Quercetin protects against hyperglycemia-induced retinopathy in Sprague Dawley rats by regulating the gut-retina axis and nuclear factor erythroid-2-related factor 2 pathway.

Hyperglycemia-related retinopathy is a disease with a high blindness rate. Recent reports indicate that many flavonol compounds have the potential to prevent the occurrence of disease in the retina by regulating the gut-retina axis. Here, we hypothesized that quercetin could alleviate the symptoms of retinopathy. To clarify the mechanism, Sprague Dawley rats were fed a high-fat diet containing quercetin for 12 weeks and injected with streptozotocin in the ninth week. Additionally, neomycin and ampicillin were used to establish a pseudo-sterile rat model. Afterward, changes in the retina were investigated by using electroretinogram and optical coherence tomography. Blood and tissue samples were collected and biochemical components were analyzed. The extent of intestinal injury was determined via hematoxylin-eosin staining. Microbial community structure was analyzed by using 16S ribosomal RNA sequencing. Finally, the expression of genes was analyzed using real-time polymerase chain reaction. The results showed that quercetin reduced the decline in electroretinography amplitude and outer nuclear layer thickness, increased the activities of antioxidant enzymes, decreased the contents of proinflammatory factors and blood glucose, enhanced the concentration of insulin, and inhibited intestinal dysbiosis and improved gut morphology. Importantly, the underexpression of nuclear factor erythroid-2 related factor 2 in the retina was reversed by quercetin. However, trend changes were no longer significant in most of the indicators after antibiotic treatment. In summary, quercetin has therapeutic effects on retinopathy by regulating the gut-retina axis and nuclear factor erythroid-2 related factor 2 pathway, and the presence of gut microbiota helps quercetin exert its effects on the retina.

Nervonic acid reduces the cognitive and neurological disturbances induced by combined doses of D-galactose/AlCl3 in mice.

Nervonic acid (NA) is a kind of ultra-long-chain monounsaturated fatty acid, which can repair nerve cell damage caused by oxidative stress. Alzheimer's disease (AD) is a nervous system disease and often accompanied by the decline of learning and memory capacity. In this study, the combined dose of D-galactose/AlCl3 was used to establish a mouse model of AD. Meanwhile, the mice were treated with different doses of NA (10.95 and 43.93 mg/kg). The results showed that NA delayed the decline of locomotion and learning ability caused by D-galactose/AlCl3, increased the activity of total superoxide dismutase, catalase, glutathione peroxidase, and reduced the content of malondialdehyde in vivo. Besides, NA reduced the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), aspartate aminotransferase, alanine aminotransferase, increased the levels of 5-hydroxytryptamine, dopamine, γ-aminobutyric acid, alleviated the cell morphology damage induced by D-galactose/AlCl3 in hippocampus and liver tissue. Furthermore, the intervention of NA upregulated the expression levels of PI3K, AKT, and mTOR genes and downregulated the expression levels of TNF-α, IL-6, and IL-1ß genes. Therefore, we speculate the intervention of NA could be an effective way in improving cognitive impairment through the activation of PI3K signaling pathway. These results suggest that NA has the potential to be developed as antioxidant drug for the prevention and early therapy of AD.

Hyperoside inhibits pancreatic lipase activity in vitro and reduces fat accumulation in vivo.

Hyperoside, the main component of many anti-obesity plants, might exhibit a lipase inhibition effect to reduce fat accumulation. The anti-obesity effect of hyperoside was investigated by studying its inhibitory effect and mechanism on pancreatic lipase in vitro and evaluating its ability to reduce lipid accumulation in vivo. Hyperoside is a mixed-type inhibitor of lipase with an IC50 of 0.67 ± 0.02 mmol L-in vitro. Hyperoside changed the secondary conformation of lipase, increased the α-helix content, and changed the microenvironment of lipase through static quenching. The interaction between hyperoside and lipase results from a strong binding spontaneous exothermic reaction, mainly through hydrogen bonding, van der Waals force and electrostatic force. Hyperoside protected hepatic lipid accumulation and adipose tissue hypertrophy and reduced the expression of inflammatory factors in high-fat diet-induced rats. Moreover, hyperoside had a good inhibitory effect on lipase activity in serum and increased fecal fat excretion, thereby reducing lipid absorption in vivo. This study provides theoretical support for the research and development of hyperoside in fat-reducing functional foods.

