Lactobacillus plantarum AR113 attenuates liver injury in D-galactose-induced aging mice via the inhibition of oxidative stress and endoplasmic reticulum stress

Probiotics could effectively eliminate excess reactive oxygen species(ROS)generated during aging or lipid metabolism disorders,but their mechanism is unclear.The major purpose of this study was to investigate the mechanism of Lactiplantibacillus plantarun AR113 alleviating oxidative stress injury in the D-galactose induced aging mice.The result showed that pretreatment with L.plantarun AR113 significantly relieving H_(2)O_(2)induced cytotoxicity in HepG2 cells by maintain cell membrane integrity and increasing antioxidant enzyme activities.In D-galactose induced aging mice,L.plantarun AR113 could significantly attenuate liver damage and inflammatory infiltration by promoting endogenous glutathione(GSH)synthesis and activating the Nrf2/Keap1 signaling pathway in mice,and increasing the expression of regulated phaseⅡdetoxification enzymes and antioxidant enzymes.Further analysis shown that gavage of L.plantarun AR113 could significantly reduce the expression of G protein-coupled receptor 78(GPR78)and C/EBP homologous protein(CHOP)proteins,and promote the restoration of endoplasmic reticulum(ER)homeostasis,thereby activating cell anti-apoptotic pathways.These results were also confirmed in H_(2)O_(2)-treated HepG2 experiments.It indicated that L.plantarun AR113 could inhibit D-galactose-induced liver injury through dual inhibition of ER stress and oxidative stress.L.plantarun AR113 have good application potential in anti-aging and alleviating metabolic disorders.

Inhibiting Effects of Achyranthes Bidentata Polysaccharide and Lycium Barbarum Polysaccharide on Nonenzyme Glycation in D-galactose Induced Mouse Aging Model

Objective To investigate the inhibiting effects and mechanism of achyranthes bidentata polysaccharide (ABP) and lycium barbarum polysaccharide (LBP) on nonenzyme glycation in D-galactose induced mouse aging model. Methods Serum AGE levels were determined by AGE-ELISA, MTT method was used to determine lymphocyte proliferation, IL-2 activity was determined by a bioassay method. Spontaneous motor activity was used to detect mouse's neuromuscular movement, latency of step-through method was used to examine learning and memory abilities of mouse, colormetric assay was used to determine hydroxyproline concentration in mouse skin, pyrogallol autoxidation method was used to determine superoxide dismutase (SOD) activity of erythrocytes. Results Decreased levels of serum AGE, hydroxyproline concentration in mouse skin and spontaneous motor activity in D-galactose mouse aging model were detected after treated with ABP or LBP, while lymphocyte proliferation and IL-2 activity, learning and memory abilities, SOD activity of erythrocytes, were enhanced. Conclusions ABP and LBP could inhibit nonenzyme glycation in D-galactose induced mouse aging model in vivo and ABP has a better inhibiting effect than LBP.

Protective Role of Salidroside against Aging in A Mouse Model Induced by D-galactose

Objective To investigate the protective effects of putative AGEs （advanced glycation endproducts） inhibitor salidroside against aging in an accelerated mouse aging model induced by D-galactose. Methods A group of 5-month-old C57BL/6J mice were treated daily with D-galactose, D-galactose combined with salidroside, salidroside alone, and control buffer for 8 weeks. At the end of the treatment, serum AGEs levels, neurological activities, expression of glial fibrillary acidic protein （GFAP） and neurotrophin-3 （NT-3） in the cerebral cortex, as well as lymphocyte proliferation and IL-2 production were determined. Results D-galactose induced mouse aging model was developed as described before. As expected, salidroside blocked D-galactose induced increase of serum AGEs levels. It also reversed D-galactose induced aging effects in neural and immune system, as evidenced by improving motor activity, increasing memory latency time, and enhancing lymphocyte mitogenesis and interleukin-2 （IL-2） production. Furthermore, elevated expression of GFAP and NT-3 in the aged model mice was also reduced upon salidroside treatment. Conclusion Salidroside inhibits AGEs formation in vivo, which at least partially contributes to its anti-aging effect in D-galactose induced aging model.

