Butyrate Protects Mice Against Methionine-Choline-Deficient Diet-Induced Non-alcoholic Steatohepatitis by Improving Gut Barrier Function, Attenuating Inflammation and Reducing Endotoxin Levels.

Butyrate exerts protective effects against non-alcoholic steatohepatitis (NASH), but the underlying mechanisms are unclear. We aimed to investigate the role of butyrate-induced gut microbiota and metabolism in NASH development. Sixty-five C57BL/6J mice were divided into four groups (n = 15-17 per group) and were fed either a methionine-choline-sufficient (MCS) diet or methionine-choline-deficient (MCD) diet with or without sodium butyrate (SoB; 0.6 g/kg body weight) supplementation for 6 weeks. Liver injury, systematic inflammation, and gut barrier function were determined. Fecal microbiome and metabolome were analyzed using 16S rRNA deep sequencing and gas chromatography-mass spectrometry (GC-MS). The results showed that butyrate alleviated the MCD diet-induced microbiome dysbiosis, as evidenced by a significantly clustered configuration separate from that of the MCD group and by the depletion of Bilophila and Rikenellaceae and enrichment of promising probiotic genera Akkermansia, Roseburia, Coprococcus, Coprobacillus, Delftia, Sutterella, and Coriobacteriaceae genera. The fecal metabolomic profile was also substantially improved by butyrate; several butyrate-responsive metabolites involved in lipid metabolism and other pathways, such as stearic acid, behenic acid, oleic acid, linoleic acid, squalene, and arachidonic acid, were identified. Correlation analysis of the interaction matrix indicated that the modified gut microbiota and fecal metabolites induced by butyrate were strongly correlated with the alleviation of hepatic injury, fibrosis progression, inflammation, and lipid metabolism and intestinal barrier dysfunction. In conclusion, our results demonstrated that butyrate exerts protective effects against NASH development, and these effects may be driven by the protective gut microbiome and metabolome induced by butyrate. This study thus provides new insights into NASH prevention.

Antioxidant capacity and identification of the constituents of ethyl acetate fraction from Rhus verniciflua Stokes by HPLC-MS.

Ethyl acetate fraction (EAF) from Rhus verniciflua Stokes is an important source of bioactive compounds. The aim of this study was the tentative identification and quantification of phenolic compounds, comparison of the phenolic structure-antioxidant activity relationships. Twelve compounds of EAF belonging to polyphenol types were detected by high performance liquid chromatography and analysed on line with negative ion electrospray ionisation tandem mass spectrometry, which were ethoxy 3-hydroxy benzoic acid, gallic acid (GA), 3,4-dihydroxy amygdalic acid, gallic acid cetyl ester, protocatechuic acid (PA), fustin, ethyl gallate (EG), garbanzol, fisetin, sulfuretin, butin and 3,7-dihydroxyflavanone-4'-rhamnoside. The antioxidant activity were evaluated based on the different types of radical scavenging capacities, i.e. DPPH·, ABTS·+ and OH. The antioxidant capacity of EAF mainly depended on the GA, EG, PA, fisetin, sulfuretin and butin. The phenolics exhibited a dose-dependent behaviour and high antioxidant ability.

In Vivo and In Vitro Toxicity Evaluation of Polyprenols Extracted from Ginkgo biloba L. Leaves.

Polyprenols of Ginkgo biloba L. leaves (GBP) are a new type of lipid with 14-24 isoprenyl units, which in humans have strong bioactivity like the dolichols. A large amount of work showed that GBP had good antibacterial activity and powerful protective effects against acute hepatic injury induced by carbon tetrachloride and alcohol, as well as antitumor activity, but the safety of GBP was not considered. The current study was designed to evaluate the toxicity of these polyprenols. Acute toxicity in mice was observed for 14 days after GBP oral dosing with 5, 7.5, 10, 15 and 21.5 g/kg body weight (b. wt.) Further, an Ames toxicity assessment was carried out by plate incorporation assay on spontaneous revertant colonies of TA97, TA98, TA100 and TA102, with GBP doses designed as 8, 40, 200, 1000 and 5000 µg/dish, and subchronic toxicity was evaluated in rats for 91 days at GBP doses of 500, 1000 and 2000 mg/kg b. wt./day. The weight, food intake, hematological and biochemical indexes, the ratio of viscera/body weight, and histopathological examinations of tissue slices of organs were all investigated. The results showed that no animal behavior and appearance changes and mortality were seen during the observation period with 21.5 g/kg GBP dose in the acute toxicity test. Also, no mutagenicity effects were produced by GBP (mutation rate < 2) on the four standard Salmonella strains (p > 0.05) in the Ames toxicity test. Furthermore, the no observed adverse effect level (NOAEL) of GBP was 2000 mg/kg for 91 days feeding of rats in the subchronic toxicity tests. Results also showed the hematological and biochemical indexes as well as histopathological examination changed within a small range, and all clinical observation indexes were normal. No other distinct impacts on cumulative growth of body weight, food intake and food utilization rate were discovered with GBP. No significant difference was discovered for the rats' organ weight and the ratio of viscera to body weight (p > 0.05). Reversible pathological changes in the histopathological examinations of tissue slices of organs were not observed. GBP could therefore be considered as a safe material with minor side effects.

