Quantitative Mitochondrial Proteomics Study on Protective Mechanism of Grape Seed Proanthocyanidin Extracts Against Ischemia/Reperfusion Heart Injury in Rat

Cardiac ischemia/reperfusion（I/R） injury is a critical condition,often associated with high morbidity and mortality.The cardioprotective effect of grape seed proanthocyanidin extracts（GSPE） against oxidant injury during I/R has been described in previous studies.However,the underlying molecular mechanisms have not been fully elucidated.This study investigated the effect of GSPE on reperfusion arrhythmias especially ventricular tachycardia（VT） and ventricular fibrillation（VF）,the lactic acid accumulation and the ultrastructure of ischemic cardiomyocytes as well as the global changes of mitochondria proteins in in vivo rat heart model against I/R injury.GSPE significantly reduced the incidence of VF and VT,lessened the lactic acid accumulation and attenuated the ultrastructure damage.Twenty differential proteins related to cardiac protection were revealed by isobaric tag for relative and absolute quantitation（iTRAQ） profiling.These proteins were mainly involved in energy metabolism.Besides,monoamine oxidase A（MAOA） was also identified.The differential expression of several proteins was validated by Western blot.Our study offered important information on the mechanism of GSPE treatment in ischemic heart disease.

Grape Seed Proanthocyanidin Extract Alleviates Arsenic-induced Oxidative Reproductive Toxicity in Male Mice

Objective To determine the ability of grape seed proanthocyanidin extract （GSPE） in alleviating arsenic-induced reproductive toxicity. Methods Sixty male Kunming mice received the following treatments by gavage： normal saline solution （control）; arsenic trioxide （ATO; 4 mg/kg）; GSPE （400 mg/kg）; ATO＋GSPE （100 mg/kg）; ATO＋GSPE （200 mg/kg） and ATO＋GSPE （400 mg/kg）. Thereafter, the mice were sacrificed and weighed, and the testis was examined for pathological changes. Nuclear factor （erythroid-derived 2）-like 2 （Nrf2）, heme oxygenase 1 （HO1）, glutathione S-transferase （GST）, NAD（P）H dehydrogenase, and quinone 1 [NQO1） expression in the testis was detected by real-time PCR. Superoxide dismutase （SOD）, glutathione （GSH）, total antioxidative capability （T-AOC）, malondialdehyde （MDA）, 8-hydroxydeoxyguanosine （8-OHdG）, and reproductive indexes were analyzed. Results ATO-treated mice showed a significantly decreased sperm count and testis somatic index and activity levels of SOD, GSH, and T-AOC than control group. Compared to the ATO-treated group, ATO ＋GSPE group showed recovery of the measured parameters. Mice treated with ATO＋high-dose GSPE showed the highest level of mRNA expression of Nrf2, HO, NO.O1, and GST. Conclusion GSPE alleviates oxidative stress damage in mouse testis by activating Nrf2 signaling, thus counteracting arsenic-induced reproductive toxicity.

Inhibitory Effects of Grape Seed Proanthocyanidin Extract on Selenite-induced Cataract Formation and Possible Mechanism

