A novel near-infrared fluorescence off–on probe, (E)-3,3-dimethyl-1-propyl-2-(2-(6-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyloxy)-2,3-dihydro-1H-xanthen-4-yl)vinyl)-3H-indolium (1), is developed and applied to benzoyl peroxide (BPO) detection in real samples and fluorescence imaging in living cells and zebrafish. When arylboronate as the recognition unit is connected to a stable hemicyanine skeleton, the probe is readily prepared, which exhibits superior analytical performance, such as near-infrared fluorescence emission over 700 nm and high sensitivity with a low detection limit of 47 nM. Upon reaction with BPO, phenylboronic acid pinacol ester is oxidized, followed by hydrolysis and 1,4-elimination of o-quinone methide to release fluorophore. In addtion, the probe displays high selectivity toward BPO over other common substances, which makes it of great potential use in quantitative and simple detection of BPO in wheat flour and antimicrobial agent. More importantly, the probe has been successfully demonstrated for monitoring BPO in living HeLa cells and zebrafish. The probe with superior properties could be of great potential use in other biosystems and in vivo studies.Keywords: benzoyl peroxide; fluorescent probe; imaging analysis; zebrafish;

A new near-infrared fluorescence off–on probe was developed and applied to fluorescence imaging of carboxylesterase in living HepG-2 cells and zebrafish pretreated with pesticides (carbamate, organophosphorus, and pyrethroid). The probe was readily prepared by connecting (4-acetoxybenzyl)oxy as a quenching and recognizing moiety to a stable hemicyanine skeleton that can be formed via the decomposition of IR-780. The fluorescence off–on response of the probe to carboxylesterase is based on the enzyme-catalyzed spontaneous hydrolysis of the carboxylic ester bond, followed by a further fragmentation of the phenylmethyl unit and thereby the fluorophore release. Compared with the only existing near-infrared carboxylesterase probe, the proposed probe exhibits superior analytical performance, such as near-infrared fluorescence emission over 700 nm as well as high selectivity and sensitivity, with a detection limit of 4.5 × 10–3 U/mL. More importantly, the probe is cell membrane permeable, and its applicability has been successfully demonstrated for monitoring carboxylesterase activity in living HepG-2 cells and zebrafish pretreated with pesticides, revealing that pesticides can effectively inhibit the activity of carboxylesterase. The superior properties of the probe make it of great potential use in indicating pesticide exposure.Keywords: carboxylesterase; fluorescent probe; imaging analysis; pesticide;

This study was to develop a novel strategy for the simultaneous consumption of soluble soybean polysaccharides (SSPS) to enhance the absorption of genistein and its protective effects against high L-carnitine-induced hepatic injury in mice. UPLC-qTOP/MS measurements showed that SSPS observably increased the urinary concentration of genistein and its metabolites in mice. The mice fed with 3% L-carnitine water for 12 weeks experienced a disturbance of the hepatic lipid metabolism, oxidative stress and inflammation, which was evidenced by abnormal TC, LDL, RAHFR and MDA levels, unusual AST, ALT, ALP, SOD and GSP-Px activities, and increased IF-1, IF-6 and TNF-α expressions. Interestingly, the co-supplementation of SSPS and genistein was capable of regulating these imbalances more effectively than the administration of SSPS or genistein alone, which was also confirmed by histological observations of the mouse liver. These findings suggest that the co-ingestion of SSPS and genistein is a feasible strategy for improving liver protection in mice.

This study was to develop a novel strategy for the simultaneous consumption of soluble soybean polysaccharides (SSPS) to enhance the absorption of genistein and its protective effects against high L-carnitine-induced hepatic injury in mice. UPLC-qTOP/MS measurements showed that SSPS observably increased the urinary concentration of genistein and its metabolites in mice. The mice fed with 3% L-carnitine water for 12 weeks experienced a disturbance of the hepatic lipid metabolism, oxidative stress and inflammation, which was evidenced by abnormal TC, LDL, RAHFR and MDA levels, unusual AST, ALT, ALP, SOD and GSP-Px activities, and increased IF-1, IF-6 and TNF-α expressions. Interestingly, the co-supplementation of SSPS and genistein was capable of regulating these imbalances more effectively than the administration of SSPS or genistein alone, which was also confirmed by histological observations of the mouse liver. These findings suggest that the co-ingestion of SSPS and genistein is a feasible strategy for improving liver protection in mice.

This study was designed to investigate the preventive effects of Red Fuji apple peel polyphenolic extract (APP) on vascular endothelial dysfunction and liver injury in mice fed a high choline diet. The mice were fed 3% dietary choline in drinking water for 8 weeks and displayed vascular dysfunction and liver damage (p < 0.01). The administration of APP at 600 and 900 mg per kg bw significantly elevated serum NO, HDL and 6-Keto-PGF1a levels and lowered serum TC, TG, LDL, ET-1 and TXB2 levels in the HC-fed mice. Besides, APP also caused the reduction of AST, ALT activities and MDA, CRP, TNF-α levels, and increased the hepatic GSH-Px and SOD activities of the HC-fed mice. Furthermore, the histopathology of the liver by conventional H&E and oil red O staining confirmed the liver steatosis induced by a choline diet and the hepatoprotective effect of APP. The experiment results indicated that the polyphenolic extract from apple peel might be regarded as a preventive and therapeutic product for the amelioration of HC diet-induced vascular dysfunction and hepatic injury.