Spoof Trace Disentanglement for Generic Face Anti-Spoofing.

Prior studies show that the key to face anti-spoofing lies in the subtle image patterns, termed "spoof trace," e.g., color distortion, 3D mask edge, and Moiré pattern. Spoof detection rooted on those spoof traces can improve not only the model's generalization but also the interpretability. Yet, it is a challenging task due to the diversity of spoof attacks and the lack of ground truth for spoof traces. In this work, we propose a novel adversarial learning framework to explicitly estimate the spoof related patterns for face anti-spoofing. Inspired by the physical process, spoof faces are disentangled into spoof traces and the live counterparts in two steps: additive step and inpainting step. This two-step modeling can effectively narrow down the searching space for adversarial learning of spoof trace. Based on the trace modeling, the disentangled spoof traces can be utilized to reversely construct new spoof faces, which is used as data augmentation to effectively tackle long-tail spoof types. In addition, we apply frequency-based image decomposition in both the input and disentangled traces to better reflect the low-level vision cues. Our approach demonstrates superior spoof detection performance on 3 testing scenarios: known attacks, unknown attacks, and open-set attacks. Meanwhile, it provides a visually-convincing estimation of the spoof traces. Source code and pre-trained models will be publicly available upon publication.

Phlorizin alleviates cholinergic memory impairment and regulates gut microbiota in d-galactose induced mice.

We explored the effect of phlorizin against cholinergic memory impairment and dysbacteriosis in D-galactose induced ICR mice. The control (CON) group, D-galactose model (DGM) group, and three groups (DG-PL, DG-PM, DG-PH) treated with phlorizin at 0.01%, 0.02%, and 0.04% (w/w) in diets were raised for 12 weeks. Supplementing with phlorizin reversed the loss of organ coefficient and body weight caused by D-galactose. The functional abilities of phlorizin on hippocampal-dependent spatial learning and memory, anti-oxidation, anti-inflammation were also observed. Meanwhile, phlorizin intervention upregulated the gene expression of Nrf2, GSH-PX, SOD1, decreased the gene expression of NF-κB, TLR-4, TNF-α, and IL-1ß in the hippocampus, while enhanced the gene expression of JAM-A, Mucin2, Occludin in the caecum. Furthermore, a neurotransmitter of acetylcholine (ACh) was enhanced, while acetylcholinesterase (AChE) activity was inhibited by phlorizin administration. Moreover, phlorizin administration increased short-chain fatty acids (SCFAs) content, and reduced lipopolysaccharides (LPS) levels, which may relate to the rebuilding of gut microbiota homeostasis. Treatment with phlorizin may be an effective intervention for alleviating cognitive decline and gut microbiota dysbiosis.

Phlorizin exerts potent effects against aging induced by D-galactose in mice and PC12 cells.

Phlorizin is the main active ingredient of apple peel and has potential utilization value. Some recent studies have suggested that phlorizin may have antioxidant capacity and protect the liver. The injection of a low dose of d-galactose can cause some changes that resemble accelerated aging in mice. This study explored the protective effects of phlorizin on d-galactose-induced mice and PC12 cells. In this study, ICR mice were divided into a normal group (NOR), a d-galactose model group (d-gal) and phlorizin treatment groups (100 mg kg-1, 200 mg kg-1 and 400 mg kg-1). In addition to the NOR group, four other groups were injected with d-galactose (120 mg kg-1) for 12 weeks. The results showed that phlorizin reduced the decline of strength, coordination and spatial memory caused by aging, increased the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), increased total antioxidant capacity (T-AOC), and reduced the content of malondialdehyde (MDA). On the other hand, phlorizin increased the levels of interleukin-2 (IL-2) and acetylcholine (ACh), reduced the release of interleukin-6 (IL-6), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and decreased the activity of acetylcholinesterase (AChE) in the brain, improved the expression of antioxidant genes related to the nuclear factor E2-related factor 2 (Nrf2) pathway, and reduced the occurrence of morphological lesions in the hippocampus and liver. In addition, phlorizin improved cell viability and reduced the cytotoxicity of d-galactose-induced oxidative stress in PC12 cells. Meanwhile, the protective effect of phlorizin was abolished in Nrf2 gene knockdown PC12 cells. Furthermore, molecular docking showed that phlorizin could bind Keap1 protein, which can interact with Nrf2 protein. Therefore, these results suggest that phlorizin may delay senescence and enhance antioxidant capacity through the Nrf2 pathway.