This study investigated the inhibitory effect of grape seed proanthocyanidin extract(GSPE) on selenite-induced cataract formation in rats and the possible mechanism.Eighty 8-day-old Sprague-Dawley rats were divided randomly into 5 groups:control group,model group,three GSPE groups(low dose,medium dose and high dose).Control group received subcutaneous injection of physiological saline.Model group was given subcutaneous injection of sodium selenite(20 μmol/kg body weight) on the postpartum day 10,and once every other day for consecutive three times thereafter.GSPE treated groups were respectively administered GSPE at doses of 50,100,and 200 mg/kg body weight intragastrically 2 days prior to the selenite injection(that was,on the postpartum day 8),and once daily for fourteen consecutive days thereafter.The opacity of lenses was observed,graded and photographed under the slit lamp microscopy and the maximal diameter of the nuclear cataract plaques was measured.The lenses were analyzed for superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-PX),malondialdehyde(MDA),calcium(Ca 2+),nitric oxide(NO) and anti-hydroxyl radical ability(anti-OH).The histomorphology of lenses was observed with HE staining under a light microscope.The levels of calpainⅡ,and iNOS protein and mRNA expression in lenses were detected by using immunohistochemistry and real-time quantitative RT-PCR.The results showed subcutaneous injection of sodium selenite led to severe nuclear cataract in model group,and the achievement ratio of model group was 100%.As compared with model group,the degree of lenses opacity and the maximal diameter of nuclear cataract plaques were significantly reduced in GSPE-treated groups.Moreover,we observed selenite treatment caused a significant decrease in the activities of antioxidative enzymes(SOD,CAT,GSH-PX) and anti-OH ability,accompanied by a significant increase in the levels of MDA,NO,Ca 2+ as well as iNOS,and calpainⅡ protein and mRNA expression.Administration of GSPE could dose-dependently preserve the activities of these antioxidative enzymes and anti-OH ability,accompanied by a significant reduction in the levels of MDA,NO,Ca 2+ as well as iNOS,and calpainⅡ protein and mRNA expression.These results suggested that GSPE markedly prevented selenite-induced cataract formation probably by suppressing the generation of lipid peroxidation and free radicals as well as the activation of iNOS,and calpainⅡ in the lenses.

Orbital decompression surgery and horse chestnut seed extract improved superior orbital vein blood flow in patients with thyroid-associated ophthalmopathy

AIM： To evaluate the efficacy and safety of orbital decomposition （OD） surgery in combination with horse chestnut seed extract （HCSE）, as compared to OD atone, in patients with thyroid-associated ophthalmopathy （TAO）. METHODS： Sixty-two orbits from 62 TAO patients were randomly assigned to OD or OD＋HCSE at 1：1 ratio （31 received OD alone, 31 received OD ＋HCSE）. Forty-two orbits from 21 healthy subjects were used as controls. Complete ophthalmic examination and color Doppler flow imaging （CDFI） were performed before surgery and 3mo post-surgery on all 62 orbits from the TAO patients. CDFI were also performed on the 42 control orbits, The effect of OD ＋HCSE and OD alone on TAO orbits was compared on several endpoints, including superior ophthalmic vein blood flow （SOVBF） parameters, subjective assessment, soft tissue involvement, lid retraction, diplopia, eye movement restriction, degree of exophthalmos, and intraocular pressure. The control orbits were used as reference for the SOVBF parameters. RESULTS： OD surgery with or without HCSE improved SOVBF, symptoms and soft tissue involvement, decreased degree of exophthalmos and intraocular pressure in orbits of TAO patients. The OD ＋HCSE combination led to significantly better improvement of SOVBF than OD alone. The differences between the reductions of SOVBF in the two groups are 1.26 cmls in max-volecity and 0.52 cm/s in min-voiecity （P〈0.0001）. CONCLUSION： SOVBF is significantly reduced in the orbits affected with TAO, indicating that congestion may be an important factor contributing to TAO pathogenesis. OD surgery improves the SOVBF, and combination of HCSE medication and OD surgery further improved venous return than OD surgery alone.

Grape Seed Proanthocyanidin Extract Ameliorates Streptozotocin-induced Cognitive and Synaptic Plasticity Deficits by Inhibiting Oxidative Stress and Preserving AKT and ERK Activities