•RSM optimization of TP from EAE of Ulmus pumila L. barks.•EAE, UAE and CHE were compared for extraction of TP.•The antioxidant potential and phenolic profile of the extracts were characterized.•EAE afforded higher yield of phenolics and antioxidant activity than UAE and CHE.This study aimed to optimize trienzyme-assisted extraction (EAE) conditions for total polyphenols (TP) from Ulmus pumila barks (UPB). Response surface methodology (RSM) was used to optimize EAE conditions including pH, temperature, and time. The extraction efficiency of three extraction procedures on the TP yield, antioxidant activities and chemical composition of UPB extracts was also compared and characterized. Our results showed that the maximum extraction yield of TP was 16.04 ± 0.38 mg gallic acid equivalents/g dry weight (GAE/g DW) under the optimum EAE conditions (pH = 4.63, 52.6 °C and 62 min). Meanwhile, the EAE gave a higher extraction yield of TP and then a greater in vitro antioxidant capacity compared with those obtained from both ultrasound-assisted extraction (UAE) and conventional heat extraction (CHE). In addition, seven polyphenolic compounds were validated by high-performance liquid chromatography analysis in the extracts at the optimized conditions. The results of this study further confirmed that EAE could be explored as a state-of-the-art environmentally friendly technology for recovering optimum amounts of antioxidant polyphenols from plant sources.

•A novel nanoparticle system was developed from Lysozyme and ASKP-3 through self-assembly.•Encapsulation of curcumin in the Ly/ASKP-3 NPs enhanced its chemical stability.•Curcumin-loaded NPs exhibited stronger antitumor activity toward HepG2 cells than free curcumin.•Curcumin-loaded NPs significantly enhanced serum level of curcumin in mice.A facile approach to fabricate nanoparticles (NPs) via self-assembly of lysozyme (Ly) and purified Artemisia sphaerocephala Krasch seed polysaccharide (ASKP-3) was developed for the first time to encapsulate and protect curcumin. The nanoparticle size and morphology were characterized. The results showed that Ly/ASKP-3 NPs were spherical with narrow particle size distribution. Curcumin-loaded NPs greatly increased curcumin stability in phosphate buffer at physiological pH. The cancer cell viability assay indicated that encapsulated curcumin exhibited higher antitumor potential than free curcumin. Curcumin-loaded NPs was effectively endocytosed by HepG2 cells, resulting in enhanced apoptosis in cancer cells as compared to free curcumin. Furthermore, the curcumin-loaded NPs significantly increased the serum level of curcumin in mice after oral administration. Overall, our results demonstrated that nanosized proteins/polysaccharides complex could be used to protect labile lipophilic compounds such as curcumin to enhance their bioavailability in vivo, supporting potential application of curcumin-loaded proteins/polysaccharides complex in functional foods.Download high-res image (173KB)Download full-size image

The present study was designed to investigate the protective effects of Ilex Kuding tea polysaccharides (IKTP) on high fructose (HF)-induced liver injury and vascular endothelial dysfunction in mice. IKTP were identified as acidic heteropolysaccharides by FT-IR and HPLC. Healthy male Kunming mice fed 20% fructose in drinking water for 8 consecutive weeks significantly displayed dyslipidemia, hepatic steatosis, oxidative stress and vascular endothelial dysfunction. However, continuous administration of IKTP at 200, 400 and 800 mg per kg bw in HF-fed mice could prevent the damage caused by HF-diets, especially at dosages of 400 and 800 mg per kg bw (p < 0.01). IKTP significantly reduced the HF-induced elevation of the serum TC, TG, LDL-C, TXA2 and ET-1 levels, as well as AST and ALT activities, while markedly increased the HF-induced decline of HDL-C, PGI2 and eNOS levels in the serum compared to HF-fed mice. Meanwhile, the hepatic MDA level was lowered while SOD and GSH-Px activities were increased in IKTP-treated mice, compared to HF-fed mice. Histopathology of the liver and cardiovascular aortic by H&E or oil red O staining confirmed the liver steatosis and the vascular injury induced by HF-diets and the protective effects of IKTP. These findings suggest that HF causes oxidative damage, and IKTP alleviates liver injury and vascular endothelial dysfunction.

The aim of this study was to investigate the molecular mechanism underlying the immunomodulatory effect of the purified Artemisia sphaerocephala Krasch seed polysaccharide (ASKP-1) in RAW264.7 macrophages. Chemical characteristic analysis revealed that ASKP-1 consisted of 14.1% mannose, 56.9% glucose and 19.6% galactose with the average molecular weight of 9.08 × 105 Da and the mixed glycan backbone structure containing 1→4)-Glcp (39.8%), 1→6)-Galp (18.8%), 1→3,6)-Manp (19.6%), 1→)-Glcp (10.8%), 2→6)-Manp (4.0%) and 2→3,5)-Araf (7.0%). In vitro studies showed that ASKP-1 markedly induced the release of cytotoxic molecules (NO and ROS) and secretion of the cytokines (TNF-α, INF-β, and IL-6) and significantly enhanced the phagocytosis of RAW264.7 macrophages. Furthermore, TLR4 was found to be a recognized target of ASKP-1 and its related mitogen-activated protein (MAPK) and phosphoinositide 3-kinase (PI3K)/Akt, including phosphorylated ERK, JNK, p38 and Akt, were rapidly activated by ASKP-1 in RAW264.7 macrophages. Moreover, ASKP-1 was found to cause the nuclear translocation of the nuclear factor NF-κB subunit p65 and the degradation of IκB-α in RAW264.7 macrophages. All these findings suggest that MAPK, PI3K/Akt and NF-κB pathways are involved in ASKP-1-induced macrophage activation, and ASKP-1 is a potential immunomodulating function food.

