanism of LG-induced HLA. This study aimed to investigate transcript expression related to the synthesis, expenditure, and efflux of hepatic triglyceride, in mice fed an LG for 4 weeks. Results showed lower plasma concentrations of triglyceride in the LG group than in the control group, but no symptoms of hepatic injury were observed. Hepatic mRNA expressions of patatin-like phospholipase domain containing 3 (Pnpla3), microsomal triglyceride transfer protein (Mttp), and acyl-CoA oxidase 1 (Acox1) were also reduced in the LG group. Deoxycholic acid and lithocholic acid promoted intracellular lipid accumulation, reduced triglyceride concentration in media, and suppressed expression of PNPLA3 and MTTP in HepG2 human hepatoma cells. These findings suggest that deoxycholic acid and lithocholic acid promote HLA by inhibiting the expression of PNPLA3, ACOX1, and MTTP that are involved in lipid metabolism.

Effect of polyphenols isolated from purple sweet potato (Ipomoea batatas cv. Ayamurasaki) on the microbiota and the biomarker of colonic fermentation in rats fed with cellulose or inulin.

A polyphenol-rich diet has been associated with various health benefits. This study assessed the effects of polyphenol/anthocyanin isolated from a purple sweet potato (Ipomoea batatas cv. Ayamurasaki) on colonic fermentation in cellulose- or inulin-fed rats. Male Fischer-344 rats were assigned to one of these experimental diets: 5% cellulose (CEL), 5% CEL + 1% purple sweet potato polyphenol extract (CELP), 5% inulin (INU), and 5% INU + 1% purple sweet potato polyphenol extract (INUP) in each diet. The purple sweet potato polyphenol extract (PSPP) increased the relative abundance of Dorea and reduced the relative abundances of Oscillospira and Bacteroides in cellulose- or inulin-fed rats, respectively. Besides, PSPP reduced the caecal iso-butyrate and pH in the cellulose-fed rats. Further, PSPP triggered an increase in the caecal mucin level when combined with cellulose and increased the caecal IgA level while reducing the indole production in both the cellulose- or inulin-fed rats. Finally, PSPP may have different effects on the intestinal fermentation properties depending on the fermentability of dietary fiber associated with it. Therefore, this study demonstrated that dietary inclusion of polyphenol/anthocyanin from purple sweet potato might confer positive health attributes to the host gut.

Purple Sweet Potato Polyphenols Differentially Influence the Microbial Composition Depending on the Fermentability of Dietary Fiber in a Mixed Culture of Swine Fecal Bacteria.

The prevalence of many chronic diseases which have been associated with poor nutrition may be reduced by the positive modulation of colonic microbiota. In this study, we assess the effects of purple sweet potato polyphenols (PSP) in a mixed culture of swine fecal bacteria during in vitro colonic fermentation using pig colonic digest. Jar fermenters were used to conduct a small scale in vitro colonic fermentation experiments under the anaerobic condition for 48 h. Jar fermenters were assigned to one of the following groups: Cellulose, cellulose + PSP, inulin, and inulin + PSP. The present study revealed that the polyphenolic content of purple sweet potato could modulate the colonic microbiota by differentially increasing the population of beneficial bacteria and decreasing the pathogenic bacteria depending on cellulose and inulin. Accordingly, PSP might be a material conducive for improving the conditions for the fermentation of partly-fermentable dietary fiber. Besides, PSP was also responsible for the drastic reduction of putrefactive products, especially p-cresol to a significant level. Our results suggest that PSP could alter the microbial composition depending upon the fermentability of dietary fiber and has the potential to maintain a stable and healthy colonic environment that will ultimately alleviate chronic diseases development and confer health benefits to the host.

Ultrastructural changes in colonic epithelial cells in a rat model of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a global, chronic intractable disease. The functions of drugs and food components have been evaluated in models of IBD induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Here, we used transmission (TEM) and osmium-maceration scanning (SEM) electron microscopy to evaluate the ultrastructure of colonic epithelial cells in rat models of IBD induced by TNBS. Histological evaluation revealed that the intestinal crypts in the most regions of the IBD-model colons were deformed and we classified them as having high cell migration rates (HMIG). The remaining regions in the intestinal crypts retained a relatively normal structure and we classified them as having low cell migration rates (LMIG). Osmium-maceration SEM revealed the mucosal fluid flowing in spaces without secretory granules in crypt goblet cells of both HMIG and LMIG regions, indicating the depletion of goblet cell mucin that is found in patients with IBD. The Golgi apparatus in absorptive cells was stacked and curled in both regions. Osmium-maceration SEM showed membrane network structures resembling endoplasmic reticulum that were large and expanded in absorptive cells with HMIG rather than with LMIG regions in IBD-model colons. These findings indicated that endoplasmic reticulum stress is associated with susceptibility to IBD and that the effects of various agents can be evaluated according to endoplasmic reticulum stress revealed by using electron microscopy in models of IBD induced by TNBS.