Progressive memory loss and cognitive impairment are the main clinical manifestations of Alzheimer’s disease(AD).Currently,there is no effective drug available for the treatment of AD.Previous studies have demonstrated that the cognitive impairment of AD is associated with oxidative stress and the inhibition of AKT and ERK phosphorylation.Grape seed proanthocyanidin extract(GSPE)has been shown to have strong antioxidant effect and can protect the nervous system from oxidative stress damage.This study aimed to investigate the protective effect of GSPE on the cognitive and synaptic impairments of AD using a sporadic AD rat model induced by intracerebroventricular(ICV)injection of streptozotocin(STZ)(ICV-STZ).Rats were treated with GSPE(50,100,or 200 mg/kg every day)by intragastrical(ig.)administration for continuous 7 weeks,and ICV-STZ(3 mg/kg)was performed on the first day and third day of week 5.Learning and memory abilities were assessed by the Morris water maze(MWM)test at week 8.After behavioral test,hippocampal long-term potentiation(LTP)was recorded,and the levels of malondialdehyde(MDA),superoxide dismutases(SOD),glutathione(GSH)and the protein expression of AKT and ERK were measured in the hippocampus and cerebral cortex of rats.Our study revealed that ICV-STZ significantly impaired the working learning ability and hippocampal LTP of rats,significantly increased the levels of MDA,and decreased the activity of SOD and GSH in the hippocampus and cerebral cortex.In contrast,GSPE treatment prevented the impairment of cognitive function and hippocampal LTP induced by ICV-STZ,decreased the level of MDA,and increased the level of SOD and GSH.Furthermore,Western blot results showed that GSPE treatment could prevent the loss of AKT and ERK activities in the hippocampus and cerebral cortex induced by ICV-STZ.Our findings demonstrate that GSPE treatment could ameliorate the impairment of cognitive ability and hippocampal synaptic plasticity in a rat model of sporadic AD by inhibiting oxidative stress and preserving AKT and ERK activities.Therefore,GSPE may be an effective agent for the treatment of cognitive deficits associated with sporadic AD.

Effects of grape seed extract on meat color and premature browning of meat patties in high-oxygen packaging

This study investigated the effects of grape seed extract(GSE)on fresh and cooked meat color and premature browning(PMB)in ground meat patties(85% beef and 15% pork back fat)packaged under high-oxygen modified atmospheres(HiOx-MAP).The GSE was added to patties at concentrations of 0,0.10,0.25,0.50 and 0.75 g kg^(-1).This study evaluated the surface color,pH,lipid oxidation,and total viable counts(TVC)of raw patties,and the internal color and pH of patties cooked to a temperature of 66 or 71℃ over 10-day storage at 4℃.Compared with the control(0 g kg^(-1) GSE),GSE improved the color stability(P<0.05)and significantly inhibited the lipid and myoglobin oxidation of raw patties from day 5 to 10,but GSE had no effect(P>0.05)on TVC.Patties containing 0.50 and 0.75 g kg^(-1) GSE cooked to 66℃ exhibited greater(P<0.05)interior redness than the control and reduced the PMB of cooked patties in the late storage stage.These results suggested that 0.50 and 0.75 g kg^(-1) GSE can improve fresh meat color and minimize PMB of HiOx-MAP patties.

Add-on therapy of herbal formulation rich in standardized fenugreek seed extract in type 2 diabetes mellitus patients with insulin therapy: An efficacy and safety study

Objective: To assess the safety and efficacy of herbal formulation rich in standardized fenugreek seed extract(IND-2) add-on therapy in type 2 diabetes mellitus(T2DM) patients who were on insulin treatment in prospective, single arm, open-label, uncontrolled, multicentre trial.Methods: T2DM patients(n=30) with aged 18-80 years who were stabilized on insulin treatment with fasting blood sugar(FBS) level between 100-140 mg/dL received IND-2 capsules(700 mg, thrice a day) for 16 weeks.The primary endpoints were an assessment of FBS at week 2, 4, 6, 8, 12 and 16.Secondary end-points include post-prandial blood sugar level, glycosylated Hb(HbA1c), reduction in the dose of insulin and number of hypoglycemic attacks, and improvement in lipid profile at various weeks.Safety and adverse events(AEs) were also assessed during the study.Results: Study was completed in twenty T2DM patients, and there was no significant reduction in FBS and post-prandial blood sugar level after addon therapy of IND-2.However, add-on therapy of IND-2 significantly reduced(P<0.01) the HbA1c values, requirements of insulin and hypoglycemic events as compared with baseline.Total cholesterol, high-density lipoproteins-cholesterol, and low-density lipoproteincholesterol levels were significantly increased(P<0.01) after IND-2 add-on therapy.Body weight and safety outcomes did not differ significantly in IND-2 add-on therapy group at week 16.Additionally, add-on therapy of IND-2 did not produce any serious adverse events.Conclusions: The results of present investigation suggest that add-on therapy of IND-2 with insulin in T2DM patients improves glycaemic control through a decrease in levels of HbA1c and number of insulin doses needed per day without an increase in body weight and risk of hypoglycemia.Thus, IND-2 may provide a safe and well-tolerated add-on therapy option for the management of T2DM.

