Preventive Effect and Mechanism of Anthocyanins from Aronia Melanocarpa Elliot on Hepatic Fibrosis Through TGF-ß/Smad Signaling Pathway.

In order to explore the effect and mechanism of Aornia mealnocarpa Elliot anthocyanins (AMA) at the cellular level on hepatic fibrosis (HF), molecular docking, RT-PCR and Western Blotting were used to explore the molecular mechanism and the effects of different doses AMA on HSC-T6 cells by TGF-ß1 induction. The results showed that the binding energy of anthocyanins on TGF-ß1 (PDB ID: 3KFD) was in the range of -9.5 to 8.6 kcal/mol, with good low energy parameters and binding positions. AMA could effectively inhibit the expressions of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total serum bilirubin (TSB), and improved the expressions of total protein (TP) and albumin (ALB). RT-PCR and Western bloting results showed that AMA could inhibited the secretion of inflammatory cytokines IL-1, IL-6, TNF-α and COX-2, and inhibit the expression of TGF-ß1, P-Smad2, α-SMA and Collagen I in TGF-ß /Smad signaling pathway. This study revealed the AMA's inhibition effects and mechanism of malignant biological behavior of HSC-T6 cells, in order to provide theoretical basis for the prevention and treatment of HF by Aronia melanocarpa Elliot.

Ganoderma immunomodulatory proteins: mushrooming functional FIPs.

Fungal immunomodulatory protein (FIP) is a novel functional protein family with specific immunomodulatory activity identified from several macro-fungi. A variety of biological activities of FIPs have been reported, such as anti-allergy, anti-tumor, mitogenic activity, and immunomodulation. Among all known FIPs, the firstly discovered FIP was isolated from Ganoderma lucidum, and most FIP members were from Ganoderma genus. Compared with other FIPs, Ganoderma FIPs possess some advantageous bioactivities, like stronger anti-tumor activity. Therein, gene sequences, protein structural features, biofunctions, and recombinant expression of Ganoderma FIPs were summarized and addressed, focusing on elucidating their anti-tumor activity and molecular mechanisms. Combined with current advances, development potential and application of Ganoderma FIPs were also prospected. KEY POINTS: • More than a dozen of reported FIPs are identified from Ganoderma species. • Ganoderma immunomodulatory proteins have superior anti-tumor activity with promising prospects and application. • Current review comprehensively addresses characterization, biofunctions, and anti-tumor mechanisms of Ganoderma FIPs.

Effect of pectin oligosaccharides supplementation on infant formulas: The storage stability, formation and intestinal absorption of advanced glycation end products.

Pectin oligosaccharides with a molecular weight greater than 700 Da was obtained from the pomace of kiwi (Actinidia arguta). Based on characteristics analysis and inhibitory activity of advanced glycation end products (AGEs) formation in vitro, the target pectin oligosaccharides was added to infant formulas and then subjected to accelerated storage. Results showed that pectin oligosaccharides supplementation inhibited the browning of infant formulas and glassy transition of lactose, and slowed down the increase of water activity under accelerated storage conditions. Pectin oligosaccharides also inhibited the formation of AGEs in infant formulas, such as 5-(hydroxymethyl)furfural, Nε-carboxymethyl-lysine, Nε-carboxyethyl-lysine, methylglyoxal hydromidazolones, glyoxal hydromidazolones, glyoxal-lysine dimer, methylglyoxal-lysine dimer and pyrraline. Besides, permeability studies using Caco-2 cell monolayer also showed that pectin oligosaccharides supplementation inhibited the intestinal absorption of AGEs, especially 5-(hydroxymethyl)furfural, Nε-carboxymethyl-lysine, Nε-carboxyethyl-lysine and glyoxal hydromidazolones. These results provide a reliable theoretical basis for the application of pectin oligosaccharides in infant formulas.

Anthocyanins from Aronia melanocarpa Induce Apoptosis in Caco-2 Cells through Wnt/ß-Catenin Signaling Pathway.

Colorectal cancer (CRC) is one of the most common malignant tumors in the world. In this study, the Caco-2 in vitro cell model was used to study the effect and mechanism of Aronia melanocarpa (Michx.) Elliott anthocyanins (AMA) on colon cancer. The experimental results showed that the binding energy of anthocyanins on ß-catenin was in the range of -5.92 to 4.95 kcal/mol, with good low energy parameters and binding positions. AMA can inhibit cell proliferation and cause cell cycle arrest. RT-PCR and Western blot results showed that AMA can reduce cytoplasmic ß-catenin and inhibit the expression of related proteins in Wnt/ß-catenin signaling pathway. This study revealed the AMA inhibitory effect and mechanism of malignant biological behavior of Caco-2 cells, in order to provide theoretical basis for the prevention and treatment of colon cancer by Aronia melanocarpa (Michx.) Elliott.

