D-galactose Group Research Articles

THE EFFECTS OF OXIDATIVE STRESS ON THE EYES OF SPRAGUE–DAWLEY RATS

An imbalance between oxidants and antioxidants in favor of the oxidants, potentially leading to damage, is termed as ‘oxidative stress’. Some studies indicate that reactive oxygen species (ROS) may increase in resistance of the aqueous humor drainage or induce the rearrangement of human trabecular meshwork and result in glaucoma finally. An eight-week d-galactose injection was used to mimic high oxidative stress rats. Behavior tests, fundus images, anterior chamber depth, anterior chamber diameter (ACDia) and pupil diameter (PD) were investigated. Intraocular pressure (IOP) measurement and serum were taken for antioxidant capability of superoxide dismutase (SOD) and the amount of nitric oxide (NO). After perfusion, Müller’s cells and astrocyte, microglial and inducible Nitric Oxide Synthase (iNOS) were labeled. The activities of SOD were decreased after induction. The spatial memory ability and muscular endurance were impaired through Morris water maze (MWM) and weight-loaded forced swimming test. These results demonstrated that oxidative stress was exacerbated by d-galactose. The IOP was gradually increased in the third week after d-galactose induction. Immunohistochemical stains showed that the density of Müller’s cells, astrocytes and microglia increased in d-galactose group. Microglia mostly became active. These data indicate that oxidative stress may result in the impairment of behavior, increase of Müller’s cells, astrocyte and microglia which is induced by d-galactose. The correlation between oxidative stress and elevation of IOP should be further studied. These may involve in the pathologies of aging or other neurodegenerative disease.

Effects of caloric restriction on DNA damage response-mediated senescence in a rat model of D-galactose-induced brain aging

Background Aging is a multifaceted irreversible impairment of normal function. The exact pathogenesis of aging is still unclear. However, DNA damage, cell senescence, and oxidative stress emerged as the major contributing factors in aging development. Aims The goal of this study was to assess the defensive action of caloric restriction (CR) against D-galactose-induced brain aging in rats, pointing to its ability to modulate the levels of phosphorylated histone H2AX (DNA damage response marker) and p16 (senescence marker) and oxidative stress markers. Materials and methods This study was conducted on 48 male albino rats that were allocated into four groups: group І (control group), group II (dietary-restricted group), group IIІ (D-galactose group), and group IV (dietary-restricted and D-galactose group). All rats were subjected to measurement of brain tissue levels of p16 and phosphorylated histone H2AX by enzyme-linked immunosorbent assay technique, while reduced glutathione (GSH) and malondialdehyde (MDA) levels were measured colorimetrically. Additionally, Morris water-maze test was performed to all groups, as a neurobehavioral test. Results The levels of brain tissue p16, phosphorylated histone H2AX, and MDA were increased significantly in D-galactose-injected rats relative to control rats, while the levels of reduced GSH were significantly decreased in these rats. On the other hand, CR significantly decreased the brain tissue levels of p16, phosphorylated histone H2AX, and restored oxidative balance evidenced by alleviation of the levels of MDA and increasing reduced GSH levels. Conclusion These findings hold a great promise for CR as a possible neuroprotective intervention to delay age-related deterioration of brain function.

Ceftriaxone improves hepatorenal damages in mice subjected to D-galactose-induced aging