Deshipu stachyose granules (DSG) is a mixture of α-galacto-oligosaccharides derived from the dietary roots of Lycopus lucidus Turcz. Our previous study showed that DSG could improve the faecal microbial composition, and facilitate intestinal peristalsis and fecal excretion in mice. This study was designed to investigate the effect of DSG on gut microbiota and bowel function in humans. Two human intervention studies were conducted. In the first study, 100 healthy adults were treated without or with 5 g per day of DSG for 14 days. The microbiota composition in fecal samples was quantitatively analyzed before and after DSG supplementation. We found that DSG consumption significantly elevated the fecal bifidobacteria and lactobacilli levels, and also decreased the fecal Clostridium perfringens concentration. In the second study, 103 constipated patients were treated with 5 g per day of placebo or DSG for 30 days, and subsequently subjected to bowel function evaluation. As a result, dietary intake of DSG effectively improved the bowel function of constipated patients, as evidenced by the increased defecation frequency, softer stools and easier defecation. Moreover, clinical safety assessment showed that DSG at 5 g per day did not cause significant adverse effects in both healthy and constipated volunteers. In conclusion, DSG at 5 g d−1 beneficially modulated the gut microbiota in healthy adults and potently improved the bowel function of constipated patients without consequent adverse events. This study suggests that DSG holds promising potential for safe treatment of functional constipation.

Vascular insulin resistance and oxidative stress contribute to endothelial dysfunction and hypertension. The present study investigated whether chronic treatment with purified Tartary buckwheat flavonoids fraction (TBF) prevents the development of hypertension via improving vascular insulin sensitivity and reducing oxidative stress. Six-week-old male spontaneously hypertensive rats (SHRs) and their normotensive Wistar-Kyoto (WKY) control rats were subjected to different dosages of TBF for 8 weeks. Blood pressure, mesenteric arteriolar vasorelaxation, superoxide anion (O2−) generation, NAD(P)H oxidase activity, and insulin-stimulated Akt/endothelial nitric oxide synthase (eNOS) activation and nitric oxide (NO) production were determined. The SHRs had higher systolic blood pressure, systemic insulin resistance, and impaired vasodilator actions of insulin and the insulin signaling pathway in mesenteric arterioles when compared with the WKY rats. TBF treatment at a dosage of 100 mg kg−1 day−1 significantly reduced systolic blood pressure and increased vasodilator response to insulin in the SHRs. Additionally, TBF treatment significantly reduced phosphorylation of insulin receptor substrate 1 (IRS-1) at serine 307 and increased insulin-stimulated Akt/eNOS activation in the SHRs. Furthermore, TBF treatment reduced the overproduction of basal O2− in association with a reduction of NAD(P)H oxidase activity in mesenteric arterioles of the SHRs. Finally, quercetin was identified as the predominant active component of TBF in attenuating the development of hypertension with regard to reducing vascular oxidative stress, regulating the vascular insulin signaling pathway and restoring vasodilator response to insulin in the SHRs. In conclusion, TBF possesses protective effects against hypertension through attenuating vascular insulin resistance and oxidative stress.

•PEP is the crude heteropolysaccharides isolated from Pleurotus eryngii.•PEP-1 and PEP-2 were purified from PEP by Cellulose DEAE-52 and Sephadex G-100.•High molecular mass PEP-2 exhibits more potent anticancer effect than PEP-1.•ROS was a critical mediator in PEP-2 induced HepG-2 cellular apoptosis.•PEP-2 is a good candidate for dietary supplement and therapeutic application in cancer.This study was designed to investigate the chemical characterization and antitumor effects of Pleurotus eryngii polysaccharides (PEP). The crude PEP was fractionated into two fractions, namely PEP-1 and PEP-2. HPLC analysis showed that PEP-1 and PEP-2 were heteropolysaccharides mainly composed of glucose with the average molecular weights of 2.54 × 104 Da (PEP-1) and 4.63 × 105 Da (PEP-2), respectively. High molecular mass PEP-2 was shown to exhibit stronger growth inhibition against human hepatoblastoma HepG-2 cells in comparison with PEP-1. Flow cytometric analysis showed that PEP-2 exerted a stimulatory effect on apoptosis of HepG-2 cells, and induced the cell-cycle arrest at the S-phase, with the observation of intracellular ROS production. These findings suggest that the polysaccharides, especially PEP-2, are very important nutritional ingredients responsible for the anticancer health benefits of P. eryngii.