Grape Seed Procyanidin Extract Reduces Arsenic- Induced Renal Inflammatory Injury in Male Mice

The aim of the present study is to evaluate the ability and mechanism by which grape seed procyanidin extract （GSPE） relieves arsenic trioxide （As2O3）-induced renal inflammatory injury. Therefore, male Kunming mice were treated with As2O3 and/or GSPE by gavage for 5 weeks. Mice were then sacrificed and inflammatory cytokines of kidneys were examined by ELISA, whereas the expression levels of molecules involved in the nuclear factor （NF）-KB signaling pathway were evaluated by both qRT-PCR and Western blot. Our results indicate that GSPE prevents As2O3-mediated renal inflammatory injury by inhibiting activation of the NF-KB signaling pathway and inflammatory cytokine production, while promoting expression of anti-inflammatory cytokines.

Vitamin C, Grape Seed Extract and Citrus Bioflavonoids Protect the Skin against Photoaging: A Review

The skin is a major protective organ of the body. It is constantly exposed to the environment and is very resilient. But exposure to ultraviolet (UV) rays from the sun results in the production of reactive oxygen species (ROS) and subsequent inflammatory responses that can overwhelm the innate protective mechanisms of the skin. This results in damage and premature aging. Strategies to mitigate this premature photoaging might include avoidance of sunlight. However, some sunlight exposure is beneficial to health. One notable example of this is the production of vitamin D. A more practical approach to preventing adverse effects of UV light in the skin is antioxidant supplementation. Dietary antioxidants may help control ROS propagation following UV light exposure. To further evaluate the utility of antioxidants in protecting the skin, in vitro, in vivo and human studies of three well known dietary antioxidants are reviewed and discussed. The data clearly demonstrate that vitamin C, grape seed extract and citrus bioflavonoids have the potential to reduce the damaging effects of excess sun exposure via antioxidant, anti-inflammatory and immunomodulating mechanisms. As such, regular ingestion of dietary antioxidants appears to be a useful strategy for protecting the skin against photoaging.

Grape Seed Extract and the Fetal Ductus Arteriosus: A Potential Danger of a Common Herbal Supplement

A female at 28 weeks gestation was referred to pediatric cardiology for a fetal arrhythmia. The echocardiogram revealed premature constriction of the fetal ductus arteriosus. Her work up was unremarkable except for her use of an herbal supplement, grape seed extract, which is advertised as a potent anti-inflammatory medication, and has biochemical properties similar to other medications that have been shown to cause premature ductal constriction. The use of herbal remedies increases each year. Although the public is inundated with radio, television, and internet advertisements for these products, little unbiased information regarding the possible dangers of toxicity or adverse reactions exists. As physicians, we need to be aware of these products, and counsel our patients accordingly.

Uromedic^(■) Pumpkin Seed Derived Δ7-Sterols, Extract and Oil Inhibit 5α-Reductases and Bind to Androgen Receptor in Vitro

Dihydrotestosterone (DHT) is implicated in the development of benign prostate hyperplasia (BPH). We investigated if Uromedic? pumpkin (variety of Cucurbita pepo L. convar. citrullinina GREB. var. styriaca GREB) seed soft extract (active ingredients of GRANUFINK? Prosta forte 500 mg), seed oil and isolated Δ7-sterols could inhibit the conversion of [1,2,6,7-3H(N)]-testosterone to DHT by 5α-reductases. Also, we tested competition with [3H]-DHT for binding to the androgen receptor (AR). Pumpkin seed oil and pumpkin seed soft extract were identified as moderately active inhibitors of 5α-R1 and 5α-R2, with almost similar inhibitory capacities (IC50 5 mg/ml for 5α-R1 and about IC50 = 6 mg/ml for 5α-R2). The isolated Δ7-sterols were more active inhibitors (IC50 = 0.3 mg/ml for 5α-R1, IC50 = 1.0 mg/ml for 5α-R2). All three test compounds bound to the AR dose-dependently, with strong binding by Δ7-sterols (IC50 = 0.2 mg/ml) and weaker binding by pumpkin seed oil (IC50 = 0.4 mg/ml) and pumpkin seed soft extract (IC50 = 1.1 mg/ml). We propose that inhibition of 5α-reductases and competitive binding to the AR are mechanisms of action, by which the Uromedic? pumpkin seed derived test compounds, most specifically Δ7-sterols, counteract DHT and thereby exert clinically positive effects on the prostate, as well as bladder-strengthening effects.