PROTECTIVE EFFECT OF TETRAMETHYLPYRAZINE ON LEARNING AND MEMORY FUNCTION IN D-GALACTOSE-LESIONED MICE

To explore the protective effect of tetramethylpyrazine (TMP) on the learning and memory function in D-galactose (D-gal)-lesioned mice. Methods C57BL/6 mice were injected (s.c.) 2% D-gal for 40 days (100 mg·kg-1·d-1). Normal saline, TMP, and Huper-zine A were respectively given by intragastric administration in different groups from the third week. Learning and memory ability was tested with Morris water maze for 5 days at the sixth week. After completion of behavioral test, the mice were sacrificed by decapitation. The brain was rapidly removed, and the cortex and hippocampus were separated. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the cortex were determined. At the same time, the activity of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), the binding sites (Bmax) and the affinity (KD) of M-cholinergic receptor in the cortex, and Bmax and KD of N-methyl-D-aspartate (NMDA) receptor in the hippocampus were determined. Results In this model group, (1) The deficit of learning and memory ability, (2) elevated MDA content and lowered SOD activity, (3) decreased AChE activity and M-cholinergic receptor binding sites in the cortex, and (4) lowered NMDA receptor binding sites were observed in the hippocampus, as compared with the normal control. TMP could markedly (1) attenuate cognitive dysfunction, (2) lower MDA content and elevate SOD activity, (3) increase the activity of ChAT and AChE, and M-cholinergic receptor binding sites in the cortex in the mice treated with D-gal. NMDA receptor binding sites were also increased in the hippocampus in the treated mice. Conclusion TMP can significantly strengthen antioxidative function, improve central cholinergic system function, protect NMDA receptor activity, and thus enhance the learning and memory ability in D-gal-lesioned mice.

Ginkgo biloba leaf extract improves the cognitive abilities of rats with D-galactose induced dementia

Standardized Ginkgo biloba leaf extract has been used in clinical trials for its beneficial effects on brain func- tions, particularly in dementia. Substantial experimental evidences indicated that Ginkgo biloba leaf extract （EGB） protected neuronal cells from a variety of insults. We investigated the effect of EGB on cognitive ability and protein kinase B （PKB） activity in hippocampal neuronal cells of dementia model rats. Rats received an intra- peritoneal injection of D-galactose to induce dementia. Forty-eight Spraque-Dawley rats were randomly divided into six groups, including the control group, D-galactose group （Gal）, low-dose EGB group （EGB-L）, mid-dose EGB group （EGB-M）, high-dose EGB group （EGB-H） and treatment group. The EGB-L, EGB-M and EGB-H groups were administered with EGB and D-galactose simultaneously. Y-maze, cresyl violet staining, TUNEL assays and immunohistochemistry staining were performed to detect learning and memory abilities, morpho- logical changes in the hippocampus, neuronal apoptosis and the expressing level of phospho-PKB, respectively. Rats in the Gal group showed decreased abilities of learning and memory, and hippocampal pyramidal cell layer was damaged, while EGB administration improved learning and memory abilities. The Gal group exhibited many stained, condensed nuclei and micronuclei, either isolated or within the cytoplasm of cells （39.5 ± 1.4）. Apoptotic cells decreased in the groups of EGB-L （35.9±0.9）, EGB-M （16.8± 1.0） and EGB-H （10.1±0.8）, and there were statistical significances compared with the Gal group. Immunoreactivity of phospho-PKB was localized diffusely throughout the cytosol of cells in all groups, while the immunoreactivity of the Gal group was weak. EGB signifi- cantly attenuated learning and memory impairment in a dose-dependent manner, while it could decrease the nmber of TUNEL-positive cells, and increase the activity of PKB. Our results demonstrated that EGB attenuated memory impairment and cell apoptosis in galactose-induced dementia model rats by activating PKB.