Isoorientin (ISO), a natural flavonoid, has been found to have multiple biological properties. In the present study, obese mice with high-fructose (HF)-induced liver injury were used to investigate the hepatoprotective effects of ISO. The results showed that ISO significantly reduced the serum lipid parameters in mice fed 20% HF water. Meanwhile, ISO appeared to alleviate HF-induced lipid metabolic disorders by increasing the serum levels of apo-A1 and decreasing the serum apoB levels, apoB/apo-A1 ratio, and FAS activity in the liver. ISO also remarkably ameliorated HF-induced hepatic oxidative injury and inflammation by decreasing ALT, AST, and ALP levels; enhancing antioxidant enzyme activities; and inhibiting inflammatory cytokine (TNF-α, IL-1, IL-6) release. Histopathology of liver stained by H&E and Oil Red O showed the liver steatosis and oxidative injury after HF treatment and the protective effect of ISO. Furthermore, aortic pathology observation found that ISO had a protective effect on the vascular endothelium. This is the first report that ISO efficiently inhibited HF-induced hyperlipidemia and liver injury by ameliorating lipid metabolism, enhancing the antioxidant defensedefense system, and regulating the secretion of inflammatory cytokines.

Dietary supplementation of soy stachyose or genistein is known to be of hepatoprotective health interest. This study showed that co-administration of genistein and stachyose caused stronger inhibition on abnormal weight gain and liver fat accumulation by decreasing fatty acid synthetase expression and balancing disorderly lipid metabolism than that of genistein or stachyose alone in high-fructose (HF) diet-fed mice. Furthermore, the production of malonaldehyde and carbonyl derivatives of proteins was also more effectively inhibited by co-treatment of genistein and stachyose, and thereby glutathione peroxidase and superoxide dismutase activities were elevated in HF-fed mice. Moreover, genistein in combination with stachyose was more effective to reduce the impact of HF on the serum markers of liver damage by inhibiting inflammatory cytokine release than stachyose or genistein alone in mice. The potential mechanism was that stachyose enhanced absorption of genistein in HF-fed mice by oral supplementation of genistein together with stachyose. These findings indicate that co-ingestion of stachyose and genistein may serve as a novel strategy for hepatic protection.

The present study was designed to evaluate the effects of selenium-containing tea polysaccharides (Se-GTP) from a new variety of selenium-enriched Ziyang green tea against high fructose (HF)-induced insulin resistance and hepatic oxidative stress in mice. Healthy male Kunming mice were fed 20% high fructose water and administered 200, 400 and 800 mg per kg bw Se-GTP for 8 weeks. Mice fed HF in drinking water displayed significant insulin resistance, hepatic steatosis and oxidative stress observed by hyperglycemia and hyperinsulinemia, as well as increases in hepatic non-esterified fatty acid (NEFA) and malonaldehyde (MDA). The administration of Se-GTP at 400 and 800 mg per kg bw significantly improved insulin sensitivity, and reduced liver steatosis and oxidative stress damage, and brought back the antioxidants and hepatic lipids towards near-normal values. In the oral glucose tolerance test, the administration of Se-GTP at 400 and 800 mg per kg bw had reduced plasma glucose concentrations after 30 min of glucose loading in HF-fed mice, suggesting that Se-GTP improved glucose intolerance. Histopathological examination indicated that the impaired pancreatic/hepatic tissues were effectively restored in HF-fed mice following the Se-GTP treatment. This is the first report showing that Se-GTP can ameliorate the high fructose-induced insulin resistance and hepatic oxidative injury.

This study was designed to investigate the protective effects of D-Chiro-Inositol (DCI) enriched tartary buckwheat extract (DTBE) against high fructose (HF) diet-induced hyperglycemia and liver injury in mice. HPLC analysis revealed that the content of DCI present in purified DTBE was 34.06%. Mice fed 20% fructose in drinking water for 8 weeks significantly displayed hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis and oxidative stress (p < 0.01). Continuous administration of DTBE in HF-fed mice dose-dependently reduced the HF-induced elevation of body weight, serum glucose, insulin, total cholesterol (TC), total triglycerides (TG) and low density lipoprotein cholesterol (LDL-C) levels, as well as serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), C-reactive protein (CRP) and lactate dehydrogenase (LDH) activities, while the HF-induced decline of serum high density lipoprotein-cholesterol (HDL-C) levels could be markedly elevated in the mice. Meanwhile, DTBE also dose-dependently increased the hepatic total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) activities, and decreased hepatic malonaldehyde (MDA) levels, relative to HF-treated mice. Histopathology of H&E and Oil Red O staining confirmed liver injury induced by a HF diet and the hepatoprotective effect of DTBE. These findings are the first to demonstrate that the intake of DTBE may be a feasible preventive or therapeutic strategy for HF diet-induced hyperglycemia, hepatic steatosis and oxidative injury.

This study was performed to investigate the liver and vascular changes in high trimethylamine-N-oxide (TMAO) diet-fed mice, and the possible vasoprotective and hepatoprotective effects of purified tartary buckwheat flavonoid fraction (TBF). HPLC analysis revealed that the content of rutin and quercetin presented in TBF was 53.6% and 37.2%, respectively, accounting for 90.8% of TBF. Mice fed 1.5% TMAO in drinking water for 8 weeks significantly displayed vascular dysfunction and liver damage (p < 0.01). The administration of TBF at 400 and 800 mg per kg bw significantly elevated plasma NO and eNOS concentrations, and serum HDL-C and PGI2 levels, and lowered serum TC, TG, LDL-C, ET-1 and TX-A2 levels of TMAO-fed mice. TBF also reduced serum AST and ALT activities, and hepatic NEFA and MDA levels, and increased the hepatic GSH-Px and SOD activities in TMAO-fed mice, which were consistent with the observations of the histological alterations of the liver. This report firstly showed that dietary TMAO might cause liver damage and TBF prevented TMAO-induced vascular dysfunction and hepatic injury.