Effects of dietary supplementation of fermented Ginkgo biloba L. residues on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets.

This study evaluated the effects of fermented Ginkgo biloba L. residues (FGBLR) on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets. Pigs were allotted to five dietary treatments, including negative control (NC: antibiotic free basal diet), positive control (PC) (NC + 30 mg apramycin/kg) and FGBLR-50, 100, 150 (NC + 50, 100, 150 g FGBLR/kg). Pigs in FGBLR-100 and PC treatments showed increased final body weight, average daily gain, gain:feed and apparent total tract digestibility of dry matter, N and gross energy (P < 0.05) compared with NC, FGBLR-50 and FGBLR-150 treatments, In addition, pigs fed with FGBLR-100 diet showed higher serum total protein, albumin, alkaline phosphatase, glucose, hemoglobin, total iron, total iron binding capacity, superoxide dismutase and glutathione superoxide dismutase levels, and lower serum blood urea nitrogen, malondialdehyde, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, triglyceride and total cholesterol levels than those fed with PC and NC diets (P < 0.05). Moreover, feeding FGBLR-100 could increase levels of immunoglobulin G (IgG), IgA and IgM, as well as lymphocyte transformation rates, ratio of CD4+ to CD8+ cells and proportions of CD2+, CD4+, B, major histocompatibility complex (MHC)-I and MHC-II cells, and can decrease proportion of CD8+ cells in blood of piglets compared with PC and NC groups (P < 0.05). These results indicate that dietary supplementation with 10% of FGBLR showed greatest beneficial effects on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets, which were superior to antibiotic supplemental diets.

Chemical constituents and structural characterization of polysaccharides from four typical bamboo species leaves.

In order to find bamboo leaves with high contents of bioactive polysaccharides, 32 samples were chosen to analyze their polysaccharide content by GC and sulfuric acid-anthrone colorimetric assays. Purified polysaccharides (BLPS) were separated from the four varieties P. nigra (Lodd.) Munro (PN), P. vivax McClure (PV), Chimonobambusa quadrangularis (Fenzi) Makino (CQ), and P. bambussoides cv. Tanakae (PB) by ultrasound extraction, solution precipitation, ion exchange resin, DEAE-52 and Sephadex G-100 chromatography. BLPS structural characterization was accomplished by HPLC-GPC, Fourier transform infra-red spectroscopy (FTIR) and NaIO4-HIO4 oxidation reactions. The results showed that the total polysaccharides of the bamboo leaves in samples 1-32 ranged between 1.4% and 5.4%, Samples No. 29-No. 32 (PN, PV, CQ, and PB) contained 2-3 fold more polysaccharides than No. 1~No. 28 among the 32 different species, particularly the content of galactose was in a range of 21.5%-34.1% for these four typical bamboo species leaves, which was also more than 2-3 fold higher than in No. 1-No. 28. Sugar analysis indicated that PN-PBLPS-1, PV-PBLPS-1, CQ-PBLPS-1 and PB-PBLPS-1 from the four varieties were homogeneous polysaccharides with molecular weights of 2.04 × 104, 1.15 × 104, 8.75 × 104 and 1.48 × 104 Da, respectively. PB-PBLPS-1 was a mixture of α-galactopyranose and ß-d-glucopyranose linkages with α-(1→6) or ß-(1→6)glycosidic bonds, while PN-PBLPS-1, PV-PBLPS-1, and CQ-PBLPS-1 had α galactopyranose linkages with α-(1→6) glycosidic bonds.

Antiviral activity of a nanoemulsion of polyprenols from ginkgo leaves against influenza A H3N2 and hepatitis B virus in vitro.

UNLABELLED: In order to improve the bioavailability levels of polyprenols (derived from ginkgo leaves (GBP)) in the human body, a GBP nanoemulsion was prepared, and its antiviral activity was evaluated against influenza A H3N2 and hepatitis B virus in vitro. METHODS: A GBP nanoemulsion was prepared by inversed-phase emulsification (IPE). Next, we investigated the antiviral activity of the GBP nanoemulsion on influenza A H3N2 and hepatitis B virus in vitro by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenlytetrezolium bromide) method. ELISA and the fluorescent quantitative PCR method were used to measure the content of HBsAg, HBeAg and DNA virus in human samples. RESULTS: The GBP nanoemulsion exhibited uniformity at an average particle size 97 nm with a hydrophilic-lipophilic balance (HLB) of 9.5. GBP is non-toxic to normal cells, hepatitis B virus DNA, hepatitis B virus antigen and HepG2215. Furthermore, GBP could reach a 70% virucidal activity and a 74.9% protection rate (*** p < 0.001) on MDCK cells infected with H3N2 virus at a high concentration of 100 µg/mL. GBP had a good inhibition rate on HBsAg (52.11%, ** p < 0.01) at 50 µg/mL and Day 9 of incubation, and a 67.32% inhibition effect on HBeAg at a high concentration of 100 µg/mL and Day 9. GBP had good inhibition on HBV DNA with CT 18.6 and lower copies (** p < 0.01) at a middle concentration of 12.5 to 25 µg/mL. CONCLUSIONS: The GBP nanoemulsion was very stable and non-toxic and had very strong antiviral activity against influenza A H3N2 and hepatitis B virus in vitro. The inhibitory effects and reactive mechanisms were similar to the drug, 3TC; by lengthening the incubation time and increasing the drug concentration, GBP has promising potential as an antiviral drug.