Moderate exercise prevents neurodegeneration in D-galactose-induced aging mice

D-galactose has been widely used in aging research because of its efficacy in inducing senescence and accelerating aging in animal models. The present study investigated the benefits of exercise for preventing neurodegeneration, such as synaptic plasticity, spatial learning and memory abilities, in mouse models of aging. D-galactose-induced aging mice were administered daily subcutaneous injections of D-galactose at the base of the neck for 10 consecutive weeks. Then, the mice were subjected to exercise training by running on a treadmill for 6 days a week. Shortened escape latency in a Morris water maze test indicated that exercise improved learning and memory in aging mice. The ameliorative changes were likely induced by an upregulation of Bcl-2 and brain-derived neurotrophic factor, the repression of apoptosis factors such as Fas and Bax, and an increase in the activity of glucose transporters-1 and 4. The data suggest moderate exercise may retard or inhibit neurodegeneration in D-galactose-induced aging mice.

Antioxidant effect of Lactobacillus fermentum HFY02-fermented soy milk on D-galactose-induced aging mouse model

This study aimed to investigate the antioxidant effect of soybean milk fermented by a new type of Lactobacillus fermentum(LF-HFY02)by using D-galactose induced aging mice model.Firstly,the optimal fermentation conditions was screened out by detecting the effects of different fermentation temperature and time on the active components and antioxidant activity of soybean milk in viro.And then unfermented soybean milk and the soybean milk fermented by different Lactobacillus was given by gavage to D-galactose-induced aging mouse.The activities of GSH,GSH-Px,SOD,CAT and T-AOC in serum,brain and liver of soybean milk fermented by LF-HFY02 were significantly increased,while the content of MDA and the level of AGEs in hippocampal were significantly decreased compared with D-galactose induced group.Further more,the mRNA expression of GSH and SOD in mouse liver were obviously up-regulated by soybean milk fermented by LF-HFY02.The skin tissue structure of mice in the LF-HFY02 fermented soybean milk group was more complete,the collagen fibers were increased and arranged orderly and liver inflammation has improved compared with the model group.And Western blot analysis showed that LF-HFY02 effectively upregulated EGFR,SOD and GSH protein expression in mouse liver.These findings suggest that LF-HFY02 can effectively prevent D-galactose-induced oxidation and aging in mice,and the effect was even better than that of the Lactobacillus delbruechii subsp.bulgaricus and vitamin C.Thus,LF-HFY02 may be potentially employed as a probiotic strain.In conclusion,soybean milk fermented by LF-HFY02 can increase the content of antioxidant factors and the activity of antioxidant enzymes by regulating gene and protein expression,and finally inhibit the process of tissue cell peroxidation,and improve the oxidative damage of mouse skin and liver.The results could provide a basis for the research and development and industrial production of probiotic-related fermented soybean milk products.

Structural and Functional Changes of Immune System in Aging Mouse Induced by D-Galactose

Objective To investigate the role of D-galactose, especially in the structural and functional changes of the immune system in aging. Methods Serum levels of advanced glycation end-products （AGE） were determined by ELISA method. Ultra-structures of thymus and spleen were detected by transmission electron microscopy. MTT method was used to determine the lymphocyte proliferation. IL-2 activity was determined by bioassay. Northern blot was used to detect the IL-2 mRNA levels. Results Serum AGE levels of D-galactose- （P〈0.01） and AGE-treated （P〈0.05） mice （n=8） were increased significantly. The ultra-structures of thymus and spleen in D-galactose- and AGE-treated mice showed regressive changes similar to those in the aged control group. The lymphocyte mitogenesis and IL-2 activity of spleen were also decreased significantly （P〈0.01, n=8）. The change of IL-2 activity shown by Northern blot resulted from the change of mRNA expression. The AGE plus aminoguanidine group, however, showed no significant change in these parameters in comparison with the young control group （P〈0.01 or P〈0.05, n=8）. Conclusion D-galactose and AGE lead to a mimic regression change of aging in the immune system in vivo.

