Extract of Euryale ferox Salisb exerts antidepressant effects and regulates autophagy through the adenosine monophosphate-activated protein kinase-UNC-51-like kinase 1 pathway.

Positive regulation of autophagy by Euryale ferox Salisb (ES) leads to antidepressant effects. This study focused on the potential antidepressant mechanisms induced by the petroleum ether fraction of ES (ES-PE) in the chronically unpredictable mild stress (CUMS) mouse model. Deficits in the sucrose preference test, open field test, tail suspension test, and forced swim test were observed following CUMS, and were reversed following ES-PE administration. Contrary to the expression of mammalian target of rapamycin, autophagy was decreased after establishment of the CUMS model. We also observed trends for downregulation of adenosine mononphosphate-activated protein kinase (AMPK) and mammalian autophagy-initiating kinase (ULK1), which were differentially affected by ES-PE. HT22 cells and Compound C, an inhibitor of AMPK, were used to verify the results in mice. Gas chromatography-mass spectrometry analysis revealed high level of vitamin E acetate in ES-PE. Taken together, our data indicate that ES-PE activated autophagy by regulating the AMPK pathway. © 2018 IUBMB Life, 70(4):300-309, 2018.

Study on the Structure of Ginseng Glycopeptides with Anti-Inflammatory and Analgesic Activity.

Panax ginseng is well known for its medicinal functions. As a class of important compound of ginseng, ginsenoside is widely studied around the world. In addition, ginseng glycopeptides also showed good biological activity, but researches in this field are rarely reported. In this study, ginseng glycopeptides (Gg) were first prepared from Panax ginseng by reflux extracted with 85% ethanol and the following purification with Sephadex G-15 column. Then, the inflammatory pain models induced by carrageenan and the rat pain models induced by Faure Marin were established for research on mechanism of analgesic activities. It is showed that Gg had an obvious inhibiting effect on inflammation and a significant reduction on the Malondialdehyde (MDA) of inflammatory foot tissue. And there were significant differences between moderate to high dose of Gg and model group in Interleukin 1ß (IL-1ß), Interleukin 2 (IL-2), Interleukin 4 (IL-4), Tumor necrosis factor α (TNF-α) and Histamine. The two models can be preliminarily determined that the analgesic effect of Gg may be peripheral, which mechanism may be related to the dynamic balance between proinflammatory cytokines (TNF-α, IL-1ß) and anti-inflammatory cytokines (IL-2, IL-4, and Interleukin 10 (IL-10)). A series of methods were used to study Gg in physical-chemical properties and linking mode of glycoside. The high-resolution mass spectrometry was used for identification of the structure of Gg. Moreover, the structure of 20 major Gg were investigated and identified. The structural analysis of Gg was benefit for the next study on structure-activity relationship.

Effect of the molecular mass of tremella polysaccharides on accelerated recovery from cyclophosphamide-induced leucopenia in rats.

The body of tremella were decocted with water, and hydrolyzed with 0.1 mol/L hydrochloric acid for different times, giving tremella polysaccharides with six molecular mass values. The structures of all the tremella polysaccharides had non-reducing terminals of ß-D-pyranglucuronide, the backbone was composed of (1 → 3)-linked ß-D-manno-pyranoside, and the side chain composed of (1 → 6)-linked ß-D-xylopyranoside was attached to the C(2) of the backbone mannopyranoside. Immunomodulatory effect studies indicated that tremella polysaccharides increased the counts of leukocytes in the peripheral blood which were significantly lowered by cyclophosphamide, and the lower the molecular mass of the tremella polysaccharide, the better this effect was.

Animal welfare information affects consumers' hedonic and emotional responses towards milk.

