Garlic flavonoids alleviate H2 O2 induced oxidative damage in L02 cells and induced apoptosis in HepG2 cells by Bcl-2/Caspase pathway.

Liver damage is a common liver disorder, which could induce liver cancer. Oral antioxidant is one of the effective treatments to prevent and alleviate liver damage. In this study, three flavonoids namely myricetin, isoquercitrin, and isorhamnetin were isolated and identified from Laba garlic. The isolated compounds were investigated on the protective effects against H2 O2 -induced oxidative damages in hepatic L02 cells and apoptosis inducing mechanism in hepatic cancer cells HepG2 by using MTT assay, flow cytometry and western blotting analysis. Myricetin, isoquercitrin, and isorhamnetin showed proliferation inhibition on HepG2 cells with IC50 value of 44.32 ± 0.213 µM, 49.68 ± 0.192 µM, and 54.32 ± 0.176 µM, respectively. While they showed low toxicity on normal cell lines L02. They could significantly alleviate the oxidative damage towards L02 cells (P < 0.05), via inhibiting the morphological changes in mitochondria and upholding the integrity of mitochondrial structure and function. The fluorescence intensity of L02 cells pre-treated with myricetin, isoquercitrin, and isorhamnetin (100 µM) was 89.23 ± 1.26%, 89.35 ± 1.43% and 88.97 ± 0.79%, respectively. Moreover, the flavonoids could induce apoptosis in HepG2 cells via Bcl-2/Caspase pathways, where it could up-regulate the expression of Bax and down-regulate the expression of Bcl-2, Bcl-xL, pro-Caspase-3, and pro-Caspase-9 proteins in a dose dependent manner. Overall, the results suggested that the flavonoids from Laba garlic might be a promising candidate for the treatment of various liver disorders. PRACTICAL APPLICATION: Flavonoids from Laba garlic showed selective toxicity towards HepG2 cells in comparison to L02 cells via regulating Bcl-2/caspase pathway. Additionally, the isolated flavonoids expressively barred the oxidative damage induced by H2 O2 in L02 cells. These results suggested that the flavonoids from laba garlic could be a promising agent towards the development of functional foods.

Probing the structure-antioxidant activity relationships of four cinnamic acids porous starch esters.

For investigation of antioxidant capacity relationship, four cinnamic acids (CNAs), including cinnamic (CA), ferulic (FA), p-coumaric (p-CA) and sinapic (SA) acids, were selected to modify porous starch (PS) with different degrees of substitution by esterification, respectively. The ester linkage of CNAs modified PS was confirmed by 1H NMR, 13C solid-state NMR and FT-IR. The porous structure was maintained after esterification. Three in vitro antioxidant assays were applied to measure antioxidant capacities. The order of antioxidant capacity was SA@PS > FA@PS > p-CA@PS > CA@PS, due to the presence of phenolic hydroxyl groups with hydrogen donating abilities. Besides electron-donating group on ortho or para positions the benzene ring further enhances the hydrogen donating ability and the stability of hydroxyl radical. This study not only investigated the antioxidant mechanism of CNA modified starch derivatives but probed the way for synthesis of biodegradable antioxidant materials for the food industries.

Ultrafiltration isolation, physicochemical characterization, and antidiabetic activities analysis of polysaccharides from green tea, oolong tea, and black tea.

Both fermentation degree and preparation method of polysaccharides could influence the bioactivity of tea polysaccharides. The aim of this study was to compare the physicochemical characterization and biological activities of the polysaccharides isolated by ultrafiltration method from three kinds of tea (green tea, oolong tea, and black tea). The bioactivities of tea polysaccharide fractions were compared from four aspects, including antioxidant activities, antiglycation activities, α-glucosidase inhibitory capability, and hypoglycemic effects on L6 myotubes. Results showed that six polysaccharides (GTPS1, GTPS2, OTPS1, OTPS2, BTPS1, and BTPS2) had different contents of neutral sugar and uronic acid, and they showed different morphologies. Six polysaccharides were composed of the seven monosaccharides with different molar ratios. BTPS1 exhibited the highest DPPH scavenging activity and hydroxyl radical scavenging activity (P < 0.05), and BTPS1 also showed the strongest antiglycation inhibitory effects (P < 0.05). BTPS1 and BTPS2 showed strong inhibitory capacity on α-glucosidase and hypoglycemic effects in L6 skeletal muscle cells. The result suggested that the degree of fermentation of tea could improve their bioactivities (BTPS > OTPS >GTPS), and TPS1 with smaller molecular weight distribution showed higher bioactivities than TPS2. This study can provide a scientific foundation for the application of tea polysaccharides and related functional products.