Fingolimod(FTY720) improves postoperative cognitive dysfunction in mice subjected to D-galactose-induced aging

Neurocognitive dysfunction is a common postoperative complication,especially in older adult patients.Fingolimod(FTY720)is a sphingosine-1-phosphate receptor modulator that has been found to be neuroprotective in several animal models of central nervous system disease.However,few reports have examined whether FTY720 could mitigate postoperative cognitive dysfunction.In this study,we investigated whether FTY720 could prevent postoperative neurocognitive impairment in mice subjected to D-galactose-induced aging.We induced an accelerated model of aging by administering an intraperitoneal injection of D-galactose.Subsequently,we performed a partial hepatolobectomy under sevoflurane anesthesia.FTY720(1 mg/kg)was administered intraperitoneally 3 hours before and 24 hours after anesthesia and surgery.Our results indicated that anesthesia and surgery significantly impaired spatial memory in the Y-maze test 6 hours after surgery.We also found that problem solving ability and long-term memory in the puzzle box test on postoperative days 2–4 were significantly improved by FTY720 treatment.Immunohistochemical staining and western blot assay demonstrated that FTY720 significantly inhibited microglial activation in the hippocampal CA1 region of mice 6 hours and 3 days after anesthesia,and down-regulated the expression of synaptic-related proteins postsynaptic density protein 95 and GluR2 in the hippocampus.These results indicate that FTY720 improved postoperative neurocognitive dysfunction in mice subjected to D-galactose-induced aging.This study was approved by the Experimental Animal Ethics Committee of the Third Xiangya Hospital of Central South University of China(approval No.LLSC(LA)2016-025)on September 27,2016.

Effect of Schisandra chinensis polysaccharide on intracerebral acetylcholinesterase and monoamine neurotransmitters in a D-galactose-induced aging brain mouse model

BACKGROUND： The most prominent characteristic of brain aging is decreased learning and memory ability. The functions of learning and memory are closely related to intracerebral acetylcholinesterase （ACHE） and monoamine neurotransmitter activity. Previous studies have shown that Schisandra chinensis polysaccharide has an anti-aging effect. OBJECTIVE： To explore the effects of Schisandra chinensis polysaccharide on AChE activity and monoamine neurotransmitter content, as well as learning and memory ability in a D-galactose-induced aging mouse brain model compared with the positive control drug Kangnaoling. DESIGN, TIME AND SETTING： Completely randomized, controlled experiment based on neurobiochemistry was performed at the Pharmacological Laboratory, Henan University of Traditional Chinese Medicine from September to December 2003. MATERIALS： Schisandra chinensis was purchased from Henan Provincial Medicinal Company. Schisandra chinensis polysaccharide was obtained by water extraction and alcohol precipitation. Kangnaoling pellets were provided by Liaoning Tianlong Pharmaceutical （batch No. 20030804; state drug permit No. H21023095）. A total of 50 six-week-old Kunming mice were randomly divided into five groups： blank control, model, Kangnaoling, high and low dosage Schisandra chinensis polysaccharide groups, with 10 mice per group. METHODS： Mice in the blank control group were subcutaneously injected with 0.5 mL/20 g normal saline into the nape of the neck each day, while the remaining mice were subcutaneously injected with 5% D-galactose saline solution （0.5 mL/20 g） in the nape for 40 days to induce a brain aging model. On day 11, mice in the high and low dosage Schisandra chinensis polysaccharide groups were intragastrically infused with 20 mg/mL and 10 mg/mL Schisandra chinensis polysaccharide solution （0.2 mL/10 g）, respectively. Mice from the Kangnaoling group were intragastrically infused with 35 mg/mL Kangnaoling suspension （0.2 mL/10 g）, and the mice in the model group were intragastrically infused with the same volume of normal saline （0.2 mL/10 g） once per day for 30 consecutive days. MAIN OUTCOME MEASURES： Two hours after the final administration, pathohistological changes in the cerebral cortex and hippocampus were observed using hematoxylin ＆ eosin staining. AChE activity was detected using chromatometry. Monoamine neurotransmitter content was measured using fluorimetry. Learning and memory was measured using the step down test and darkness avoidance test. RESULTS： Both Schisandra chinensis polysaccharide and Kangnaoling improved pathological injury to the cerebral cortex and hippocampus in a mouse model of brain aging. Compared with the blank control group, AChE activity and content of norepinephrine （NA）, dopamine （DA）, and 5-hydroxytryptamine （5-HT） were significantly decreased in the model group （P 〈 0.01 ）. In contrast, AChE activity and NA, DA, and 5-HT levels significantly increased in the Kangnaoling and high dosage Schisandra chinensis polysaccharide groups （P 〈 0.01）, while NA levels significantly increased in the low dosage Schisandra chinensis polysaccharide group （P 〈 0.01）. Drug treatment improved learning and memory abilities （P 〈 0.01 or P 〈 0.05）. CONCLUSION： Schisandra chinensis polysaccharide significantly increased levels of central neurotransmitters and improved learning and memory in a mouse model of brain aging. The effects of Schisandra chinensis polysaccharide were equal to that of Kangnaoling pellets.