Ceftriaxone (CTX) is a third-generation cephalosporin antibiotic that has broad-spectrum antimicrobial activity. This agent also has anti-inflammatory and antioxidant characteristics. In the current study, the effects of CTX against hepatorenal damages in a D-galactose (DGL) induced aging model were investigated. We used twenty-eight male mice which equally and randomly were separated into four groups as follows: Control, DGL group (treated with 500 mg/kg/day DGL orally for six weeks), DGL + CTX group (treated with 500 mg/kg/day DGL orally plus 200 mg/kg/day CTX intraperitoneally for six weeks), and CTX group (treated with 200 mg/kg/day CTX intraperitoneally for six weeks). The liver and kidney function indices such as serum creatinine, blood urine nitrogen, alanine aminotransferase, and aspartate aminotransferase were measured. Also, levels of malondialdehyde, catalase, and glutathione peroxidase in hepatic and renal tissues were evaluated. Moreover, the expression profiles of interleukin 1 beta and tumor necrosis factor alpha were assessed. The liver and kidney tissues were assessed for histopathological lesions. The results showed that aging induced by DGL leads to abnormalities in functional indices of the liver and kidneys. DGL also induced significant oxidative stress and inflammation, as well as histopathological lesions, in these organs. CTX improved functional indices, as well as the parameters of oxidative stress and inflammation, compared with the DGL-treated animals. These results were also confirmed by histological evaluations of the liver and kidneys. These data provide evidence for the therapeutic value of CTX in clinical practice for mitigating the hepatorenal damages of aging.

Improving glucose metabolism in the auditory cortex delays the aging of auditory function of guinea pig

The glucose homeostasis is essential for brain function, and energy deficiency is a key feature of brain aging. We investigated whether improving glucose metabolism in the auditory cortex can delay the aging of auditory function of guinea pigs with age-related hearing loss (ARHL) by d-galactose. Auditory function was assessed by auditory brainstem response (ABR), glucose metabolism was detected by micro PET/CT, and the proteome were identified in auditory cortex by two-dimensional electrophoresis and matrix assisted laser desorption/ionization mass spectrometry. Glucose metabolism decreased in the auditory cortex of d-galactose group, and improving glucose metabolism can delay the aging of auditory function by upregulating seven metabolism-related proteins including ATP synthase subunit beta, triosephosphate isomerase, creatine kinase U-type, pyruvate dehydrogenase E1 component subunit beta, alpha-enolase, phosphoglycerate kinase, and tubulin beta-2A chain. These results suggest that the decrease of glucose metabolism in the auditory cortex may be an important role in the aging of auditory function, and improving glucose metabolism in the auditory cortex can delay the aging of auditory function of guinea pig with ARHL induced by d-galactose.

Ceftriaxone improves senile neurocognition damages induced by D-galactose in mice.

Objective(s):Ceftriaxone (Cef), a beta-lactam antibiotic, is accompanied by antioxidant and anti-inflammatory properties. It has been shown that Cef has beneficial effects on Alzheimer’s disease. In the current investigation, the effect of Cef in a mice model of aging was investigated. Materials and Methods:Forty male mice were equally aliquoted into four groups as follows: Control (as healthy normal animals), D-galactose (DG) group (treated with 500 mg/kg/day DG for 6 weeks), DG + Cef group (treated with DG plus Cef 200 mg/kg/day for 6 weeks), and Cef group (treated with Cef 200 mg/kg/day for 6 weeks). A battery of behavioral tests was done to evaluate age-related neurocognitive changes. The activities of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD), as well as the level of malondialdehyde (MDA) in the brain, were measured by biochemical methods. Also, to determine the brain damage, histopathological alterations in the hippocampus were measured using hematoxylin and eosin (H&E) staining. Results:Our results indicate that neurobehavioral dysfunctions of DG can be prevented by co-administration of Cef. We also found that Cef increases the activity of SOD, GPx, and CAT as well as decreasing the level of MDA in the brain of aged mice. Conclusion:Based on our findings, Cef declines neurocognitive dysfunctions in the DG-induced model of aging, possibly through its antioxidative properties.

Rare Ginsenoside 20(R)-Rg3 Inhibits D-Galactose-Induced Liver and Kidney Injury by Regulating Oxidative Stress-Induced Apoptosis.