Characterization, antioxidant activities, and inhibition on α-glucosidase activity of corn silk polysaccharides obtained by different extraction methods.

The polysaccharides (CSPw, CSPc, CSPa, and CSPu) were prepared by hot water extraction, acid-assisted extraction, alkaline-assisted extraction, and ultrasound-assisted extraction from corn silk, respectively. High performance gel permeation chromatography (HPGPC), fourier-transform infrared (FT-IR) spectroscopy, and scanning electron microscopy (SEM) results indicated that the extraction methods had an obvious impact on the molecular weight, structure, and morphology of the CSPs. Among the four polysaccharides, CSPu showed the highest inhibitory α-glucosidase activity, which might be related to its smaller molecular weight. Furthermore, kinetics analyses revealed that CSPu had significant inhibition of α-glucosidase in a non-reversible and competitive manner. Fluorescence quenching analysis illustrated that the interaction mechanism of CSPu and α-glucosidase was claimed as a static quenching mechanism. Isothermal titration calorimetry (ITC) analysis showed that the main driving forces for the interaction of CSPu with α-glucosidase was hydrogen bonding and the binding interactions of them occurred spontaneously.

Virtual Screening and Optimization of Novel mTOR Inhibitors for Radiosensitization of Hepatocellular Carcinoma.

BACKGROUND: Radiotherapy has an ameliorative effect on a wide variety of tumors, but hepatocellular carcinoma (HCC) is insensitive to this treatment. Overactivated mammalian target of rapamycin (mTOR) plays an important part in the resistance of HCC to radiotherapy; thus, mTOR inhibitors have potential as novel radiosensitizers to enhance the efficacy of radiotherapy for HCC. METHODS: A lead compound was found based on pharmacophore modeling and molecular docking, and optimized according to the differences between the ATP-binding pockets of mTOR and PI3K. The radiosensitizing effect of the optimized compound (2a) was confirmed by colony formation assays and DNA double-strand break assays in vitro. The discovery and preclinical characteristics of this compound are described. RESULTS: The key amino acid residues in mTOR were identified, and a precise virtual screening model was constructed. Compound 2a, with a 4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidine scaffold, exhibited promising potency against mTOR (mTOR IC50=7.1 nmol/L (nM)) with 126-fold selectivity over PI3Kα. Moreover, 2a significantly enhanced the sensitivity of HCC to radiotherapy in vitro in a dose-dependent manner. CONCLUSION: A new class of selective mTOR inhibitors was developed and their radiosensitization effects were confirmed. This study also provides a basis for developing mTOR-specific inhibitors for use as radiosensitizers for HCC radiotherapy.

Antioxidant, α-amylase and α-glucosidase activity of various solvent fractions of I. obliquus and the preventive role of active fraction against H2 O2 induced damage in hepatic L02 cells as fungisome.

Inonotus obliquus is a traditional mushroom well known for its therapeutic value. In this study, various solvent fractions of I. obliquus were preliminarily screened for their antioxidant, α-amylase and α-glucosidase inhibition properties. To improve the drug delivery, the active fraction (ethyl acetate fraction) of I. obliquus was synthesized into fungisome (ethyl acetate phophotidyl choline complex, EAPC) and its physical parameters were assessed using Fourier transform infrared spectroscopy (FTIR), High performance liquid chromatography (HPLC), Scanning electron microscope (SEM), and ς potential analysis. Then normal human hepatic L02 cells was used to evaluate the cytotoxicity of EAPC. The results showed that EA fraction possesses significant free radical scavenging, α-amylase and α-glucosidase inhibition properties. FTIR, SEM, and HPLC analysis confirmed the fungisome formation. The particle size of EAPC was 102.80 ± 0.42 nm and the ς potential was -54.30 ± 0.61 mV. The percentage of drug entrapment efficiency was 97.13% and the drug release rates of EAPC in simulated gastric fluid and simulated intestinal fluid were 75.04 ± 0.29% and 93.03 ± 0.36%, respectively. EAPC was nontoxic to L02 cells, however it could selectively fight against the H2 O2 induced oxidative damage in L02 cells. This is the first study to provide scientific information to utilize the active fraction of I. obliquus as fungisome. PRACTICAL APPLICATIONS: Inonotus obliquus (IO) is a traditional medicinal fungus. The extracts of IO have obvious antioxidant and hypoglycemic activities. Ethyl acetate (EA) fraction of IO was encapsulated in liposomes to form EAPC. EAPC has a sustained-release effect. It has nontoxic to L02 cells and could protect L02 cells from oxidative damage caused by hydrogen peroxide. This study could provide new ideas for the treatment of diabetes.