NADPH Oxidase-dependent Oxidative Stress and Mitochondrial Damage in Hippocampus of D-galactose-induced Aging Rats

Mitochondrial DNA(mtDNA) common deletion(CD) plays a significant role in aging and age-related diseases.In this study,we used D-galactose(D-gal) to generate an animal model of aging and the involvement and causative mechanisms of mitochondrial damage in such a model were investigated.Twenty 5-week-old male Sprague-Dawley rats were randomly divided into two groups:D-gal group(n=10) and control group(n=10).The quantity of the mtDNA CD in the hippocampus was determined using a TaqMan real-time PCR assay.Transmission electron microscopy was used to observe the mitochondrial ultrastructure in the hippocampus.Western blot was used to detect the protein levels of NADPH oxidase(NOX) and uncoupling protein 2(UCP2).We found that the level of mtDNA CD was significantly higher in the hippocampus of D-gal-induced aging rats than in control rats.In comparison with the control group,the mitochondrial ultrastructure in the hippocampus of D-gal-treated rats was damaged,and the protein levels of NOX and UCP2 were significantly increased in the hippocampus of D-gal-induced aging rats.This study demonstrated that the levels of mtDNA CD and NOX protein expression were significantly increased in the hippocampus of D-gal-induced aging rats.These findings indicate that NOX-dependent reactive oxygen species generation may contribute to D-gal-induced mitochondrial damage.

Protective effects of (-)-epigallocatechin-3-gallate on D-galactose-induced neuronal apoptosis in mice