Oxidative stress is considered as a major factor in aging and exacerbates aging process through a variety of molecular mechanisms. D-galactose, a normal reducing sugar with high dose can cause the accumulation of reactive oxygen species (ROS) or stimulate free radical production indirectly by the formation of advanced glycation end products in tissues, finally resulting in oxidative stress. 20(R)-ginsenoside Rg3 (20(R)-Rg3), a major and representative component isolated from red ginseng (Panax ginseng C.A Meyer), has been shown to observably have an anti-oxidative effect. We thereby investigated the beneficial effects of 20(R)-Rg3 on D-galactose-induced oxidative stress injury and its underlying mechanisms. Our results showed that continuous injection of D-galactose with 800[Formula: see text]mg/kg/day for 8 weeks increased the levels of alanine aminotransferase (ALT) and blood urea nitrogen (BUN). However, such increases were attenuated by the treatment of 20(R)-Rg3 for 4 weeks. Meanwhile, 20(R)-Rg3 markedly inhibited D-galactose-caused oxidative stress in liver and kidney. The anti-oxidants, including catalase (CAT) and superoxide dismutase (SOD), were elevated in the mice from 20(R)-Rg3-treated group compared with that from D-galactose group. In contrast, a significant decrease in levels of cytochrome P450 E1 (CYP2E1) and the lipid peroxidation product malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) were observed in the 20(R)-Rg3-treated group. These effects were associated with a significant increase of AGEs. More importantly, 20(R)-Rg3 effectively attenuated D-galactose induced apoptosis in liver and kidney via restoring the upstream PI3K/AKT signaling pathway. Taken together, our study suggests that 20(R)-Rg3 may be a novel and promising anti-oxidative therapeutic agent to prevent aging-related injuries in liver and kidney.

Aging induced by D-galactose aggravates cardiac dysfunction via exacerbating mitochondrial dysfunction in obese insulin-resistant rats.

The prevalence of obesity and an aging population are increasing worldwide. Both obesity and aging are independently known to be associated with cardiac dysfunction. However, in obese insulin-resistant subjects, the effects of aging on metabolic status and cardiac and mitochondrial functions are not completely understood. We hypothesized that in the obese insulin-resistant condition, aging induced by D-galactose increases cardiac senescence markers and aggravates the impairment of metabolic parameters, cardiac and mitochondrial function, and increases oxidative stress, inflammation, apoptosis, and autophagy. Sixty-four male Wistar rats were fed with either normal diet (ND) or high-fat diet (HFD) for 12 weeks. Then, rats were divided into vehicle groups (0.9% NSS, subcutaneous injection (SC)) or D-galactose groups (150 mg/kg/day, SC). After 0.9%NSS or D-galactose treatment for 4 weeks and 8 weeks, metabolic and cardiac functions were determined. The heart was then removed to determine mitochondrial functions and enable biochemical studies. After 4 weeks of D-galactose injection, ND rats treated with D-galactose (NDD4), HFD rats treated with vehicle (HFV4), and HFD rats treated with D-galactose (HFD4) had reduced cardiac function, impaired cardiac mitochondrial function and autophagy, and increased oxidative stress, inflammation, and apoptosis. Interestingly, after 8 weeks, HFD rats treated with D-galactose (HFD8) had the worst impairment of cardiac and mitochondrial function, autophagy, and apoptosis in comparison to the other groups. Aging induced by D-galactose aggravated cardiac dysfunction in obese insulin-resistant rats through the worsening of cardiac mitochondrial function, autophagy, and increased apoptosis in a time-dependent manner.

Production of Galactose Oxidase Inside the Fusarium fujikuroi Species Complex and Recombinant Expression and Characterization of the Galactose Oxidase GaoA Protein from Fusarium subglutinans.