Antioxidant and antihypertensive effects of garlic protein and its hydrolysates and the related mechanism.

Garlic protein (GP) was enzymatically hydrolyzed using pepsin and trypsin followed by the evaluation of antioxidant and angiotensin-converting enzyme (ACE) inhibitory activities of GP and its hydrolysates. The antihypertensive effects of GP and its hydrolysates were determined in vivo. The results showed that GP and its hydrolysates namely GPH-P (pepsin) and GPH-T (trypsin) possessed appreciable antioxidant and ACE inhibitory activities. The ACE inhibitory activity of GP, GPH-T, and GPH-P was in consistent with their antioxidant activities. GP and its hydrolysates offered significant protective effects against H2 O2 -induced oxidative damage (p < .05). In addition, the administration of GP, GPH-T, and GPH-P reduced the blood pressure in hypertensive rats. The mechanism might be to reduce blood pressure by inhibiting the activity of ACE, reducing the formation of ACEII, and protecting the activity of bradykinin. This study suggested that GP might be utilized as a promising functional food as antioxidant and antihypertensive agents. PRACTICAL APPLICATIONS: Garlic (Allium sativum L.) is one of the oldest cultivated plants that belongs to the Liliaceae family and it has been used in cooking and medicinal applications. Large quantities of garlic residuals were produced with the development of the garlic essential oil industry. These residuals are highly rich in proteins and they can be used for the preparation of protein hydrolysates. Generally, hydrolysates are considered as a promising food supplement due to the enrichment of amino acids present in it. In this study, garlic proteins (GPs) and its hydrolysates retain effective antioxidant effects. They were found to reduce  the blood pressure and prevent oxidative stress induced by H2 O2 . The information from this study could be used to develop a new nutritional supplement from GP and its hydrolysates to treat hypertension as well as prevent oxidative damage.

Effectiveness and safety of light vegetarian diet on functional constipation with gastrointestinal damp-heat pattern: An exploratory study protocol for randomized controlled trial.

INTRODUCTION: Functional constipation (FC) is one of the common gastrointestinal disorders that affects people of almost every age. Persistent FC significantly affects quality of life and well-being along with economic burden on patients as well as health care system. Therapeutic efficacy of currently used treatment strategies becomes limited shortly after their discontinuation as constipation occurs again as a result of inappropriate dietary habits. Previous studies have revealed that light vegetarian diet (LVD) can significantly improve both typical and atypical subtypes of major traditional Chinese medicine (TCM) FC syndrome such as gastrointestinal damp-heat syndrome. This protocol aims at exploratorily investigating effectiveness and safety of LVD following a rigorous clinical trial. METHODS AND DESIGN: Total 92 patients in each of the 2 subtypes will be recruited in China-Japan Friendship Hospital for participating in this prospective, placebo-controlled, randomized trial and exploratory study. The patients in each subtype will be randomly divided into 4 groups according to 1:1:1:1 ratio with allocation concealment, which are drug + diet group, drug group, placebo + diet group and placebo group. Patients in the group with diet intervention will be required to strictly follow the LVD. The study will continue for a period of 28 days, including a drug or placebo supervised intervention and a 14th-day telephone follow-up. During the intervention, patients will be required to record a designed diary for controlling the diet quality (DQ) and analyzing the defecation. The study will focus investigation of complete spontaneous bowel movements (CSBM) per week as its primary outcome and constipation-related symptom rating scale (CSS), TCM syndrome scale (TCMSS), 48-hour gastrointestinal transit time (48-hour GITT), high resolution anorectal manometry (HRAM) and fecal flora detection (FFD) will be included in secondary outcomes. Furthermore, the study will also determine safety, DQ and compliance indicators. ETHICS AND DISSEMINATION: This study has been approved by China-Japan Friendship Hospital clinical research ethics committee (No. 2017-46-1). A SPIRIT checklist is available for this protocol. TRIAL REGISTRATION NUMBER: ChiCTR1800019686 in Chinese Clinical Trial Registry (WHO ICTRP member).