BACKGROUND： The neuroprotective effects of （-）-epigallocatechin-3-gallate （EGCG）, the main polyphenolic constituent of green tea, have been widely reported. However, the action mechanisms, in particular in D-galactose-induced aging mice, remain poorly understood. OBJECTIVE： The present study investigated the protective effects of EGCG on D-galactose-induced hippocampus neuronal apoptosis in aging mice, as well as the relationship with expression of p751CD, JNK2, and p53 proteins. DESIGN, TIME AND SETTING： A randomized, controlled, molecular biological, animal experiment was performed at the Laboratory of Pharmacology, Pharmaceutical College of China Medical University, China, from September 2006 to July 2008. MATERIALS： D-galactose and EGCG （Sigma, USA）, as well as terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling （TUNEL） In Situ Cell Apoptosis Detection Kit （Promega, USA）, were used in this study. METHODS： A total of 64 mice were equally and randomly divided into D-galactose model, low-dose EGCG, high-dose EGCG, and control groups. Mice in the D-galactose model, low-dose EGCG, and high-dose EGCG groups were subcutaneously injected with 3% D-galactose （150 mg/kg）, daily for 6 weeks, to establish a mouse model of aging. Mice in the control group were treated with saline （5 mL/kg）. At 3 weeks following injection, mice in the low-dose EGCG and high-dose EGCG groups were orally administered EGCG at a dose of 2 mg/kg and 6 mg/kg, respectively, once a day, for 4 consecutive days. Mice in the control and D-galactose model groups received distilled water （5 mL/kg）. MAIN OUTCOME MEASURES： Memory function was evaluated using a step-through passive avoidance test. Neuronal apoptosis in the mouse hippocampus was detected using TUNEL staining. Expression levels of the intracellular domain of the p75 neurotrophin receptor （p75NTR）-p751CD, JNK2, and p53 proteins in the hippocampus were determined using Western blot analysis. RESULTS： The aging mouse model was induced by subcutaneous injection of D-galactose, which resulted in obvious memory impairment, increased apoptotic index, and increased protein expression levels of p751CD, JNK2, and p53 in the hippocampus, compared with control mice （P 〈 0.01）. Oral EGCG administration （2 or 6 mg/kg） for 4 weeks significantly improved levels of memory deficit in the aging mice and reduced apoptotic indices and protein expression levels of p751CD, JNK2, and p53 in the mouse hippocampus （P 〈 0.01）. CONCLUSION： Results from this study demonstrated increased protein expression levels of p751CD, JNK2, and p53, as well as increased hippocampal neuronal apoptosis in a D-galactose-induced mouse model of aging. EGCG provided protective effects against D-galactose-induced neuronal apoptosis in the hippocampus by reducing protein expression levels of p751CD, JNK2, and p53 proteins in the hippocampus of aging mice.

The Antioxidant Effects of L-carnitine on D-galactose Induced Aging Mice

[Objective] To study the antioxidant effects of LC on D-galactose induced aging mice. [Method] 50 mice were randomly divided into blank control group, D-galactose model group, LC treatment groups at three levels of 0.25, 0.5 and 1.0 g/（kg d）, a total of 5 groups. Using the subacute D-galactose induced aging models, the mice were killed after 6 weeks of experiment and the activity of Superoxide dismutase （SOD）, Glutathione peroxidase （GSH-Px） and Malondialdehyde （MDA） content in plasma, brain and liver tissue were determined. [ Result.] The activity of SOD and GSH-Px in plasma, brain and liver tissue were significantly increased after treated with LC, while the content of MDA were decreased with a dose-dependent manner. [ Conclusion] LC had anti-aging effects in mice, and the mechanism may be correlated with the antioxidation.

Study on Learning and Memory Ability of Mice Induced by Different Doses of D-galactose

Aim:To explore the influence of different doses of D-galactose on learning memory capacity in mice.Methods:The 2-month-old KM mice were randomly divided into D-galactose-treated groups (75,100,150,300 mg·kg^-1) and normal control group,with 12 mice in each group.The corresponding dose of D-galactose was administered subcutaneously to the back of the neck,and normal control group was injected subcutaneously with saline for 8 weeks.Learning memory capacity of mice was detected through Morris water maze,step-down passive avoidance test and new object recognition test.Meanwhile,the enzymatic indicators in the brain were determined to detect the damage degree in mice with different doses of D-galactose.Results:Compared with the normal control group,the spatial and non-spatial cognitive functions and brain enzyme levels of D-galactose in each dose group was damaged to varying degrees,especially the dose of 100 mg·kg-1 group.In the water maze experiment,D-galactose groups significantly prolonged the time of crossing the platform for the first time and decreased the number of crossing platforms,especially at the dose of 100 mg·kg-1 (P < 0.05);in the new object recognition experiment,discrimination index was significantly decreased in each D-galactose group (P < 0.01),and the dose of 100 mg·kg-1 was most obvious;in addition,D-galactose could significantly increase the level of acetylcholinesterase (AChE)(P < 0.01),decrease the levels of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px),and total antioxidant capacity (T-AOC)(P < 0.05 or P < 0.01) in the mice brain.Conclusion:Subcutaneous injection of low-medium-high dose of D-galactose for 8 weeks in the back of the neck can cause changes in brain parameters in mice,but the effects of different doses of D-galactose on some indicators are significantly different.D-galactose model is effective and stable at the dose of 100 mg·kg-1.