Galactose oxidase catalyzes a two-electron oxidation, mainly from the C6 hydroxyl group of D-galactose, with the concomitant reduction of water to hydrogen peroxide. This enzyme is secreted by Fusarium species and has several biotechnological applications. In this study, a screening of galactose oxidase production among species of the Fusarium fujikuroi species complex demonstrated Fusarium subglutinans to be the main producer. The truncated F. subglutinans gaoA gene coding for the mature galactose oxidase was expressed from the prokaryotic vector pTrcHis2B in the E. coli Rosetta™ (DE3) strain. The purified recombinant enzyme presented temperature and pH optima of 30°C and 7.0, respectively, KM of 132.6 ± 18.18mM, Vmax of 3.2 ± 0.18µmol of H2O2/min, kcat of 12,243s-1, and a catalytic efficiency (kcat/KM) of 9.2 × 104M-1s-1. In the presence of 50% glycerol, the enzyme showed a T50 of 59.77°C and was stable for several hours at pH 8.0 and 4°C. Besides D-(+)-galactose, the purified enzyme also acted against D-(+)-raffinose, α-D-(+)-melibiose, and methyl-α-D-galactopyranoside, and was strongly inhibited by SDS. Although the F. subglutinans gaoA gene was successfully expressed in E. coli, its endogenous transcription was not confirmed by RT-PCR.

Effects of different doses of curcumin on testicular histopathology, apoptosis, and reproductive organs weight index in mice d-galactose-induced aging model

Due to the value of aging and its effect on body organs, it is necessary to study the factors that can have protective effects on this process. Curcumin is the active ingredient of Curcuma longa Turmeric, which has a strong antioxidant effect. We evaluated the curcumin treatment on testicular histopathology, apoptosis, and reproductive organs index in a mouse model of aging. Forty-eight mice divided into control, sham, d-galactose, and curcumin 1 to 3 groups. The d-galactose group was administered intraperitoneally 300 mg/kg d-galactose for 6 weeks. The curcumin 1 to 3 groups received d-galactose plus 25, 50, and 100 mg/kg of curcumin, respectively for 14 days. Then, testicular histopathology, apoptosis, and reproductive organs index were investigated after 6 weeks. The SPSS software was used for data analysis. Tunnel-positive cells in the d-galactose group were remarkably higher than the control group (p < 0.05). Administration of curcumin in all three doses reduced the number of positive tunnel cells, in comparison with the group that received d-galactose (p < 0.05). The epididymis weight index of the d-galactose group was significantly different from the control (p = 0.006), the sham (p = 0.01), the curcumin 1 (p = 0.003), the curcumin 2 (p = 0.003), and the curcumin 3 (p = 0.002) groups. Histological examination of the d-galactose group revealed a reduction in the thickness of the germinal epithelium, edema, and complete atrophy of germ cells in some seminiferous tubules. The histology of seminiferous tubules was near-normal appearance with the little disintegration of germ cells from their normal germinal epithelium in the curcumin 1 and 3 groups. The histology of seminiferous tubules was near-normal appearance with mild edema and congestion in some seminiferous tubules in the curcumin 2 group. Curcumin supplementation (2 weeks, 100 mg/kg) caused an improvement in biochemical markers and sperm parameters as well as a reduction in apoptosis in the testicular tissue of the aging mouse model.

Effects of Thymoquinone on Sperm Parameters, Apoptosis, Testosterone Level, and Oxidative Stress in a Mouse Model of D-Galactose-Induced Aging