Effects of Citrullus colocynthis Seed Aqueous Extracts upon Sodium Transport across A6 Kidney Cell Monolayers

Background:Various extracts from Citrullus colocynthis seeds were recently examined for their possible favorable effects on glucose homeostasis,other metabolic variables,and pancreatic islet size in streptozotocin-induced diabetic rats.Objective:Considering the effectiveness of insulin upon target cells as another process susceptible to be influenced by plant extracts,the present study is used to compare the effects of either crude or defatted aqueous extracts from C.colocynthis seeds upon Na+transport in A6 kidney cells.Method:The intensity of electrical currents,taken as the potential difference divided by the epithelial resistance was measured in monolayers of A6 kidney cells exposed at the basolateral side to either crude untreated aqueous(EI)or defatted aqueous(EII)extracts,as well as their fractions EI1 and EII1 containing components with high molecular weight(>5 kDa),and EI2 and EII2 containing components with low molecular weight(<5 kDa).The effect of insulin was to assess the functional integrity of the A6 monolayer.Results:All extracts caused a rapid and sustained increase in current intensity.The defatted aqueous extract(EII)was more efficient than the non-defatted aqueous extract(EI)in terms of increasing the intensity of current in the A6 monolayer.The components of the defatted aqueous extract responsible for such a greater efficiency were located in the fraction containing components with low molecular weight and acted in a time-related progressive fashion upon Na+transport.Conclusion:These data suggest that colocynth seed extracts mainly containing components with low molecular weight increased the current intensity in A6 cells,thus suggesting that colocynth may affect Na+transport in kidney cells.

A novel approach of proteomics to study the mechanism of action of grape seed proanthocyanidin extracts on diabetic retinopathy in rats

Background Diabetic retinopathy （DR） is a leading cause of visual impairment and blindness among the people of occupational age. To prevent the progress of retina injury, effective therapies directed toward the key molecular target are required. Grape seed proanthocyanidin extracts （GSPE） have been reported to be effective in treating diabetic complications, while little is discussed about the functional protein changes. Methods We used streptozotocin （STZ） to induce diabetes in rats. GSPE （250 mg/kg body weight per day） were administrated to diabetic rats for 24 weeks. Serum glucose, glycated hemoglobin and advanced glycation end products （AGEs） were determined. Consequently, 2-D difference gel electrophoresis and mass spectrometry were used to investigate retina protein profiles among control, STZ-induced diabetic rats, and GSPE treated diabetic rats. Results GSPE significantly reduced the AGEs of diabetic rats （P 〈0.05）. Moreover, GSPE significantly suppressed the vascular lesions of central regions, decreased capillary enlargements and neovascularization, similar to those of the control rats under light microscope. Eighteen proteins were found either up-regulated or down-regulated in the retina of STZ-induced diabetic rats. And seven proteins in the retina of diabetic rats were found to be back-regulated to normal levels after GSPE therapy. These back-regulated proteins are involved in many important biological processes such as heat shock, ubiquitin-proteasome system, cell proliferation, cell growth and glucose metabolism. Conclusions These findings might promote a better understanding for the mechanism of DR, and provide novel targets for evaluating the effects of GSPE therapy.

Effects of Manual, Cultural, Botanical and Chemical Treatments of Termite Control in Hamelmalo Agricultural College Area