This research aimed to explore the effects of animal welfare information on consumers' hedonic and emotional responses towards milk. Two studies were conducted. For Study 1, participants (N = 101) were asked to fill out a questionnaire on attitudes towards animal welfare, in which a variety of factors including raising methods, quality of life, emotions, quality of the product, nutrition, price, and environment, were tested. For Study 2, participants (N = 63) tasted a milk sample (2% fat, standardized and homogenized) in two different conditions: [1] blind (without any previous information), and [2] informed (with information stating that the milk was obtained from a farm with an animal welfare system in place). For Study 1, participants with higher milk consumption per week showed a higher agreement with positive animal welfare statements. For Study 2, the overall liking for the milk in the informed condition was significantly higher than that of the milk in the blind condition (7.4 vs. 6.8, using a 9-point hedonic scale). Participants had higher penalizations for the milk in the blind condition as they suggested that the milk's flavor, sweetness, aroma, and mouthfeel were not enough in the product. This research showed that animal welfare can be an important extrinsic factor in the consumers' hedonic and emotional responses towards milk. These findings can be useful for understanding consumers' behaviors towards animal welfare.

Structure-activity relationship analysis of Panax ginseng glycoproteins with cytoprotective effects using LC-MS/MS and bioinformatics.

Panax ginseng glycoproteins (PGG) has been shown biological activity, but researches in this field are rarely reported. In this paper, PGG were prepared by reflux and then purified with macroporous resin column. Further separation and purification of PGG using high performance liquid chromatography (HPLC) and two major components (PGG-1, PGG-2) were obtained. The molecular weights were calculated by gel permeation chromatography (GPC), and the results are 1.5 KDa and 8.2 KDa respectively. The MTT assay was used to study the cytoprotective effects of PGG, the results exhibited that PGG had significant effect (P < 0.01), and showed an obvious dose-effect relationship. Anti-apoptosis experiment results showed that PGG and PGG-2 can inhibit Aß-induced apoptosis in SH-SY5Y cells (P < 0.05), and PGG-2 displayed better activity. The structures of N- and O-glycan were determined by combination of LC-MS/MS and methylation analysis. The computed parameters of PGG determined by MS including the theoretical isoelectric point (pI), instability index, aliphatic index and grand average of hydropathicity (GRAVY) were summarized systematically. The distinct differences between two parts would affect the behavior of PGG in vivo. The results of activity test and bioinformatics analysis would guide the study of PGG in pharmacokinetics and mechanism.

LC-MS/MS analysis of partial structure of Panax ginseng protein and its distribution in vivo.

Protein from Panax ginseng can improve learning, memory, and analgesia. Here, we investigated a fluorescence labeling method that can be used to determine the in vivo distribution of P. ginseng protein (PGP). High-performance liquid chromatography (HPLC) was used to define the amino acid composition and molecular weight of PGP; LC-MS/MS was used to identify the PGP structure, which was fluorescently-labeled using a fluorescein isothiocyanate (FITC) probe. The connection form of the PGP fluorescent marker (PGP-FITC) was identified by ultraviolet and infrared spectrophotometry. The in vivo distribution of PGP was observed by fluorescence imaging, and tissue content was determined. Results showed that PGP was enriched in the brain and that vascular epithelial cells showed specific uptake. We provide an experimental method to label and identify the in vivo distribution of PGP, which forms the basis for future studies to determine whether PGP can penetrate the blood-brain barrier (BBB) and elucidate the transport mechanism.

Metabonomics Study of Ginseng Glycoproteins on Improving Sleep Quality in Mice.

The changes of brain metabolism in mice after injection of ginseng glycoproteins (GPr) were analyzed by gas chromatography mass spectrometry- (GC/MS-) based metabolomics platform. The relationship between sedative and hypnotic effects of ginseng glycoproteins and brain metabolism was discussed. Referring to pentobarbital sodium subthreshold test, we randomly divided 20 mice into two groups: control and ginseng glycoproteins group. The mice from the control group were treated with normal saline by i.p and GPr group were treated with 60 mg/kg of GPr by i.p. The results indicated that GPr could significantly improve the sleep quality of mice. Through multivariate statistical analysis, we found that there were 23 differential metabolites in whole brain tissues between the control group and the GPr group. The pathway analysis exhibited that GPr may be involved in the regulation of the pathway including purine metabolism, nicotinate and nicotinamide metabolism, glycine, serine and threonine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, and steroid hormone biosynthesis. This work is helpful to understand the biochemical mechanism of GPr on promoting sleep and lay a foundation for further development of drugs for insomnia.