Enzymatic hydrolysis of oleuropein from Olea europea (olive) leaf extract and antioxidant activities.

Oleuropein (OE), the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT) and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity) optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE) were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 µg/mL) was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities.

Decompressing inner ebullition (DIE) can reduce the extraction liquid boiling point and polyphenols oxidation in the extraction process. The aim of this study is to optimise the phenolic extraction process by DIE and to examine the antioxidant activities. The extraction process parameters were observed by central composite design. The antioxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The results showed that the optimal extraction condition is extract time of 90 min, temperature of 45°C, solid-liquid ratio of 1:20 g/mL, vacuum degree of - 0.08 MPa, ethanol concentration of 60%, while the phenolic content was 5.4%. The phenolic extracts from Rhus verniciflua Stokes bark had better antioxidant activities; the antioxidant activity (IC50) of the DIE was 20 µg/mL by the DPPH method. The reducing power of the phenolic extracts was significantly related to their total phenolic content (R = 0.9903). The results presented show that the DIE method is an effective method for polyphenols extraction.

Prediction of fatty acid composition in Camellia oleifera oil by near infrared transmittance spectroscopy (NITS).

Under the serious circumstances of Camellia oleifera adulteration, the accurate examination for quality trait of C. oleifera oil is extremely urgent. For rapid determination of FA composition in C. oleifera oil, the feasibility of NITS was first studied. The quantitative models for FA were built based on PLS regression. NITS spectra is able to accurately predict for oleic, linoleic, and palmitic acids (R(cv)>0.844, R(2)>0.886). R(cv) are 0.91987, 0.95755, and 0.84447, and R(2) are 0.9424, 0.9682, 0.8862 for NITS models of oleic, linoleic, and palmitic acids, respectively. But models for stearic and unsaturated acids are less accurate, with values of R(cv) from 0.67440 to 0.69114, and R(2) from 0.6834 to 0.7587. These results indicate that NITS will have potential to be used in predicting FA composition of C. oleifera oil.

Purification and characterization of a novel ~18 kDa antioxidant protein from Ginkgo biloba seeds.

Ginkgo biloba seeds are widely used as a food and traditional medicine in China. In the present study, a novel antioxidant protein named GBSP was purified from Ginkgo biloba seeds. The protein (GBSP) was purified by homogenization of Ginkgo biloba seed powder in saline solution, 70% ammonium sulphate precipitation, filtration on a DEAE-Cellulose52 anion exchange column, gel filtration on a Sephadex G-50 column, and preparative chromatography on a C(18) column using RP-HPLC. GBSP showed an apparent molecular weight of 18 kDa by SDS-PAGE and MALDI-TOF/MS analyses. The amino acid sequence obtained by MALDI-TOF/TOF MS analysis showed GBSP was a novel protein, as no matching protein in was found the database. The protein exhibited significant antioxidant activities against free radicals such as DPPH, ABTS and superoxide anion and showed higher activity than α-tocopherol in a linoleic acid emulsion assay system. Furthermore, GBSP exhibited notable reducing power and a strong chelating effect on Cu(2+) and Fe(2+). Therefore, the present study demonstrates, for the first time, that this novel protein from Ginkgo biloba seeds is an excellent antioxidant.

Hepatoprotective effects of polyprenols from Ginkgo biloba L. leaves on CCl4-induced hepatotoxicity in rats.

The hepatoprotective effects of polyprenols from Ginkgo biloba L. leaves were evaluated against carbon tetrachloride induced hepatic damage in Sprague-Dawley rats. The elevated levels of serum ALT, AST, ALP, ALB, TP, HA, LN, TG, and CHO were restored towards normalization significantly by GBP in a dose dependent manner. The biochemical observations were supplemented with histopathological examination of rat liver sections. Meanwhile, GBP also produced a significant and dose-dependent reversal of CCl(4)-diminished activity of the antioxidant enzymes and reduced CCl(4)-elevated level of MDA. In general, the effects of GBP were not significantly different from those of the standard drug Essentiale.