Safety and efficacy of adzuki bean extract in subjects with moderate to high LDL-C: a randomized trial.

The safety and efficacy of polyphenol-containing adzuki bean extract on lipid metabolism were evaluated in human subjects in an 8-week, randomized, double-blind, placebo-controlled, parallel intervention study. No adverse effects were observed in the participants receiving adzuki bean extract. The adzuki bean group showed a significant increase in the ΔHDL-C concentration compared with the placebo group after 4 weeks of intervention (3.76 ± 7.79 mg/dL vs. -0.08 ± 6.03 mg/dL), respectively, and both groups showed reduced ∆HDL-C concentrations, with the adzuki bean extract group showing a return to the baseline levels (0.36 ± 5.36 mg/dL) and the placebo group showing a decrease to below the baseline levels (-3.17 ± 7.79 mg/dL) at week 8. This short-term study represents the first step in establishing the practicality, safety, and plausibility of HDL-C maintaining effects of adzuki bean extract in human subjects.

Effect of a combination of inulin and polyphenol-containing adzuki bean extract on intestinal fermentation in vitro and in vivo.

The effect of a combination of inulin (INU) and polyphenol-containing adzuki bean extract (AE) on intestinal fermentation was examined in vitro using fermenters for 48 h and in vivo using rats for 28 d. The total short-chain fatty acid concentrations in the fermenters were decreased by a combination of INU and AE, but the concentration in the INU + AE group was higher than the cellulose (CEL) and CEL + AE groups. The cecal propionate concentration was increased by a combination of INU and AE compared with their single supplement. The ammonia-nitrogen concentration in the fermenters and rat cecum was decreased by INU and AE. Cecal mucin levels were increased by INU and AE respectively. Therefore, our observations suggested that the combination of INU and AE might be a material of functional food that includes several healthy effects through intestinal fermentation.

Intraduodenal infusion of cyanidin-3-glucoside transiently promotes triglyceride excretion into bile in rats.

Flavonoids purportedly have a role in improving lipid metabolism. In our preliminary study, highly concentrated flavonoid metabolites appeared in bile juice in rats, which also contains various lipids. Biliary flavonoid metabolites generally have amphiphilic properties, may influence lipid solubility, and possibly contribute to the improvement of dyslipidemia. However, the influence of biliary flavonoid metabolites on the biliary lipid profile is not well known. Therefore, we hypothesized that the amphiphilic property of biliary flavonoid metabolites alters biliary lipid profiles. To estimate the influence of flavonoids on the biliary lipid profile, we laparotomized rats under anesthesia, intraduodenally injected them with cyanidin-3-glucoside chloride (C3G) or quercetin, and analyzed their biliary metabolite concentrations for 2 hours. Concentrations of C3G and quercetin metabolites peaked at 30 minutes after the injection; those of quercetin were 6 to 10 times higher than those of C3G throughout the sampling period up to 2 hours. Biliary triglyceride (TG) concentrations were higher in the C3G group at 30 and 45 minutes; biliary cholesterol and phospholipid concentrations were lower in the quercetin group at 30 minutes than those in the control group. Hepatic TG content after the 2-hour sampling was lower in the C3G group than in the control group. These results suggest that C3G, but not quercetin, may transiently promote TG excretion into bile, with a reduction in hepatic TG content. This C3G effect may be involved in improvement of TG metabolism.

Relationships among alcoholic liver disease, antioxidants, and antioxidant enzymes.

Excessive consumption of alcoholic beverages is a serious cause of liver disease worldwide. The metabolism of ethanol generates reactive oxygen species, which play a significant role in the deterioration of alcoholic liver disease (ALD). Antioxidant phytochemicals, such as polyphenols, regulate the expression of ALD-associated proteins and peptides, namely, catalase, superoxide dismutase, glutathione, glutathione peroxidase, and glutathione reductase. These plant antioxidants have electrophilic activity and may induce antioxidant enzymes via the Kelch-like ECH-associated protein 1-NF-E2-related factor-2 pathway and antioxidant responsive elements. Furthermore, these antioxidants are reported to alleviate cell injury caused by oxidants or inflammatory cytokines. These phenomena are likely induced via the regulation of mitogen-activating protein kinase (MAPK) pathways by plant antioxidants, similar to preconditioning in ischemia-reperfusion models. Although the relationship between plant antioxidants and ALD has not been adequately investigated, plant antioxidants may be preventive for ALD because of their electrophilic and regulatory activities in the MAPK pathway.