Effect of Ningxin-Yishen Formula on D-galactose-induced Premature Ovarian Insufficiency Mice by Inhibiting p53

Background:The prevalence of premature ovarian insufficiency(POI)is gradually increasing,safer and more effective treatments are urgently needed.Objective:The aim of this study was to evaluate the effects of Ningxin-Yishen formula(NXYSF)on D-galactose-induced POI mice as well as to shed a light on its potential mechanisms.Methods:Six to eight weeks old female C57BL/6 mice were used in this study and randomly divided into six groups:control group;model group;estradiol valerate(EV)treatment group and NXYSF treatment group with graded doses(9.5,19,and 38 g·kg^(−1)/d).Both EV and NXYSF treatments were initiated at the 15th day of modeling and lasted for 28 days.Afterwards,the ovarian function was evaluated in each group by analyzing the proportion of primordial follicles as well as the serum sex hormone levels.Furthermore,network pharmacology approach was performed to elucidate the potential targets of NXYSF,which was verified through western blotting experiments finally.Results:NXYSF could significantly reverse the inefficiency of weight gain caused by POI,and promote the devel-opment of primordial follicles.In addition,it could restore the abnormal serum anti-Müllerian hormone(AMH),estradiol(E 2),luteinizing hormone(LH)and follicle-stimulating hormone(FSH).Moreover,some crucial key gene targets including TP53 were as propose to be relate with the effect of NXYSF through network pharmacology anal-ysis.Last but importantly,western blotting experiments confirmed that NXYSF could inhibit the expression of p53 protein in mouse ovaries.

Combined administration of D-galactose and aluminium induces Alzheimer-like lesions in brain

Objective It has been reported that D-galactose （D-gal） can model subacute aging, and aluminum （AI） acts as a neurotoxin, but combined effects of them have not been reported. The present work aimed to reveal the effect of combined administration of D-gal and A1 in mice and compare the effect of D-gal treatment with that of A1 treatment. Meth- ods A1 was intragastricaHy administered and D-gal was subcutaneously injected into Kunming mice for 10 consecutive weeks. Learning and memory, eholinergic systems, as well as protein levels of amyloid β （Aβ） and hyperphosphorylated tau were determined using Morri water maze test, biochemical assays and immunohistochemical staining, respectively. Results The mice with combined treatment had obvious learning and memory deficits, and showed decreases in brain ace- tylcholine （ACh） level and in activities of choline acetyltransferase （CHAT） and acetyleholinesterase （ACHE）. Formation of senile plaque （SP）-like and neurofibrillary tangle （NFT）-like structures was also observed. The behavioral and pathologi- cal changes persisted for at least 6 weeks after withdrawal of D-gal and A1. Conclusion Combined use of D-gal and A1 is an effective way to establish the non-transgenic Alzheimer＇s disease （AD） animal model, and is useful for studies of AD pathogenesis and therapeutic evaluation.

Low-temperature Heat Capacity and Standard Thermodynamic Functions of D-Galactose and Galactitol

The heat capacities of D-galactose and galactitol were measured on a quantum design physical property measurement system（PPMS） over a temperature range of 1.9-300 K, and the experimental data were fitted to a function of T using a series of theoretical and empirical models in appropriate temperature ranges. The fit results were used to calculate thermodynamic function values, C^θp,m, ^T0S0^θm , and △^T0H^θm from 0 K to 300 K. The standard molar heat capacity, entropy and enthalpy values of D-galactose and galactitol at 298.15 K and 0.1 MPa were determined to be C^θp,m=（227.96±2.28） and（239.50±2.40） J·K^-1·mol^-1, S0^θm = （211.22±2.11） and （230.82±2.30） J·K^-1·mol^-1 and μm = （33.95±0.34） and （36.57±0.37） kJ/mol, respectively.