Ursolic acid alleviates lipid accumulation by activating the AMPK signaling pathway in vivo and in vitro.

The mechanism underlying the effect of ursolic acid (UA) on lipid metabolism remains unclear. This study aimed to explore the mechanisms of UA in reducing lipid accumulation in free fatty acids-cultured HepG2 cells and in high-fat-diet-fed C57BL/6J mice. In vivo, UA effectively alleviated liver steatosis and decreased the size of adipocytes in the epididymis. It also significantly decreased the total cholesterol (TC) and triglyceride (TG) contents in the liver and plasma in C57BL/6 mice. In vitro, UA (20 µM) significantly reduced lipid accumulation; the intracellular TC contents decreased from 0.078 ± 0.0047 to 0.049 ± 0.0064 µmol/mg protein, and TG contents from 0.133 ± 0.005 to 0.066 ± 0.0047 µmol/mg protein, in HepG2 cells. Furthermore, UA reduced the mRNA expression related to fat synthesis, enhanced the mRNA expression related to adipose decomposition, and dramatically upregulated the protein expression of P-AMPK in vivo and in vitro. Of note, these protective effects of UA on a high-fat environment were blocked by the AMPK inhibitor (compound C) in vitro. In addition, the molecular docking results suggested that UA could be docked to the AMPK protein as an AMPK activator. These results indicated that UA lowered the lipid content probably via activating the AMPK signaling pathway, thereby inhibiting lipid synthesis and promoting fat decomposition. PRACTICAL APPLICATION: Ursolic acid (UA) widely exists in vegetables and fruits. This study highlighted a lipid-lowing mechanism of UA in HepG2 cells and C57BL/6J mice. The data indicated that UA might be used in lipid-lowering functional foods.

Dietary Supplementation of Black Rice Anthocyanin Extract Regulates Cholesterol Metabolism and Improves Gut Microbiota Dysbiosis in C57BL/6J Mice Fed a High-Fat and Cholesterol Diet.

SCOPE: This study explores the beneficial effects of dietary supplementation of black rice anthocyanin extract (BRAE) on cholesterol metabolism and gut dysbiosis. METHODS AND RESULTS: C57BL/6J mice are grouped into the normal chow diet group (NCD), the high-fat and the cholesterol diet group (HCD), and three treatment groups feeding HCD supplemented with various dosage of BRAE for 12 weeks. Results reveal that BRAE alleviates the increased body weight, serum triglyceride (TG), total cholesterol (TC), non-high-density lipoprotein cholesterol levels (non-HDL-C), and increased fecal sterols excretion and caecal short-chain fatty acids (SCFAs) concentration in HCD-induced hypercholesterolemic mice. Moreover, BRAE decreases hepatic TC content through the fundamental regulation of body energy balance gene, adenosine 5'-monophosphate activated protein kinase α (AMPKα). Meanwhile, BRAE improves the genes expression involved in cholesterol uptake and efflux, and preserves CYP7A1, ATP-binding cassette subfamily G member 5/8 mRNA expression, and the relative abundance of gut microbiota. Additionally, the antibiotic treatment experiment indicates that the beneficial effects of BRAE in reducing hypocholesterolemia risk largely depends on the gut microbiota homeostasis. CONCLUSION: BRAE supplement could be a beneficial treatment option for preventing HCD-induced hypocholesterolemia and related metabolic syndromes.

Temperature-mediated responses of carbon fluxes to precipitation variabilities in an alpine meadow ecosystem on the Tibetan Plateau.

Effects of climate warming and changing precipitation on ecosystem carbon fluxes have been intensively studied. However, how they co-regulate carbon fluxes is still elusive for some understudied ecosystems. To fill the gap, we examined net ecosystem productivity (NEP), gross ecosystem productivity (GEP,) and ecosystem respiration (ER) responses to multilevel of temperature increments (control, warming 1, warming 2, warming 3, warming 4) in three contrasting hydrological growing seasons in a typical semiarid alpine meadow. We found that carbon fluxes responded to precipitation variations more strongly in low-level warming treatments than in high-level ones. The distinct responses were attributable to different soil water conditions and community composition under low-level and high-level warming during the three growing seasons. In addition, carbon fluxes were much more sensitive to decreased than to increased precipitation in low-level warming treatments, but not in high-level ones. At a regional scale, this negative asymmetry was further corroborated. This study reveals that future precipitation changes, particularly decreased precipitation would induce significant change in carbon fluxes, and the effect magnitude is regulated by climate warming size.