Background: The aging process is accompanied by low secretion of sex hormones and testicular apoptosis. The antioxidant properties of thymoquinone (TQ) may prevent the effects of aging. Therefore, in the present study, the effects of different doses of TQ were investigated on sperm parameters, testosterone level, apoptosis, and oxidative stress in a mouse model of D-galactose-induced aging. Methods: In this experimental study, 30 adult male mice were randomly divided into 5 groups. The control group did not receive any injections, while the D-galactose group received an intraperitoneal injection of 300 mg/kg of D-galactose for 42 days. The TQ1-TQ3 groups received intraperitoneal injections of 5, 2.5, and 1.25 mg/kg of TQ plus D-galactose, respectively for 14 days (from the 1st to the 14th day of the experiment). The morphometric analysis, testicular apoptosis examination, and sperm analysis were performed, and testosterone level, total antioxidant capacity, and malondialdehyde level were evaluated on day 42 of the experiment. Data were analyzed using SPSS. Results: Administration of TQ in the TQ1 group caused a significant difference in sperm parameters, compared to the D-galactose group (P&lt;0.05). The lowest amount of positive tunnel cells was related to 5 mg/kg of TQ and the highest to 2.5 mg/kg of TQ. There was no significant difference in the parameters of seminal vesicles, epididymis, prostate, and testis between the groups (P&gt;0.05). The malondialdehyde level were decreased in the TQ1-TQ3 groups, compared to the D-galactose group (P&lt;0.001). On the other hand, the total antioxidant capacity was increased significantly in the TQ1 group, compared to the D-galactose group (P&lt;0.001). Conclusion: Administration of 5 mg of TQ for 14 days improved sperm quality and biochemical parameters, while reducing apoptotic cells of the testes in a mouse model of aging.

Aceclofenac–Galactose Conjugate: Design, Synthesis, Characterization, and Pharmacological and Toxicological Evaluations

Aceclofenac is a popular analgesic, antipyretic, and nonsteroidal anti-inflammatory drug (NSAID) used for prolonged treatment (at least three months) in musculoskeletal disorders. It is characterized by several limitations such as poor water solubility and low oral bioavailability. The main side-effect of aceclofenac, as well as all NSAIDs, is the gastrotoxicity; among other adverse effects, there is the risk of bleeding since aceclofenac reversibly inhibits platelet aggregation. With the aim to reduce these drawbacks, we have designed, synthesized, and characterized, both in vitro and in vivo, an orally administrable pro-drug of aceclofenac (ACEgal). ACEgal was obtained by conjugating carboxyl group with the 6-OH group of d-galactose; its structure was confirmed by X-ray powder diffractometry. The pro-drug was shown to be stable at 37 °C in simulated gastric fluid (SGF-without pepsin, pH = 1.2) and moderately stable in phosphate buffered saline (PBS, pH = 7.4). However, it hydrolyzed in human serum with a half-life ( t1/2) of 36 min, producing aceclofenac. Furthermore, if compared to its parent drug, ACEgal was four-times more soluble in SGF. To predict human intestinal absorption, cell permeability in a Caco-2 model of aceclofenac and ACEgal was determined. Anti-inflammatory, analgesic, and ulcerogenic activities have been investigated in vivo. In addition, oxidative stress parameters (thiobarbituric acid reactive substances, TBARS, and glutathione, GSH) and platelet antiaggregatory activity both of parent drug and pro-drug were evaluated. Results clearly showed that the conjugation of aceclofenac to a galactose molecule improves physicochemical, toxicological (at gastric and blood level), and pharmacological profile of aceclofenac itself without changing intestinal permeability and antiplatelet activity (in spite the new sugar moiety).

EFFECT OF HYDROALCOHOLIC EXTRACT OF PURSLANE (PORTULACA OLERACEA L.) ON DIABETIC VARIABLES IN D-GALACTOSE INDUCED AGING MOUSE MODEL.

Purslane (Portulaca oleracea L.) has antioxidant effects. The aim of this study was to evaluate the pancreas protective effect of Purslane hydroalcoholic extract in D-galactose induced aging female mouse model. In this experimental study, 72 adult female mice (30 - 35 g) were obtained and divided into 6 groups: control, Purslane hydroalcoholic extract, D-galactose, D-galactose + Purslane hydroalcoholic extract, Aging, Aging + Purslane hydroalcoholic extract. The aging model induced by subcutaneous injection of D-galactose for 45 consecutive days, and Purslane hydroalcoholic extract was orally gavaged in the last 21 days. 24 hours after the last drug and extract administrations, serum samples and pancreas tissues were removed for biochemical and histological assessments. Glucose decreased in the Purslane, D-galactose + Purslane and Aging + Purslane groups (p<0.05). Insulin and HOMA-IR increased in D-galactose and, Aging groups (p<0.05). Furthermore, administration of hydroalcoholic extract of Purslane improved these parameters in D-galactose and Aging treated mice (p<0.05). Diameter of pancreatic islets decreased in Aging and D-galactose groups and Purslane hydroalcoholic extract administration improved this variable. The present results show that Purslane has pancreas protective effects via its hypoglycemic and insulin resistance reducing activity.