The present study was conducted to explore the protective effects of myricetin (MYR) purified from Hovenia dulcis Thunb. against vascular endothelial dysfunction and liver injury in mice fed with 3% dietary choline water. MYR was shown to possess strong scavenging activities against DPPH˙, HO˙, and O2˙− and ferric-reducing antioxidant power in vitro. Mice fed 3% dietary choline water for 8 weeks significantly displayed vascular endothelial dysfunction and liver oxidative stress (p < 0.01). Furthermore, continuous administration of MYR at 400 and 800 mg per kg bw in choline-fed mice could significantly decrease the high choline diet-induced elevation of serum total cholesterol (TC), total triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), endothelin 1 (ET-1) and thromboxane A2 (TXA2) levels as well as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, while the choline-induced decline of serum high density lipoprotein-cholesterol (HDL-C), endothelin nitric oxide synthase (eNOS), nitric oxide (NO) and prostaglandin I2 (PGI2) levels could be markedly elevated in mice (p < 0.05, p < 0.01). Meanwhile, MYR at 400 and 800 mg per kg bw also increased hepatic total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) activities and decreased hepatic malonaldehyde (MDA) and non-esterified fatty acid (NEFA) levels in mice, relative to choline-treated mice (p < 0.05, p < 0.01). These results together with conventional haematoxylin and eosin (H&E) and Oil Red O staining observation of the liver and vascular tissues suggested that MYR exerted a significant protective role against high choline diet-induced endothelial dysfunction and liver injury in mice. This is the first report showing that high intake of dietary choline can induce liver damage and that MYR can ameliorate choline-induced vascular endothelial dysfunction and liver injury.

The present study reports the phenolic profiles and antioxidant and hepatoprotective properties of Red Fuji apple peel polyphenolic extract (APP) and its flesh polyphenolic extract (AFP) against CCl4-induced acute hepatic damage in mice. It was found that the polyphenol and flavonoid contents of APP were significantly higher than those of AFP. APP was shown to exhibit stronger in vitro antioxidant activities than AFP in a dose-dependent manner. Administration of APP at 250 and 500 mg per kg bw to mice ahead of CCl4 injection was further shown to exhibit stronger in vivo protective effects than those of AFP and could observably antagonize the CCl4-induced increase in serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase activities and hepatic malondialdehyde levels, and prevent the CCl4-caused decrease in antioxidant superoxide dismutase and glutathione peroxidase activities, compared to CCl4-treated mice (p < 0.05). This finding demonstrates that the polyphenolic extract from apple, particularly its peel, can be explored as a chemopreventive or chemotherapeutic agent against oxidative-stress-related liver disorders.

•ZTPs are an acidic heteropolysaccharide extract from Se-enriched Ziyang green tea.•ZTPs possessed powerful scavenging ability of free radicals in vitro.•ZTPs had a prominent preventive effect against CCl4-induced hepatotoxicity in mice.•CCl4-caused histological alteration in the liver was effectively prevented by ZTPs.•ZTPs were a good candidate of dietary supplement and therapeutic application.This study was designed to investigate the hepatoprotective effects of the tea polysaccharides (ZTPs) extracted from a selenium-enriched Ziyang green tea (Camellia sinensis). ZTPs were identified as the heteropolysaccharides with glucose (31.4%), arabinose (23.5%) and galactose (21.8%) being the main constitutive monosaccharides. ZTPs displayed noteworthy scavenging effects against DPPH, OH and O2-, and high antioxidant effects in vitro, and the effects were further verified by suppressing CCl4-induced oxidative liver damage in mice at 100, 200 and 400 mg/kg BW. Administration of ZTPs in mice prior to CCl4 significantly prevented the CCl4-induced increases in serum alanine aminotransferase, aspartate aminotransferase and lactic dehydrogenase, as well as hepatic malondialdehyde level. Mice treated with ZTPs showed normal glutathione peroxidase and superoxide dismutase activities, relative to CCl4-treated group. ZTPs also prevented the CCl4-caused liver histological alteration, as indicated by histopathological evaluation. These findings demonstrate that ZTPs have protective effects against acute CCl4-induced oxidative liver damage.

Water-soluble apple peel polysaccharides (APP) and apple flesh polysaccharides (AFP) were isolated from Pink Lady fruits, and their in vitro antioxidant capacities were characterised by DPPH, HO, and O2− systems, and ferric-reducing antioxidant power assay. Oral administration of APP at 250 and 500 mg/kg bw in mice was shown to be as effective as AFP in lowering the CCl4-caused increases of serum alanine aminotransferase, aspartate aminotransferase and lactic dehydrogenase activities, and hepatic malondialdehyde level, and antagonising the decreases in antioxidant superoxide dismutase and glutathione peroxidase activities caused by CCl4 (p < 0.05). Histopathological examinations further confirmed that both APP and AFP could protect the liver from CCl4-induced histological alteration. HPLC analysis also showed similar profiles of monosaccharide composition for APP and AFP with arabinose, galactose and galacturonic acid being main component monosaccharides. All of these findings demonstrate that the extracts of both APP and AFP possess antioxidant and hepatoprotective potential.Highlights► Both APP and AFP were acidic polysaccharides from Pink Lady apple peel and flesh. ► Both APP and AFP exerted significant hepatoprotection from CCl4-induced damage in mice. ► The hepatoprotection of APP and AFP was mediated by antioxidant mechanism in mice. ► Histopathological tests show the protection from CCl4 caused histological alteration.