Pectin oligosaccharides from fruit of Actinidia arguta: Structure-activity relationship of prebiotic and antiglycation potentials.

Pectin oligosaccharides (POSs) have prebiotic and antiglycation activities in vitro, but the specific structure-activity relationship is unclear. In this study, POSs were obtained by enzymatic and ultrasound-assisted enzymatic degradation of pectin polysaccharide (PPS), respectively. Based on the chemical characterization, the antiglycation in vitro and prebiotic activities of POSs were compared and the structure-activity relationship was studied. The results showed that the antiglycation activity of POSs in vitro was proportional to the galacturonic acid content and GalA:Rha molar ratios except for the low molecular weight POSs (LM-POSs), and inversely proportional to its branching degree, such as Ara:Rha and Gal:Rha molar ratios. In addition, it was also found that the prebiotic activity of POSs was positively correlated with Ara:Rha and Gal:Rha molar ratios in molecule composition and the neutral sugar content, especially galactose and arabinose. The degree of esterification (DE) was less important for both antiglycation and prebiotic activity of POSs. These results provided an important theoretical basis for POSs application in food.

Characterization of polysaccharide fractions from fruit of Actinidia arguta and assessment of their antioxidant and antiglycated activities.

A novel cell-wall polysaccharides (AAPs) were extracted from the fruits of Actinidia arguta and separated into four parts which were named water-eluted polysaccharide (WPS), salt-eluted polysaccharide (SPS)-1, SPS-2 and SPS-3. The monosaccharide composition and structural analysis showed that SPS-3 and SPS-2 were homogalacturonan (HG)-rich pectin, SPS-1 was rhamnogalacturonan (RG)-rich pectin and WPS was starch-like polysaccharides. All four kinds of polysaccharides displayed the ability to scavenge free radicals, chelate iron ion, inhibit lipid peroxidation and inhibit protein glycation, but SPS was obviously stronger than WPS. Especially SPS-3 displayed the strongest antioxidant and anti-glycated activities. In addition, the inhibitory effect of AAPs on AGEs formation is attributed to the inhibitory activity on the production of protein carbonyl group and the protective effects on the protein thiol group but not the scavenging capacity of dicarbonyl compounds, suggesting that its mechanisms of antiglycated effects may be of concern to their antioxidant activities.

Anthocyanins Delay Ageing-Related Degenerative Changes in the Liver.

Liver ageing is a significant risk factor for chronic liver diseases. Anthocyanin is a food additive that has previously shown efficacy in increasing longevity. Here, we tested whether anthocyanins could protect young mice from accelerated ageing of the liver. Kunming mice were injected with D-galactose to accelerate ageing and were given 20 or 40 mg/kg anthocyanins as an intervention. After eight weeks, whole liver function and structure were evaluated, and the expression levels of genes involved in the DNA damage signalling pathway were assessed by Western blot analysis. Anthocyanins delayed the reduction of the liver index (p < 0.05), hepatic tissue injury and fibrosis. Anthocyanins also maintained the stability of the redox system (GSH-PX, T-SOD and MDA) in plasma and liver structures (p < 0.001) and reduced the levels of inflammatory factors (IL-1, IL-6 and TNF-α) in the liver (p < 0.05). Moreover, the expression levels of sensors (ATM and ATR), mediators (H2AX and γ-H2AX) and effectors (Chk1, Chk2, p53 and p-p53) in the DNA damage signalling pathway were all reduced. Anthocyanins could be widely used in the field of health products to slow ageing-related deterioration of liver function and structure by inhibiting DNA damage.

Anthocyanins from Black Chokeberry (Aroniamelanocarpa Elliot) Delayed Aging-Related Degenerative Changes of Brain.

Aging is the greatest risk factor for most neurodegenerative diseases, which is associated with decreasing cognitive function and significantly affecting life quality in the elderly. Computational analysis suggested that 4 anthocyanins from chokeberry fruit increased Klotho (aging-suppressor) structural stability, so we hypothesized that chokeberry anthocyanins could antiaging. To explore the effects of anthocyanins treatment on brain aging, mice treated with 15 or 30 mg/kg anthocyanins by gavage and injected D-galactose accelerated aging per day. After 8 weeks, cognitive and noncognitive components of behavior were determined. Our studies showed that anthocyanins blocked age-associated cognitive decline and response capacity in senescence accelerated mice. Furthermore, mice treated with anthocyanins-supplemented showed better balance of redox systems (SOD, GSH-PX, and MDA) in all age tests. Three major monoamines were norepinephrine, dopamine, and 5-hydroxytryptamine, and their levels were significantly increased; the levels of inflammatory cytokines (COX2, TGF-ß1, and IL-1) transcription and DNA damage were decreased significantly in brains of anthocyanins treated mice compared to aged models. The DNA damage signaling pathway was also regulated with anthocyanins. Our results suggested that anthocyanins was a potential approach for maintaining thinking and memory in aging mice, possibly by regulating the balance of redox system and reducing inflammation accumulation, and the most important factor was inhibiting DNA damage.