Identification of Ellagitannins in the Unripe Fruit of Rubus Chingii Hu and Evaluation of its Potential Antidiabetic Activity.

As a functional food, the unripe fruits of Rubus chingii Hu have been widely used in China for thousands of years. Twenty-five major ellagitannins (ETs) were identified from the unripe fruits, and a novel ellagitannin, chingiitannin A (1), together with four other known ETs (2-5) were isolated and identified by HPLC-QTOF-MS/MS and 2D-NMR. Chingiitannin A showed the highest α-glucosidase and α-amylase inhibitory activities (IC50 2.89 and 4.52 µM, respectively), which occurred in a reversible and noncompetitive manner. Static quenching was indicated in a fluorescence quenching assay. Molecular docking results revealed that chingiitannin A interacted with the enzymes mainly by hydrogen bonding and was bound in the allosteric site. Chingiitannin A was nontoxic, and it increased the glucose uptake in L6 myotubes. The results suggested that the unripe fruits of Rubus chingii Hu are rich sources of ETs, and chingiitannin A might be a good candidate for functional foods or antidiabetic drugs.

Identification and antimicrobial activity evaluation of three peptides from laba garlic and the related mechanism.

Laba garlic is a traditional Chinese processed garlic (Allium sativum L.) with multiple health benefits. This study was aimed at isolating and identifying three antimicrobial peptide fractions from laba garlic using Sephadex chromatography, pre-HPLC and LC-MS/MS and investigating the potential antimicrobial mechanism. The sequences of the three peptides were demonstrated to be Tyr-Asn-His-Asn-Phe (YNHNF, F3-3-a), Trp-Pro-Thr-Ser-Phe-Thr (WPTSFT, F3-3-b) and Ala-Val-Asp-Arg-Ala-Val (AVDRAV, F3-3-c), respectively. F3-3-c showed the strongest anti-microbial effects among these three fractions and it could inhibit the mycelial growth of E. coli and S. aureus in vitro with a minimum inhibitory concentration (MIC) of 100 µM and exhibited negligible hemolytic activity. F3-3-c showed amphipathic properties with the presence of α-helix (25.8 ± 0.56%), ß-strands (19.7 ± 0.45%), turns (21.2 ± 0.78%) and unordered conformation (33.3 ± 1.09%). Treatment of F3-3-c resulted in a change in the morphology of the hyphae and the disruption of membrane integrity as shown by SEM and TEM analyses. These results suggested that the laba garlic peptide F3-3-c might be a promising candidate for antibiotics and related functional foods.

Effects of N-trans-feruloyltyramine isolated from laba garlic on antioxidant, cytotoxic activities and H2O2-induced oxidative damage in HepG2 and L02 cells.

Laba garlic is a kind of processed garlic products, it is the traditional Chinese food with a long history. In this study, the antitumor, antioxidant and cytotoxic properties of the blue pigment (BP) from Laba garlic were investigated. N-trans-feruloyltyramine (FLA) was isolated and identified from BP. The protective effects of FLA against H2O2-induced oxidative damages in L02 cells were also assessed. The apoptotic effects of FLA were detected by using flow cytometry analysis. Results showed that the tumor growth was significantly suppressed by BP (P<0.05). BP and FLA exhibited remarkable antioxidant activities. L02 cells pretreatment with FLA could significantly fight against the oxidative damage induced by H2O2, inhibit the morphological changes of mitochondria and maintain the integrity of mitochondria. FLA showed proliferation inhibition on HepG2 cells with IC50 value of 194 ±â€¯0.894 µM. After treatment of FLA (320 µM), the results of MTT assay on HepG2 and L02 cells indicated that FLA had selective cytotoxic effects. It suggested a new way of prevention and treatment of tumors and FLA might be a promising candidate in cancer therapy and functional foods.