Highlights•HMTP is an acidic tea polysaccharide with nine monosaccharides by HPLC.•HMTP has strong free radicals-scavenging and antioxidant capacities in vitro.•Pathological test confirmed HMTP could protect CCl4-caused histological lesion.•HMTP is a potential valuable hepatoprotective functional food.This study was to examine the hepatoprotective effects of polysaccharides from green tea of Huangshan Maofeng (HMTP) against CCl4-induced oxidative damage in mice. HMTP is an acidic heteropolysaccharide with galactose (35.0%, mol.%), arabinose (28.9%) and galacturonic acid (11.3%) being the main monosaccharide components. HMTP (400 and 800 mg/kg·bw) administered orally daily for 14 days before CCl4 administration significantly reduced the impact of CCl4 toxicity on the serum markers of liver damage, alanine aminotransferase, aspartate aminotransferase, total-cholesterol and triglycerides. This method of HMTP administration also markedly restrained hepatic lipid peroxidation formation of malondialdehyde and 15-F2t isoprostanes, and elevated the antioxidant levels of hepatic glutathione and superoxide dismutase. These results together with liver histopathology indicated that HMTP exhibited hepatoprotection against CCl4-induced injury, which was found to be comparable to that of biphenyldicarboxylate. The hepatoprotective effects of HMTP may be due to both the inhibition of lipid peroxidation and the increase of antioxidant activity.

This study was aimed to isolate and characterize the raffinose family oligosaccharides (RGOs) from a novel plant source of Rehmannia glutinosa Libosch, and further evaluate whether RGOs can attenuate CCl4-induced oxidative stress and hepatopathy in mice. HPLC analysis showed that RGOs were mainly composed of stachyose (61.7%, w/w), followed by 23.7% raffinose and 7.1% sucrose. Administration of RGOs orally daily in mice for 21 days significantly reduced the impact of CCl4 toxicity on the serum markers of liver damage, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total-cholesterol (TC), and triglycerides (TG). RGOs also increased antioxidant levels of hepatic glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC), and ameliorated the elevated hepatic formation of malonaldehyde (MDA) induced by CCl4 in mice, which coincided with the histological alteration. These findings exhibited the potential prospect of RGOs as functional ingredients to prevent ROS-related liver damage.

Dietary supplementation of selenium-enriched tea is known to have an anticancer health benefit. This study was to investigate the inhibitory effect of selenium-containing tea polysaccharides (Se-GTPs) from a new variety of selenium-enriched Ziyang green tea against human MCF-7 breast cancer cells. Se-GTPs dose-dependently exhibited an effective cell growth inhibition with an IC50 of 140.1 μg/mL by inducing MCF-7 cancer cells to undergo G2/M phase arrest and apoptosis. The blockade of cell cycle was associated with an up-regulation of p53 expression, but not CDK2. Se-GTPs clearly triggered the mitochondrial apoptotic pathway, as indicated by an increase in Bax/Bcl-2 ratio and subsequent caspase-3 and caspase-9 activation. It was also found that the generation of intracellular ROS is a critical mediator in Se-GTPs-induced cell growth inhibition. These findings suggest that Se-GTPs may serve as a potential novel dietary agent for human breast cancer chemoprevention.

This study probed the effects of Deshipu stachyose granules (DSG), a novel oligosaccharide preparation (55.3% stachyose, 25.8% raffinose, and 9.7% verbascose), on gut microbiota and constipation in mice. Mice were administered intragastrically without or with DSG (0.42, 0.83, and 2.49 g/kg bw), and feces were collected after 14 days of treatment and subjected to classical microbiological assays. Selective index (SI) and prebiotic index (PI) were incorporated to evaluate the prebiotic effect. DSG at 0.83 g/kg bw scored the highest SI and PI scores, thus supporting a strong prebiotic role. In addition, the impact of DSG (0.42, 0.83, and 1.68 g/kg bw) on defecation function of constipated mice was determined. Ink propulsion rate in the small intestine was significantly improved by DSG treatment. DSG supplementation also distinctly increased the weight and number of black feces within 5 h and evidently shortened the defecating time of first black feces, as compared with the constipation control mice. All of these findings indicate that DSG may promote the growth of beneficial intestinal bacteria and inhibit pathogenic bacteria and also facilitate intestinal peristalsis and fecal excretion, thereby enhancing intestinal health and relieving constipation.

This study was designed to characterize the chemical composition, antioxidant activity and hepatoprotective effect of the polysaccharides from Zizyphus jujube cv. Shaanbeitanzao (ZSP). HPLC analysis showed that ZSP was the heteropolysaccharides with l-arabinose being the main component monosaccharide (50.2%, molar percentage). ZSP displayed strong antioxidant activity in vitro, and the effect was further verified by suppressing CCl4-induced oxidative stress in liver at three tested doses of ZSP (100, 200, and 400 mg/kg BW) in mice. Administration of ZSP (400 mg/kg) significantly (p < 0.01) reduced the activities of CCl4-elevated alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactic dehydrogenase (LDH) in serum, and hepatic malondialdehyde (MDA) level. Mice treated with ZSP showed a better profile of hepatosomatic index (HI) and antioxidant system with normal glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) activities in liver. These results suggest that ZSP exerts an effective protection against CCl4-induced hepatic injury by mediating antioxidative and free radical scavenging activities.Highlights► ZSP was an arabinose-enriched heteropolysaccharide extract. ► A powerful HPLC analysis for PMP-labeled monosaccharides was achieved. ► ZSP possessed high scavenging ability of free radicals. ► ZSP had a significant protective effect against CCl4-induced hepatotoxicity in mice. ► ZSP is a good therapeutic candidate for a valuable hepatoprotection food.