Exendin-4 protects mice from D-galactose-induced hepatic and pancreatic dysfunction.

ABSTRACTInvestigations into pharmaceutical intervention of pancreatic and hepatic dysfunction associated with metabolic disturbances have received relatively little attention. The aim of this study was to investigate the protective effects of exendin-4 in mice receiving D-galactose, a reducing sugar that triggers ROS production and inflammatory mediators affecting the pancreas and liver. Exendin-4 is an United States Food and Drug Administration (FDA) approved glucagon-like peptide that increases insulin dependent glycogen synthesis and glucose uptake. Male NMRI mice (20–25 g), 3 months of age, were randomly divided into 6 groups of 12 mice each: control, exendin-4 (1 nmol/kg), exendin-4 (10 nmol/kg), D-galactose, D-galactose + exendin-4 (1 nmol/kg) and D-galactose + exendin-4 (10 nmol/kg). D-galactose (500 mg/kg) was given daily by oral gavage for 6 weeks. During the last 10 days, exendin-4 (1 and 10 nmol/kg) was injected intraperitoneally daily. Glucose, insulin, insulin resistance, lipid profiles, and hepatic enzyme levels significantly increased in the D-galactose group (p < 0.05), along with a significant decrease in superoxide dismutase activity and pancreatic islet insulin secretion (p < 0.05). Exendin-4 decreased D-galactose-induced increases in serum glucose and insulin, insulin resistance, lipid profiles, and hepatic enzymes, and improved pancreatic islet insulin secretion and antioxidant defense status. The results show that exendin-4 can prevent complications in mice with compromised pancreatic and hepatic function. Long term administration of D-galactose in mice may be a useful model to study insulin resistance, metabolic syndrome, and aging.

Pancreatic protective and hypoglycemic effects of Vitex agnus-castus L. fruit hydroalcoholic extract in D-galactose-induced aging mouse model.

D-galactose induces pancreatic disorder along with aging mouse model. Vitex agnus-castus (VAC) has potential pancreatic protective effect. Hence, this study was designed to evaluate the hypoglycemic and pancreas protective effects of VAC hydroalcoholic extract in D-galactose-induced aging female mice. In the present experimental study, 72 adult female Naval Medical Research Institute (NMRI) mice (weighing 30–35 g) were divided into 6 groups of control, VAC hydroalcoholic extract, D-galactose, D-galactose + VAC hydroalcoholic extract, aged, aged + VAC hydroalcoholic extract. The aged model was prepared by subcutaneous injection of D-galactose for 45 days and, VAC hydroalcoholic extract was gavaged twice a day in the last 7 days. 24 h after the last drug and extract administrations, serum samples and pancreatic tissues were removed to evaluate experimental and histological determinations. Serum glucose level decreased in VAC, D-galactose and, aged-treated groups compared to the control (P < 0.05). Insulin level increased in VAC and decreased in D-galactose and aged VAC-treated mice compared to the control (P < 0.05). Homeostasis model assessment-estimated insulin resistance (HOMA-IR) increased in D-galactose, aging, and VAC hydroalcoholic extract groups (P < 0.05) and, administration of VAC hydroalcoholic extract improved HOMA-IR in D-galactose and aging treated animals. Despite the size of pancreatic islets decreased in aged and D-galactose groups, VAC administration recovered it. Present data showed that VAC hydroalcoholic extract has hypoglycemic and pancreatic protective effects in natural aged and aging model mice.