Structural characterization of corn silk polysaccharides and its effect in H2O2 induced oxidative damage in L6 skeletal muscle cells.

To evaluate the antioxidant activity of polysaccharides from corn silk in H2O2 induced oxidative damage in L6 skeletal muscle cells. A polysaccharide (CSP2) was isolated from corn silk. Structure analysis of CSP2 with 1D and 2D NMR indicated that CSP2 was composed of (1→6)-linked α-d-glucose. The substituent of the α-d-glucose is composed of (1→3)-linked α-l-arabinose, (1→4)-linked ß-d-galactose and (1→3,5)-linked ß-d-mannose, with ß-d-xylose and α-l-rhamnose as terminal unit. CSP2 had the potential to scavenge DPPH and hydroxyl radical and inhibit hemolysis in vitro. CSP2 was found to suppress oxidant stress by improving the enzyme activities of SOD, CAT, and GPX. Rodamine 123 staining results showed that the pretreatment of CSP2 prevented changes in the mitochondrial membrane potential and increased the fluorescence intensity in L6 cells in the presence of H2O2. These results suggested that CSP2 could be utilized as a potential antioxidant supplement to prevent oxidative stress.

Interaction characterization of preheated soy protein isolate with cyanidin-3-O-glucoside and their effects on the stability of black soybean seed coat anthocyanins extracts.

The interactions of soy protein isolate with cyanidin-3-O-glucoside were investigated to study the protective effect of protein on anthocyanin's stability by UV-Vis spectrophotometry, Fourier transform infrared spectroscopy, circular dichroism and fluorescence spectroscopy. Preheat treatment and binding of cyanidin-3-O-glucoside effectively changed the secondary structure of soy protein isolate, with a decrease in α-helix, random coil structure and an increase in ß-sheet and ß-turn. The soy protein isolate preheated at 121 °C exhibited a strong binding affinity towards cyanidin-3-O-glucoside with strong Ks of 147.40 × 104 M-1 and also effectively increased the thermal and oxidation stabilities of black soybean seed coat extract via decreasing the degradation rate by 67% and 23%, respectively. Soy protein isolate interacted with cyanidin-3-O-glucoside mainly through hydrophobic interactions and static quenching process. Altogether, the results suggested that preheated soy protein isolate-cyanidin-3-O-glucoside interaction could effectively protect anthocyanins' stability through strong binding affinity influenced by the systematic alterations in the secondary structure.

Physicochemical properties of polysaccharides from Lentinus edodes under high pressure cooking treatment and its enhanced anticancer effects.

This study was to investigate the physicochemical properties and anticancer effects of polysaccharides from Lentinus edodes extracted under high pressure cooking treatment (HPLPS) in vitro and in vivo. The extraction efficiency was improved. The main molecular weight of HPLPS was about 540 and about 227 kDa. And the inhibitory effects on HepG2 and HeLa cells of HPLPS were significantly increased (p < 0.05). The in vivo anticancer effect on H22 tumor bearing mice model was evaluated. The tumor growth inhibitory rate of HPLPS-H was 67.66%. The activities of ALT and AST were decreased. The activities of SOD, CAT, GSH-Px were notably increased. The expressions of IL-2 and TNF-α were increased while the expression of VEGF was decreased. These results suggested that high pressure-assisted extracted polysaccharides from Lentinus edodes might be effectively used for the treatment of hepatocellular carcinoma through its antioxidant and immunomodulatory effects.

Protective effect of the total flavonoids from Apocynum venetum L. on carbon tetrachloride-induced hepatotoxicity in vitro and in vivo.