Porcine Splenic Hydrolysate has Antioxidant Activity in vivo and in vitro.

The antioxidant capacity of porcine splenic hydrolysate (PSH) was studied in vitro and in vivo. Peptide hydrolysates were prepared, using the proteolytic enzyme Alcalase(®). The molecular weights of PSH were 37,666, 10,673, 6,029, and 2,918 g/mol. Rats were fed a 5% (w/v) PSH diet, instead of a casein diet, for 4 wk. The food intake, body weight gain, and liver weight of rats in the PSH group were similar to those in the control (CONT) group. There were no differences in the serum total cholesterol, triglyceride, total protein, or albumin levels between PSH and CONT groups. However, the level of in vivo hepatic lipid peroxidation in PSH group was significantly lower than that in CONT. In vivo hepatic catalase and glutathione peroxidase activities in the PSH group were significantly higher than those in the control group. The in vitro protein digestibility of PSH was lower than that of casein. The in vitro trolox equivalent antioxidant capacity of PSH was significantly higher than that of the peptide hydrolysate from casein. The in vitro radical scavenging activities of PSH were significantly higher than those of the peptide hydrolysate from casein. The present findings suggest that porcine splenic peptides improve the antioxidant status in rats by enhancing hepatic catalase and GSH-Px activities, and indicate a potential mechanism of radical scavenging activity during gastrointestinal passage.

Dietary L-cysteine improves the antioxidative potential and lipid metabolism in rats fed a normal diet.

L-cysteine works as a precursor of the antioxidant, glutathione. We investigated the effects of L-cysteine (1% and 2%) on lipid metabolism and the antioxidative system in rats fed a normal diet. Administering L-cysteine dependently decreased the food intake, fat mass weight and body weight dose. Dietary L-cysteine also decreased the triglyceride levels in the serum and liver. However, there were no significant differences in the hepatic TBARS and glutathione (GSH) levels among the groups. The activities of catalase and glutathione reductase in the rats receiving 2% L-cysteine were significantly higher (p<0.05) than in the control rats. These results suggest that dietary L-cysteine dose-dependently affected the antioxidative enzyme activities, and the lipid levels in the serum and liver which might be related to the reduced food intake.

Japanese butterbur (Petasites japonicus) leaves increase hepatic oxidative stress in male rats.

We investigated the adverse effects of Japanese butterbur leaves (Petasites japonicus, Compositae) in male F344/DuCrj rats. The rats were fed a control diet or a treatment diet containing 5% butterbur leaf powder for 4 weeks. No differences were observed in body weight gain, food intake or feed efficiency between treatments, but relative liver weight in the butterbur group was significantly higher than that of the control group. In addition, thiobarbituric acid reactive substances (TBARs) and glutathione levels in the serum and liver of the butterbur group were higher than those of the control group. Hepatic glutathione reductase and glutathione S-transferase activities and mRNA expression in the butterbur leaf group were higher than in the control group. Furthermore, hepatic cytochrome 2E1 mRNA expression was higher than in the control group. In vitro, an acetone extract of the butterbur leaf powder showed the strongest increase in TBARs level in a hepatic homogenate through 4 d. Our findings suggest that feeding 5% butterbur leaf powder to rats can cause adverse effects by increasing oxidative stress.

Anti-obesity role of adzuki bean extract containing polyphenols: in vivo and in vitro effects.

BACKGROUND: The aim of this work was to evaluate the effects of polyphenol-rich adzuki bean extract on lipid metabolism, triglyceride accumulation and proinflammatory cytokine secretion in vivo and in vitro. RESULTS: For the in vivo study, rats were divided into four groups: group C was fed a control diet, group A was fed the control diet with 1% adzuki bean extract, group CF was fed a high fat diet, and group AF was fed a high fat diet with 1% adzuki bean extract. For the in vitro study, the ability of adzuki bean extract to suppress triglyceride incorporation, glycerol phosphate dehydrogenase activity and inflammatory response was investigated in cultured human adipocytes. Data from the animal study showed that adzuki bean extract improved lipid metabolism in both the normal and high-fat diet groups. Adzuki bean extract treatment in the high-fat group resulted in significant reductions in total hepatic lipid accumulation and lipid secretion into the feces. Incubation of adipocytes with adzuki bean extract significantly decreased triglyceride accumulation, glycerol phosphate dehydrogenase activity and inflammatory responses without affecting cell viability. CONCLUSION: The results of this study demonstrate that adzuki bean extract has high potential to serve as a natural anti-obesity agent.