The antidepressant effects of apigenin are associated with the promotion of autophagy via the mTOR/AMPK/ULK1 pathway.

The present study aimed to investigate whether apigenin elicits antidepressant effects in depressant­like mice via the regulation of autophagy. The depressant­like behaviors were established in a chronic restraint stress model. Male BALB/c mice were subjected to restraint stress for 6 h/day for a period of 21 days, and deficits in sucrose preference, tail suspension and forced swim tests were confirmed to be improved following oral apigenin. To investigate the underlining mechanisms, the hippocampal levels of p62 and microtubule­associated protein light chain 3­II/I (LC3­II/I) were measured using western blot analysis. The expression levels of LC3­II/I and p62 indicated that the significantly inhibited autophagy level induced by chronic restraint stress can be increased following apigenin treatment. Similar to the level of autophagy, the expression levels of adenosine monophosphate­activated protein kinase (AMPK) and Unc­51 like autophagy activating kinase­1 were downregulated after chronic restraint stress stimulation and, subsequently upregulated following treatment with apigenin. Conversely, the levels of mammalian target of rapamycin (mTOR) were increased in chronic restraint stress mice and inhibited by apigenin. Collectively, the present findings indicated that apigenin potentially promotes autophagy via the AMPK/mTOR pathway and induces antidepressive effects in chronic restraint stress mice.

Crocin induces anti-ischemia in middle cerebral artery occlusion rats and inhibits autophagy by regulating the mammalian target of rapamycin.

Crocin, an active compound found in Gardenia jasminoides Ellis, has been shown to possess neuron-protective properties, but its potential mechanisms of action still remain poorly understood. In this study, the anti-ischemic effect and underlying mechanism of action of crocin were investigated in male rats with right middle cerebral artery occlusion/reperfusion. Computed tomography and magnetic resonance imaging were used to evaluate the area of infarction 24 h after reperfusion. Neurological scores were employed to evaluate nerve injury. Direct 2,3,5-triphenyltetrazolium chloride staining was used to calculate the infarct ratio 120 h after reperfusion. Finally, HT22 cells and Western blot were used to study the underlying mechanisms. Crocin showed a decreased infarct volume and neurological score in vivo, while the expression of LC3-II/I and AMP-activated protein kinase was remarkably down-regulated with increased levels of p62 and mammalian target of rapamycin (mTOR) expression. However, rapamycin significantly inhibited mTOR, which can impact the anti-ischemic effect of crocin in vitro. These results suggest that crocin may elicit an anti-ischemic effect probably through the mTOR pathway.

In vivo and in vitro neuroprotective effects of Panax ginseng glycoproteins.

The root of Panax ginseng C. A. Mey (Araliaceae) has medicinal value in complex system of Traditional Chinese medicines for its use in improving cognitive function. A glycoproteins named PGL-1 was extracted from ginseng which subjected to through a macroporous resin, hollow-fiber ultrafiltration and dialyzed. The glycoproteins has a molecular weight in the range from 0.4 to 4.4kDa, with an average molecular mass of 1.6kDa. HPLC analysis revealed that the compositions of glycoproteins included fucose, mannose, rhamnose, glucose, galacturonic acid, N-acetylglucosamine and N-acetylgalactosamine. Glycan of PGL-1 has a backbone of →4)-Rha-(1→, →4)-Fuc -(1→, →6)-Gal-(1→, →4)-GalA-(1→, →4)-GlcNAc-(1→ and →4)-GalNAc-(1→,and (→3,6)-Man-(1→) was distributed in branches. The (1→)-Fuc, (1→)-Glc and (1→)-GlcNAc or (1→)-GalNAc were regarded as a terminal residue. The Morris water maze test revealed that the PGL-1 can effectively alleviate the memory impairment symptoms of rats induced by Aß25-35. All dose groups showed significant activity of protective effect on apoptosis SH-SY5Y induced by Aß25-35, and obviously inhibited the S phase arrest. Compared with Aß25-35 treatment alone, a significant reduction in NO concentration and NOS activity was detected in cells co-administered with glycoproteins. Thus, glycoproteins derived from ginseng might be a promising anti-AD reagent.