Interaction mechanism of carnosic acid against glycosidase (α-amylase and α-glucosidase).

Inhibition the activity of glycosidase is an effective method for the treatment and prevention of diabetes. In this study, enzymatic kinetics, fluorescence spectrum experiment, starch granule digestion, molecular docking studies and animal's studies were used to investigate the interaction mechanism of carnosic acid against two glycosidase (α-amylase and α-glucosidase). Enzymatic kinetics showed that carnosic acid inhibited α-amylase activity in a competitive manner and α-glucosidase activity in a non-competitive manner. The half inhibitory concentrations (IC50) of carnosic acid to α-amylase and α- glucosidase were (1.12 ±â€¯0.31) and (0.08 ±â€¯0.17), respectively. The fluorescence quenching experiments showed that the intrinsic fluorescence of α-amylase or α-glucosidase was quenched by forming a complex with carnosic acid, and there was only one binding site between carnosic acid and glycosidase. The starch granules were no longer hydrolyzed by α-amylase after the addition of carnosic acid, which indicated that carnosic acid inhibited the activity of α-amylase. Molecular docking study showed that carnosic acid binds to the amino acid residues of glycosidase through hydrogen bond and van der Waals force, which leads to the change of the molecular conformation of glycosidase and thus reduces the activity of glycosidase. The experiment on mice showed that carnosic acid could effectively reduce postprandial blood glucose in mice.

Nonlinear response of ecosystem respiration to multiple levels of temperature increases.

Global warming exerts profound impacts on terrestrial carbon cycles and feedback to climates. Ecosystem respiration (ER) is one of the main components of biosphere CO2 fluxes. However, knowledge regarding how ER responds to warming is still lacking. In this study, a manipulative experiment with five simulated temperature increases (+0℃ [Control], +2.1℃ [warming 1, W1], +2.7℃ [warming 2, W2], +3.2℃ [warming 3, W3], +3.9℃ [warming 4, W4]) was conducted to investigate ER responses to warming in an alpine meadow on the Tibetan Plateau. The results showed that ER was suppressed by warming both in dry and wet years. The responses of ER to warming all followed a nonlinear pattern. The nonlinear processes can be divided into three stages, the quick-response stage (W1), stable stage (W1-W3), and transition stage (W4). Compared with the nonlinear model, the linear model maximally overestimated the response ratios of ER to warming 2.2% and 3.2% in 2015 and 2016, respectively, and maximally underestimated the ratio 7.0% and 2.7%. The annual differences in ER responding to warming were mainly attributed to the distinct seasonal distribution of precipitation. Specially, we found that the abrupt shift response of ER to warming under W4 treatment in 2015, which might be regulated by the excitatory effect of precipitation after long-term drought in the mid-growing season. This study highlights the importance of the nonlinearity of warming effects on ER, which should be taken into the global-C-cycling models for better predicting future carbon-climate feedbacks.

Effects of short-term grazing exclusion on plant phenology and reproductive succession in a Tibetan alpine meadow.

Grazing exclusion (GE) has been widely considered as an effective avenue for restoring degraded grasslands throughout the world. GE, via modifying abiotic and biotic environments, inevitably affects phenological development. A five-year manipulative experiment was conducted in a Tibetan alpine meadow to examine the effects of GE on phenological processes and reproductive success. The study indicated that GE strongly affected phenological development of alpine plant species. Specifically, the low-growing, shallow-rooted species (LSS), such as Kobresia pygmaea, are more sensitive to GE-caused changes on upper-soil moisture and light. GE advanced each phonological process of K. pygmaea, except in the case of the treatment of fencing for 5 years (F5), which postponed the reproductive stage and lowered the reproductive success of K. pygmaea. Increased soil moisture triggered by GE, especially in the upper soil, may stimulate growth of LSS. However, the thick litter layer under the F5 treatment can influence the photoperiod of LSS, resulting in suppression of its reproductive development. These findings indicate that plant traits associated with resource acquisition, such as rooting depth and plant height, mediate plant phenology and reproductive responses to grazing exclusion treatments.