Apocynum venetum L., belonging to the family Apocynaceae, is a popular medicinal plant, which is commonly used in the treatment of hypertension, neurasthenia, and hepatitis in China. In the present study, the total flavonoids (TFs) were prepared from the leaves of A. venetum, and its protective effects on carbon tetrachloride (CCl4)-induced hepatotoxicity in a cultured HepG2 cell line and in mice were investigated. Cell exposed to 0.4% CCl4 (v/v) for 6 h led to a significant decrease in cell viability, increased LDH leakage, and intracellular reactive oxygen species (ROS). CCl4 also induced cell marked apoptosis, which was accompanied by the loss of mitochondrial membrane potential (MMP). Pretreatment with TFs at concentrations of 25, 50, and 100 µg/mL effectively relieved CCl4-induced cellular damage in a dose-dependent manner. In vivo, TFs (100, 200, and 400 mg/kg BW) were administered via gavage daily for 14 days before CCl4 treatment. The high serum ALT and AST levels induced by CCl4 were dose-dependently suppressed by pretreatment of TFs (200 and 400 mg/kg BW). Histological analysis also supported the results obtained from serum assays. Furthermore, TFs could prevent CCl4-caused oxidative damage by decreasing the MDA formation and increasing antioxidant enzymes (CAT, SOD, GSH-Px) activities in liver tissues. In summary, both in vitro and in vivo data suggest that TFs, prepared from A. venetum, showed a remarkable hepatoprotective and antioxidant activity against CCl4-induced liver damage.

Hypoglycemic and hypolipidemic effects of anthocyanins extract from black soybean seed coat in high fat diet and streptozotocin-induced diabetic mice.

Black soybean seed coat extract (BSSCE) is a rich source of anthocyanins with multiple health effects. This study was aimed at investigating the composition and hypoglycemic and hypolipidemic effects of BSSCE in vitro and in a high-fat diet and streptozotocin (STZ)-induced diabetic mice. The anthocyanins of BSSCE were identified as cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, and peonidin-3-O-glucoside by HPLC-MS. Results demonstrated that BSSCE exhibited strong inhibitory activities for α-amylase, potent inhibition activity against lipid accumulation in HepG2 cells and protection effect on H2O2-induced oxidative stress-damaged HepG2 cells. The food and water intake, body weight loss, blood glucose and insulin level of BSSCE treatment group were found to be significantly reduced when compared with those of diabetic mice group (p < 0.05). The fasting blood glucose level and insulin level of the BSSCE 400 mg kg-1 group mice significantly decreased by 47.97% and 46.49%, respectively. The oral glucose tolerance and activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) notably improved (p < 0.05). BSSCE could also ameliorate the atherogenic dyslipidaemia of diabetic mice by remarkably decreasing the content of total cholesterol (T-CHO), total triglyceride (TG), and non-esterified fatty acid (NEFA) and increasing the content of high-density lipoprotein cholesterol (HDL-c) (p < 0.05). BSSCE could protect against liver, kidney and pancreas damages in diabetic mice. This study suggested that cyanidin-3-O-glucoside contributed to BSSCE-induced hypoglycemia and hypolipidemia effects in type 2 diabetes mellitus (T2DM), and BSSCE might be a promising functional food or medicine for T2DM treatment.

Anti-diabetic effects of Inonotus obliquus polysaccharides-chromium (III) complex in type 2 diabetic mice and its sub-acute toxicity evaluation in normal mice.

Polysaccharides are important bioactive ingredients from Inonotus obliquus. This study aimed to synthesize and characterize a novel I. obliquus polysaccharides-chromium (III) complex (UIOPC) and investigate the anti-diabetic effects in streptozotocin (STZ) induced type 2 diabetes mellitus (T2DM) mice and sub-acute toxicity in normal mice. The molecular weight of UIOPC was about 11.5 × 10 4 Da with the chromium content was 13.01% and the chromium was linked with polysaccharides through coordination bond. After treatment of UIOPC for four weeks, the body weight, fasting blood glucose (FBG) levels, plasma insulin levels of the diabetic mice were significantly reduced when compared with those of the diabetic mice (p < 0.05). The results on serum profiles and antioxidant enzymes activities revealed that UIOPC had a positive effect on hypoglycemic and antioxidant ability. Histopathology results showed that UIOPC could effectively alleviate the STZ-lesioned tissues in diabetic mice. Furthermore, high dose administration of UIOPC had no obviously influence on serum profiles levels and antioxidant ability of the normal mice and the organ tissues maintained organized and integrity in the sub-acute toxicity study. These results suggested that UIOPC might be a good candidate for the functional food or pharmaceuticals in the treatment of T2DM.