有氧训练对D-galactose模型大鼠生长状况影响的实验研究

本研究选用离乳6周龄雄性SD大鼠40只，研究运动对衰老模型大鼠体重、体长、饮食量扣相关脏器指数的影响。结果发现有氧训练可以改善机体的机能水平，具有延缓机体衰老作用。特别的隔天训练要比每天训练更能改善机体的机能水平，有效延缓机体衰老。

Multi-protective effects of wheat embryo globulin on D-gal-induced aging mice

Wheat embryo globulin(WEG)has been proven to possess multiple biological activities,including antioxidative properties,immunomodulatory,and so on.Aged mouse model were established by subcutaneous injection of D-galactose(D-gal),and the effects of WEG on learning,memory,and antioxidant capacity in aging mice were explored through behavioural tests and antioxidant enzyme activities determination.Compared with the Model group,WEG improved the percentage of the platform quadrant,increased the number of crossing platforms,and enhanced the identification indexs.WEG also increased total antioxidant capacity(T-AOC),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)activities in the liver and brains of aging mice,and reduced malondialdehyde(MDA)content.Pathological observations indicated that WEG protected against damage to brain in D-gal-induced aging mice.These results effectively revealed that WEG not only improved the abilities of learning and memory,and the cognitive impairment,but also delayed the aging process of the D-gal-induced mice.

Establishment of oxygen glucose deprivation reperfusion model of senescent SH-SY5Y cells

Obejective:To explore the establishment of an oxygen glucose deprivation/reperfusion model of senescent SH-SY5Y cells.Methods:SH-SY5Y cells were randomly divided into control(D-galactose 0 mmol/L group),D-galactose(25 mmol/L,50 mmol/L,100 mmol/L,200 mmol/L,400 mmol/L)groups,and treated with corresponding concentrations of D-galactose for 48 h.The changes of cell morphology,β-galactosidase,the cell morphology,β-galactosidase activity by microscopic observation,cell proliferation rate by EdU kit and cell survival rate by CCK-8 assay were used to determine the decaying concentration of D-galactose and to establish the senescence model.The senescent SH-SY5Y cells were randomly divided into control group(oxygen glucose deprivation without treatment group),oxygen glucose deprivation treatment(0.5 h,1 h,1.5 h,2 h)group,followed by re-glucose reoxygenation for 24 h,and CCK-8 assay for the survival rate of senescent SH-SY5Y cells.Results:There were no significant changes in cell morphology and β-gal activity in the 25 mmol/L and 50 mmol/L groups compared with the control group(P>0.05),cytosolic hypertrophy was seen in the cells of the 100 mmol/L group,chromatin fixation in the cells of the 200 mmol/L group,and massive vacuolization in the cells of the 400 mmol/L group;the positive rate ofβ-galactosidase staining in the cells of the(100-400 mmol/L)group was significantly higher compared with the control group(P<0.05),with little difference between the 100 mmol/L and 200 mmol/L groups(P>0.05);the cell proliferation ability of the(100-400 mmol/L)group was significantly decreased in a concentration-dependent manner(P<0.05);the cell survival rate was decreased in a concentration-dependent manner(P<0.05),with IC_(50) between 100 mmol/L and 200 mmol/L.The survival of senescent SH-SY5Y cells showed a time-dependent decrease in oxygen-glucose deprivation(P<0.05),with an IC_(50) close to 1 h.Conclusion:D-gal concentration of 100 mmoL/L and 48 h of cell action could establish a survival rate of about 50%of senescent SH-SY5Y cells,and oxygen glucose deprivation of senescent SH-SY5Y cells for 1 h and reperfusion for 24 h could establish an oxygen glucose deprivation/reperfusion model of senescent SH-SY5Y cells with a survival rate close to 50%.