Pectin Penta-Oligogalacturonide Suppresses Intestinal Bile Acids Absorption and Downregulates the FXR-FGF15 Axis in High-Cholesterol Fed Mice.

Haw pectin penta-oligogalacturonide (HPPS), purified from the hydrolysates of haw pectin, has important role in decreasing hepatic cholesterol accumulation and promoting bile acids (BA) excretion in the feces of mice fed a high-cholesterol diet (HCD). However, the mechanism is not clear. This study aims to investigate the effects of HPPS on BA reabsorption in ileum and biosynthesis in liver of mice. Results showed that HPPS increased fecal BA output by approximately 110%, but decreased ileal BA and the total BA pool size by approximately 47 and 36%, respectively, compared to HCD. Studies of molecular mechanism revealed that HPPS significantly decreased the mRNA and protein levels of farnesoid X receptor (FXR) in the small intestine of mice and inactivated the fibroblast growth factor 15 (FXR-FGF15) axis, which increased the mRNA and protein levels of CYP7A1 by approximately 204 and 104%, respectively, compared to HCD. Interestingly, the mRNA and protein levels of apical sodium-dependent bile acid transporter (ASBT) in the small intestine were approximately 128 and 73% higher in HPPS-fed mice than those in HCD-fed mice, respectively. However, no significant difference was detected for ASBT expression between HCD group and BA sequestrant cholestyramine group. These findings indicate that HPPS can suppress intestinal BA reabsorption and promoting hepatic BA biosynthesis. We speculated that HPPS could be ASBT competitive inhibitor rather than BA sequestrant in inhibiting BA reabsorption in ileum and improving cholesterol metabolism.

Pectin penta-oligogalacturonide reduces cholesterol accumulation by promoting bile acid biosynthesis and excretion in high-cholesterol-fed mice.

Haw pectin penta-oligogalacturonide (HPPS) has important role in improving cholesterol metabolism and promoting the conversion of cholesterol to bile acids (BA) in mice fed high-cholesterol diet (HCD). However, the mechanism is not clear. This study aims to investigate the effects of HPPS on cholesterol accumulation and the regulation of hepatic BA synthesis and transport in HCD-fed mice. Results showed that HPPS significantly decreased plasma and hepatic TC levels but increased plasma high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A-I (apoA-I) levels, compared to HCD. BA analysis showed that HPPS markedly decreased hepatic and small intestine BA levels but increased the gallbladder BA levels, and finally decreased the total BA pool size, compared to HCD. Studies of molecular mechanism revealed that HPPS promoted hepatic ATP-binding cassette transporter A1 (ABCA1), ATP-binding cassette transporter G1 (ABCG1), and scavenger receptor BI (SR-BI) expression but did not affect ATB binding cassette transporter G5/G8 (ABCG5/8) expression. HPPS inactivated hepatic farnesoid X receptor (FXR) and target genes expression, which resulted in significant increase of cholesterol 7α-hydroxylase 1 (CYP7A1) and sterol 12α-hydroxylase (CYP8B1) expression, with up-regulations of 204.2% and 33.5% for mRNA levels, respectively, compared with HCD. In addition, HPPS markedly enhanced bile salt export pump (BSEP) expression but didn't affect the sodium/taurocholate co-transporting polypeptide (NTCP) expression. In conclusion, the study revealed that HPPS reduced cholesterol accumulation by promoting BA synthesis in the liver and excretion in the feces, and might promote macrophage-to-liver reverse cholesterol transport (RCT) but did not liver-to-fecal RCT.

Polyclonal antibodies from hen egg yolk (IgY) with hydrolysis activity.

Polyclonal catalytic antibodies (abzymes) play an important role in immunology research. In this study, we report polyclonal antibodies IgYs isolated from chicken egg yolk with hydrolysis activity for the first time. The IgYs were raised in hens using HNPBV [4-(hydroxy (naphthalen-2-yloxy) phosphoryl) butanoic acid] attached to BSA (Bovine serum albumin) as an immunogen. Anti-(HNPBV-BSA) IgYs were isolated from yolks of the eggs laid using a two-step salt precipitation and one-step gel filtration protocol. NA (naphthalen-2-yl acetate) was selected as the substrate and the hydrolysis reaction of the IgYs for it was examined. The result reveals that the rate of the hydrolysis reaction is higher (Kcat/K (uncat) approximately 2x10(4)). The purified IgYs were digested with pepsin and the smaller fragment (Fab') with specific antigen binding properties was produced. The research indicates that the enzymatic properties of Fab' are similar to IgYs. The catalytic activity of the IgYs was further determined by measuring the rate of hydrolysis of NA in the presence of inhibitor. These findings show that chicken egg is an excellent donor for polyclonal catalytic antibodies.