Effects of dietary supplementation with betaine on a nonalcoholic steatohepatitis (NASH) mouse model.

The effects of betaine supplementation on non-alcoholic steatohepatitis (NASH) model mice were examined by measuring the accumulation of fat in the livers of NASH model mice compared to a control. Betaine from sugar beets was provided to the model mice as a dietary supplement. After 3 wk of dietary supplementation, there were no significant differences in body weight or liver weight between the groups. However, the liver to body weight ratio in the high-fat diet with betaine (HFB) group was significantly (p<0.05) higher than that in the high-fat diet (HF) group. There were no differences in serum triglyceride (TG) concentrations, AST and ALT activities, or hepatic glutathione concentrations between the groups. Hepatic TG level in the HFB group was significantly (p<0.05) lower than that in the HF group. Hepatic cells obtained from the HF group showed increased occurrence of explosive puff and necrosis as compared with those in the HFB group. Betaine supplementation had an inhibitory effect on fat accumulation in the liver: the Oil red-positive area in the HFB group (0.82 ± 0.85%) was significantly (p<0.001) smaller than that in the HF group (9.06 ± 2.24%). These results indicate the potential of betaine to serve as an agent for amelioration of hepatic steatosis in NASH model mice.

Amelioration of D-galactosamine-induced acute liver injury in rats by dietary supplementation with betaine derived from sugar beet molasses.

The effects of betaine supplementation on D-galactosamine-induced liver injury were examined in terms of hepatic and serum enzyme activities and of the levels of glutathione and betaine-derived intermediates. The rats induced with liver injury showed marked increases in serum enzyme activity, but those receiving dietary supplementation of 1% betaine showed enzyme activity levels similar to a control group without liver injury. Administration of betaine also increased both hepatic and serum glutathione levels, even following D-galactosamine injection. The activity of glutathione-related enzymes was markedly decreased following injection of D-galactosamine, but remained comparable to that of the control group in rats receiving 1% betaine. The concentrations of hepatic S-adenosyl methionine and cysteine showed similar trends to that observed for hepatic glutathione levels. These results indicate that 1% betaine has a hepatoprotective effect by increasing hepatic and serum glutathione levels along with glutathione-related enzyme activities in rats.

Utilization of adzuki bean extract as a natural antioxidant in cured and uncured cooked pork sausages.

A commercial adzuki bean extract (AE) was evaluated for antioxidant effectiveness in cured and uncured cooked pork sausages. TBARS values, instrumental color evaluation and sensory panel scores were assessed. For uncured sausages, AE at 0.2% was equally effective as 0.1% butylated hydroxytoluene (BHT) in reducing TBARS values. Similarly, AE at 0.2% significantly (P<0.01) reduced the TBARS in cured sausages. Incorporation of 0.2% AE into sausages produced higher (P<0.05) CIE lab color a* value and lower (P<0.05) L* and b* values. Sensory panels did not detect any difference in color, odor, taste, flavor, and overall acceptance in uncured pork sausages with addition of 0.2% AE. However, there were adverse changes in the color and odor of cured sausages, even though the taste, flavor, and overall acceptance were similar. Therefore, the results suggest that AE is a potential antioxidant.

Colored potato extracts induce superoxide dismutase-2 mRNA via ERK1/2 pathway in HepG2 cells.

Rats fed a diet containing Shadow Queen (SQ), an anthocyanin-rich potato cultivar, previously showed an increase in the hepatic superoxide dismutase (SOD)-2 mRNA level. We investigated whether an extract of SQ would directly increase the hepatic SOD-2 mRNA level in HepG2 cells. Furthermore, we estimated the intracellular signaling pathway for the induction of SOD-2 mRNA expression. HepG2 cells were stimulated using extracts of four crops, including SQ, for 12 h; only extracts of colored potatoes induced SOD-2 mRNA expression significantly. This induction of SOD-2 mRNA expression was blocked by an inhibitor of the extracellular signal-related kinase (ERK) 1/2 pathway. Furthermore, an extract of SQ increased the phosphorylation of ERK1/2 after 15 or 30 min of stimulation. These data indicate that an extract of SQ directly induces hepatic SOD-2 mRNA expression via activation of ERK1/2 pathway in HepG2 cells.