The antidepressant effects of ɑ-tocopherol are related to activation of autophagy via the AMPK/mTOR pathway.

The purpose of the present study was to investigate whether ɑ-tocopherol exhibited neuro-protective effects in chronic unpredictable mild stress (CUMS) mice through the regulation of autophagy. Deficits in behavioural tests, including a sucrose preference test, open field test, forced swim test, and tail suspension test, were ameliorated following ɑ-tocopherol administration. To study the potential mechanism, western blots were performed on both prefrontal cortex and hippocampus samples. Similar to the degree of autophagy, the activities of adenosine monophosphate-activated protein kinase (AMPK) and Unci-51 like autophagy activating kinase-1 (ULK1) were decreased after CUMS stimulation. In addition, we also found increased activity of the mammalian target of rapamycin (mTOR), which was significantly affected following administration of ɑ-tocopherol, as well as its three downstream pathways. Taken together, our study found that ɑ-tocopherol might potentially promote autophagy to induce anti-depressive responses in CUMS mice though the AMPK/mTOR pathway.

Analgesic effects of glycoproteins from Panax ginseng root in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Panax ginseng C.A. Mey has various beneficial pharmacological effects. The present study aimed to evaluate the analgesic activities of glycoproteins from the root of Panax ginseng C.A. Mey in mice. MATERIALS AND METHODS: Glycoproteins were isolated and purified from the root of Panax ginseng C.A. Mey. Physicochemical properties and molecular mass were determined by chemical assay and HPLC. Acetic acid-induced writhing and hot-plate tests were employed to study the analgesic effect of glycoproteins and compared with that of aspirin or morphine. The locomotor activity was tested in mice by using actophometer. RESULTS: Four glycoproteins were obtained. The glycoproteins which protein content was the highest (73.04%) displayed dose-dependent analgesic effect. In writhing test, the glycoproteins significantly inhibited writhes (P<0.001) at the dose of 20 mg/kg by intraperitoneal injection. In hot-plate test, only at the dose of 20 mg/kg prolong the hot-plate latency (P<0.05, at 30 min). In the locomotor activity test, the glycoproteins were significant decrease of motility counts at the dose of 20 and 40 mg/kg. CONCLUSION: These findings collectively indicate that the glycoproteins from the root of Panax ginseng C.A. Mey exhibited significant analgesic activities and the proteins were the active site, providing evidence for its pharmacal use.

Studies on the characteristic and activity of low-molecular fragments from zymosan.

Zymosan was hydrolysed with HCl and fractionated by ultrafiltration and dialysis to obtain water-soluble fragments A, B and C. Physical and chemical analyses showed that these fractions are composed primarily of glucose and have molecular weights of 8 kDa, 5 kDa and 2 kDa, respectively. A glycosidic linkage analysis indicated that they are mainly composed of ß-1,3-glucans. Fragment A, which has the highest molecular weight, contains approximately 30% ß-1,6-linked glucans, but fragment C is almost entirely composed of linear ß-1,3-glucan chains. The anti-chronic atrophic gastritis activity experiments showed that fragment A has significant activity, the activity of zymosan is quite low and the activities of fragments B and C are in between those of fragment A and zymosan.