Wolfberry fruit polysaccharides (WFPs) were isolated by hot-water extraction and ethanol precipitation. With HPLC analysis, WFPs were for the first time identified as acidic polysaccharides with galacturonic acid being the main component monosaccharide (24.9%), followed by galactose (21.3%), arabinose (18.5%), and glucose (15.9%), accounting for up to 80.6% of the molar percentage. WFPs exhibited a high antioxidant activity and a dose-dependent antiproliferative effect, with IC50 values of 134.9, 70.1 and 138.4 μg/mL against A549, MCF-7 and LoVo cancer cells after 48 h of incubation as estimated by MTT assay, respectively. Flow cytometry analysis showed that WFPs exerted a stimulatory effect on apoptosis of MCF-7 cells, and induced the cell-cycle arrest at the G0/G1 phase, with the observation of intracellular ROS production and DNA damage. The present study demonstrated that these polysaccharides might have the potential to provide significant natural defence against human cancer.Highlights► Wolfberry fruit polysaccharides (WFPs) exhibited dose-dependent antioxidant and antiproliferative effects. ► MCF-7 cells were more responsive to WFP treatment than A549 and LoVo cells. ► WFPs induced apoptosis and arrest at the G0/G1 phase in MCF-7 cells with DNA damage. ► WFPs are involved in an ROS-dependent apoptosis pathway in MCF-7 cells.

This study was designed to investigate the antioxidant activity, hepatoprotective effect, and phenolic composition of the ethyl acetate fraction (EAF) extracted from Houttuynia cordata tea. EAF was shown to exhibit strong ferric-reducing antioxidant power (FRAP) and scavenging activity against DPPH radical in vitro, and the antioxidant effects were further verified by suppressing CCl4-induced oxidative stress in mouse liver at three tested doses of EAF (250, 500, and 1000 mg/kg bw). Pretreatment with EAF (1000 mg/kg bw) prior to CCl4 administration significantly (p < 0.001) decreased the CCl4-elevated levels of serum AST, ALT, alkaline phosphatase, total bilirubin, and hepatic MDA in mice and prevented the increases in GSH, SOD, and CAT caused by CCl4. HPLC analysis revealed that three predominantly polyphenolic compounds present in EAF were quercitrin (111.7 μg/mg), quercetin (43.8 μg/mg), and hyperoside (29.1 μg/mg). These results combined with liver histopathology indicate that EAF possesses a significant protective effect against acute hepatotoxicity induced by CCl4, which may be due to the strong antioxidant activity of phenolic components.

This study is designed to compare the antioxidant potential of a water-soluble polysaccharide (HCP) with solvent extracts (water, ethanol, ethyl acetate and chloroform) from Houttuynia cordata Thunb. The results showed that polar water extract exhibited the highest reducing power and scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide radical and hydroxyl radical, which were correlated with its high level of biopolymer HCP. Furthermore, the active HCP was identified as an acid hetero-polysaccharide by a rapid HPLC technology within 20 min, and galacturonic acid (29.4%) and galactose (24.0%) were approved as the prominent components of HCP, followed by rhamnose (17.2%), arabinose (13.5%), glucuronic acid (6.8%), glucose (5.3%), xylose (2.1%) and mannose (1.8%) in the molar percentages. This finding suggests that HCP is one of the main active ingredients responsible for antioxidant effect of H. cordata, which might be valuable as a natural antioxidant source applied in both healthy medicine and food industry.

A reversed-phase high-performance liquid chromatographic (HPLC) method is described for the simultaneous determination of aldoses and uronic acids. The separation was carried out on a RP-C18 column (4.6 mm i.d. × 250 mm, 5 μm, Venusil, USA) using precolumn derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) and UV detection at 250 nm, and the 10 PMP derivatives of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose, arabinose and fucose were baseline separated within 40 min. Furthermore, the described method was applied to the quantitative analysis of component monosaccharides in the water-soluble polysaccharides extracted from Gynostemma pentaphyllum Makino tea and the result showed that the tea polysaccharide was a typical heteropolysaccharide and consisted of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose in the molar contents of 16.3, 10.3, 47.1, 5.6, 24.0, 128.4, 25.0, 101.4 and 71.1 μM, respectively. Quantitative recoveries of the component monosaccharides in the tea polysaccharide were in the range of 94.6–108.0% and the RSD values were lower than 4.9%. The results demonstrated that the proposed HPLC method was precise and practical for the analysis of the G. pentaphyllum tea polysaccharide.