Termites are the most serious pests of field and horticultural crops, forests, and wooden household furniture. In Hamelmalo Agricultural College (HAC) the infestation of termite is very high resulting in great destruction of crop plants and wooden office and dormitory furniture. The devastating attack of termites should be managed by using best and ecofriendly management method. The purpose of the study was to compare the effectiveness of manual destruction of mounds and killing of termite queen and king, chemical chlorpyrifos, seed and leaves extract of neem and Lantana (as separate experiment) and smoke on termite control. Termite mounds were selected randomly inside HAC compound. The materials used were hand hoe, spade, fork, water, 20 L jar and protective clothes. The treatments were replicated three times. The botanical treatments were prepared at 2 L of highly concentrated extracts per 20 L of water each. Chlorpyrifos was applied at 20 ml per 20 L of water. Dried woody plants were used for smoke treatment. Careful digging was done to avoid king escape and queen rupture and they were killed by burning. Among all, the mechanical destruction and killing of king and queen and chlorpyrifos resulted in a complete control of the termite population. Except in the mounds treated by chlorpyrifos, the activity of termite population was very active and they closed the opened galleries immediately after treatment even though there were dead termite castes in all treatments. Living termite castes were counted by taking a medium size spade of broken mound pieces. The highest count was recorded from mounds treated by smoke. After two weeks the queen and king in every treatment mound were cheeked and killed for those who were alive. Except by the chlorpyrifos and manual destruction of mound (king and queen were killed before) all the royal families were alive and killed. Controlling of termite population in the field (outside their mound) is not possible due to the hidden foraging activity of termites, environmental safety from chemicals and the high egg laying potential of the queen. Finding the best alternative to control from their source mound for the mound building termites resulted in effective control of the population by manual destruction of mounds and killing of queen and king and chlorpyrifos. By the side effect of chlorpyrifos to untargeted organisms and the time consuming and laborious method of manual destruction of mounds, selection to the best from these two control measure is almost the same.

Callogenesis and Antibacterial Activity of Balanites aegyptiaca

B. aegyptiaca, it is a species of economic and cultural importance in various countries, with diverse uses that include: medicinal, charcoal, pesticides and forage and in vitro callus production is important to have many applications in both basic and industrial research on this specie. For the induction of callus, B. aegyptica seed cotyledons were surface sterilized with 90% ethanol for 1 minute and cultivated in MS media supplemented with 2,4-D, BAP and NAA. Both the callus and seed were collected and dried in an oven at 40°C - 45°C. Cotyledon’s seed and callus were grounded into the powdered form using mortar and pestle and stored at room temperature for further use. Five grams (5 g) each of the powder were mixed with 50 ml of the solvents: methanol and n-hexane (1:10) w/v, agitated vigorously and kept on an orbital shaker at 150 rpm for 24 h, then filtered. The extracts of the plant sample were evaluated in agar dilution method which was used to determine the MIC and MBC of the extracts. The auxin NAA in low concentrations (0.5 mg/L) in the presence of a dose of 0.5 mg/L of the cytokinin BAP induced 100% callus formation. The 50 and 100 mg/ml methanolic extracts were more effective than the n-hexane extracts for both the gram-positive and gram-negative bacteria. By callus extracts under 100 and 50 mg/ml reveals that methanolic extracts of callus had the highest zone of inhibition. An effective protocol for callus induction has been developed that can use for germplasm conservation or for genetic engineering. Evidence from the present study revealed both extracts possess strong broad-spectrum antibacterial effect. Therefore, methanolic extract of seed kernel callus of B. aegyptiaca can be utilized as a new source of broad spectrum antibacterial drugs for effective control of bacteria related diseases.

Grape Seed Extract Attenuates Demyelination in Experimental Autoimmune Encephalomyelitis Mice by Inhibiting Inflammatory Response of Immune Cells

Objective: To examine the anti-inflammatory effect of grape seed extract(GSE) in animal and cellular models and explore its mechanism of action. Methods: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis(MS)-experimental autoimmune encephalomyelitis(EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17and the inflammatory factors such as tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6, IL-12, IL-17and interferon-γ(IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay(ELISA), immunofluorescence staining and Western blot, respectively. Results: GSE reduced the secretion of TNF-α, IL-1β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide(P<0.01), inhibited the secretion of TNF-α, IL-1β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days(P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors(P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4(TLR4) and Rho-associated kinase(ROCKⅡ, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors(P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45+CD11b+and CD45+CD4+cells, and weakened the differentiation of Th1 and Th17(P<0.05). Moreover, it reduced the secretion of inflammatory factors(P<0.01), and prevented the activation of microglia(P<0.05). Conclusion: GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.