To investigate the immunomodulating activity of Radix Angelica sinensis, three different Radix A. sinensis polysaccharide fractions (APFs, namely APF1, APF2 and APF3) were isolated by fractionation using gel filtration and were identified as the immunomodulators of murine peritoneal macrophages. In the present study, it was found that various APFs induced a significant increase in cellular lysosomal enzyme activity, nitric oxide (NO) formation, reactive oxygen species (ROS) production and tumor necrosis factor-α (TNF-α) secretion in macrophages in vitro. Furthermore, APFs dose-dependently stimulated macrophages to produce NO through the up-regulation of inducible NO synthase (iNOS) activity and the maximal effect occurred at a concentration of 500 μg/ml by APF2, followed by APF3 and APF1 in decreasing order. Moreover, the predominant sugars in various APFs were identified as rhamnose, arabonose, glucose, galactose and the ratio of these monosaccharides differed from one polysaccharide fraction to another, which also affected the cellular effector molecule production in macrophages.

The water-soluble crude polysaccharide (CAP) was extracted from Angelica sinensis and was further fractionated by Sephacryl S-400 column chromatography, giving three polysaccharide fractions termed APF1, APF2 and APF3. The obtained results showed that the uronic acid linkages in the polysaccharides were more stable and resistant to thermal hydrolysis than neutral sugar linkages. APF2 exhibited higher thermal stability, followed by APF1, APF3 and CAP in decreasing order. APF1 contained the highest carbohydrate content (80.5%) and the lowest uronic acid content (28.4%) in comparison with APF2 (77.7% and 39.2%) and APF3 (70.1% and 34.6%). Further bioactive investigation showed that intraperitoneal administration of the polysaccharide fractions significantly induced peritoneal macrophage to release nitric oxide (NO), reactive oxygen species (ROS) and enhanced cellular lysosomal enzyme activity (p < .05) and the highest activity was achieved by APF2. Taken together, APF2 had the high thermal stability and immunostimulatory activity and so should be explored as a novel potential immunostimulants.

Water-soluble polysaccharide from Gynostemma pentaphyllum herb tea (PSGP) was isolated by hot-water extraction and ethanol precipitation. The chemical components and preliminary immunomodulating activity of PSGP were investigated both in vitro and in vivo. Capillary zone electrophoresis analysis showed that PSGP was a typical nonstarch heteropolysaccharide, with glucose being the main component monosaccharide (23.2%), followed by galactose (18.9%), arabinose (10.5%), rhamnose (7.7%), galacturonic acid (4.7%), xylose (3.9%), mannose (3.1%), and glucuronic acid (1.2%). PSGP could significantly stimulate peritoneal macrophages to release nitric oxide, reactive oxygen species, and tumor necrosis factor-α in a dose-dependent manner. This immunostimulating activity of PSGP was further demonstrated by its inhibition on the proliferation of human colon carcinoma HT-29 and SW-116 cells incubated with the supernatant of PSGP-stimulated macrophage culture. It is evident that PSGP is a very important ingredient responsible for at least in part the immunomodulating activity of G. pentaphyllum herb tea.

A polyphenol-enriched extract from selenium-enriched Ziyang green tea (ZTP) was selected to evaluate its antitumor effects against human breast cancer MCF-7 cells. In ZTP, (−)-epigallocatechin gallate (28.2%) was identified as the major catechin, followed by (−)-epigallocatechin (5.7%) and (−)-epicatechin gallate (12.6%). ZTP was shown to inhibit MCF-7 cell proliferation (half maximal inhibitory concentration, IC50 = 172.2 μg/mL) by blocking cell-cycle progression at the G0/G1 phase and inducing apoptotic death. Western blotting assay indicated that ZTP induced cell-cycle arrest by upregulation of p53 and reduced the expression of CDK2 in MCF-7 cells. ZTP-caused cell apoptosis was associated with an increase in Bax/Bcl-2 ratio, and activation of caspase-3 and -9. MCF-7 cells treated with ZTP also showed an overproduction of reactive oxygen species, suggesting that reactive oxygen species played an important role in the induction of apoptosis in MCF-7 cells. This is the first report showing that ZTP is a potential novel dietary agent for cancer chemoprevention or chemotherapy.Download full-size image

•RFOs is raffinose family oligosaccharide from Rehmannia glutinosa Libosch.•High l-carnitine diet induce endothelial dysfunction in mice for 10 weeks.•Hepatic injure of mice is induced by long-term ingestion of 3% l-carnitine water.•Intake of RFOs is an effective strategy for prevention of endothelial dysfunction.•RFOs is a potential novel dietary agent for hepatoprotective ingredients.Current research for the first time demonstrated that endothelial dysfunction and hepatic injury in mice were induced by ingestion of 3% l-carnitine water for consecutive 10 weeks. Interestingly, oral administration of dietary raffinose family oligosaccharides (RFOs) at 400 and 800 mg/kg bw significantly reduced the impact of l-carnitine on the serum total cholesterol, triglycerides, high- and low-density lipoproteins, alanine aminotransferase, aspartate amino-transferase, NO, endothelin-1 and C-reactive protein. Furthermore, l-carnitine-induced elevation of hepatic lipid contents and malonaldehyde formation, and the inhibition of SOD and GSH-Px activities in mice were markedly ameliorated by oral administration of RFOs. Moreover, histopathology of H&E and Oil Red O staining of the liver also confirmed the protective effect of RFOs against hepatic steatosis and oxidative injury induced by high l-carnitine diet in mice. These findings for the first time suggest that RFOs may alleviate endothelial dysfunction and liver injury from ingestion of high l-carnitine diet.