Transcriptome profile and its partly verification of human hepatocellular carcinoma cells exposed to Yuzhizi(Fructus Akebiae Quinatae) seed extract

OBJECTIVE: To investigate the efficacy of Yuzhizi(Fructus Akebiae Quinatae) seed extract(FAQSE) on inhibiting the proliferation of hepatocellular carcinoma(HCC) cells in vitro and to explore the anti-HCC action mechanism of FAQSE. METHODS: Human HCC Hep G2 and Huh7 cells were used to investigate the anti-HCC effect of FAQSE in vitro. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) method was used to measure cell viability. Affymetrix microarray was adopted to detect the expression of transcriptome. The differentially expressed genes(DEGs) of each cell line were identified. For coDEGs of both cell lines, the gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway were enriched using the Database for Annotation, Visualization and Integrated Discovery(DAVID), and the network analysis of protein-protein interaction(PPI) was mapped using the Retrieval of Interacting Genes/Proteins(STRING) and Cytoscape software. Some important genes in the PPI network of coDEGs were selected to verify by quantitative real-time reverse transcription-polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay. RESULTS: FAQSE decreased the viability of Hep G2 and Huh7 cells. There were 211 co-upregulated and 86 codownregualted genes in both cell lines after FAQSE treatment. The enriched GO terms of co-upregulated DEGs were primarily involved cell-cell adhesion, viral process, transcription initiation from RNA polymerase II promoter, positive regulation of transcription from RNA polymerase II promoter and actin cytoskeleton organization. The GO terms of co-downregulated DEGs were mainly enriched in the processes of SRP-dependent cotranslational protein targeting to membrane, viral transcription, nuclear-transcribed m RNA catabolic process, nonsense-mediated decay, translational initiation and r RNA processing. Main KEGG pathways of co-upregulated DEGs were endocytosis, glutathione metabolism, protein processing in endoplasmic reticulum, synaptic vesicle cycle and lysosome. The major KEGG pathways of co-downregulated DEGs were ribosome, biosynthesis of amino acids, arginine and proline metabolism, systemic lupus erythematosus and complement and coagulation cascades. The top 10 coDEGs with high hub nodes in STRING analysis were ribosomal protein S27a, transferrin, ribosomal protein S20, ribosomal protein L9, protein phosphatase 2 regulatory subunit B alpha, transthyretin, thioredoxin reductase 1, ribosomal protein L3, ribophorin I and ribosomal protein L24. Alpha-fetoprotein(AFP) was also co-downregulated and contained in the PPI network. The m RNA and protein expression of most verified genes was consistent with the results of co-DEGs analysis. And the AFP level was significantly reduced after FAQSE treatment. CONCLUSIONS: A series of genes and pathways of Hep G2 and Huh7 cells were changed after FAQSE treatment, which might be the targets of FAQSE against HCC and worthy of further study. AFP might be important one of them.

Buffalo stirred yoghurt fortified with grape seed extract:New insights into its functional properties

Grape seed extract(GSE)is considered one of the major byproducts,which are rich in phenolic content and possess several health-promoting activities including antibacterial,anticancer,and antioxidant.The current research was devoted to improve the bioactive properties of stirred yoghurt through supplementation with GSE at three concentrations(0.1,0.25,and 0.5 g/100 g yoghurt).The obtained stirred yoghurts were analyzed for determining their composition,pH,syneresis,viscosity,and sensory evaluation.Furthermore,some functional properties including antioxidant,antibacterial,anticancer activities were estimated.Significant increments(p≤0.05)were observed in total solids,ash contents,pH,water holding capacity,and viscosity values,especially when 0.5%GSE was added.Moreover,adding 0.5%GSE showed higher(p≤0.05)in vitro antioxidant,antibacterial,and anticancer activities.Interestingly,the improved bioactive properties resulting from adding GSE,even at the higher concentration,were not accompanied by any change in sensory properties.In particular,the color descriptor was mostly preferred by panelists when 0.5%GSE was added.The assessors did not observe any off-flavors in any of the tested stirred yoghurt samples.Therefore,the functional properties of stirred yoghurt could be improved by adding GSE preparation without affecting its organoleptic properties.