EPA or DHA enhanced oxidative stress and aging protein expression in brain of d-galactose treated mice

Background:Effects of eicosapentaenoic acid (EPA, 20:5) and docosahexaenoic acid (DHA, 22:6) upon fatty acid composition, oxidative and inflammatory factors and aging proteins in brain of d-galactose (DG) treated aging mice were examined.Methods:Each fatty acid at 7 mg/kg BW/week was supplied for 8 weeks. Brain aging was induced by DG treatment (100 mg/kg body weight) via daily subcutaneous injection for 8 weeks.Results:DG, EPA and DHA treatments changed brain fatty acid composition. DG down-regulated brain Bcl-2 expression and up-regulated Bax expression. Compared with DG groups, EPA and DHA further enhanced Bax expression. DG decreased glutathione content, increased reactive oxygen species (ROS) and oxidized glutathione (GSSG) production, the intake of EPA or DHA caused greater ROS and GSSG formation. DG treatments up-regulated the protein expression of p47phox and gp91phox, and the intake of EPA or DHA led to greater p47phox and gp91phox expression. DG increased brain prostaglandin E2 (PGE2) levels, and cyclooxygenase (COX)-2 expression and activity, the intake of EPA or DHA reduced brain COX-2 activity and PGE2 formation. DG enhanced brain p53, p16 and p21 expression. EPA and DHA intake led to greater p21 expression, and EPA only caused greater p53 and p16 expression. Conclusion: These findings suggest that these two PUFAs have toxic effects toward aging brain.

Mechanism of ginsenoside Rg1 renal protection in a mouse model of d-galactose-induced subacute damage

Context Ginseng is a widely used herbal medicine in China but its mechanism of action remains unclear.Objective The objectives of this work were to study the protective effect of ginsenoside Rg1 on subacute murine renal damage induced by d-galactose and its mechanism.Materials and methods C57BL/6J mice were injected with 120 mg/kg/d (sc) d-galactose for 1 week, followed by a combined treatment of Rg1 20 mg/kg/d (ip) and 120 mg/kg/d d-galactose (sc) for 5 weeks. Mice were injected with the 0.9% saline 0.2 mL/d (sc) and 120 mg/kg/d d-galactose (sc) for 6 weeks in the control group and the d-galactose group, respectively. After 6 weeks, urea, creatinine, uric acid, cystatin (Cys-C), senescence-associated β-galactosidase (SA-β-gal) staining positive kidney cells, superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), glycation end products (AGEs) and 8-hydroxy-2 deoxyguanosine (8-OH-dG) were measured.Results Treatment with Rg1 ameliorated kidney function and aging state (urea from 17.19 ± 1.09 to 15.77 ± 1.22 mmol·L − 1, creatinine from 29.40 ± 5.72 to 22.60 ± 3.97 μmol·L − 1, uric acid from 86.80 ± 5.97 to 72.80 ± 10.61 μmol·L − 1, Cys-C from 0.23 ± 0.03 to 0.18 ± 0.05 mg·L − 1, ROD of SA-β-gal from 56.32 ± 10.48 to 26.78 ± 7.34, SOD from 150.22 ± 19.07 to 190.56 ± 15.83 U·(mg·prot) −1, MDA from 9.28 ± 1.59 to 3.17 ± 0.82 nmol·(mg·prot) −1, GSH-PX from 15.68 ± 2.11 to 20.32 ± 2.96 U·(mg·prot) −1 as well as regulated glomerulus morphology (glomerulus diameter from 775.77 ± 18.41 to 695.04 ± 14.61 μm, renal capsule width from 39.56 ± 3.51 to 31.42 ± 2.70 μm, glomerulus basement membrane from 206.03 ± 16.22 to 157.27 ± 15.70 nm, podocyte slit from 55.21 ± 8.55 to 37.63 ± 6.65 nm).Conclusions Ginsenoside Rg1 can antagonise d-galactose subacute renal damage in mice and this may occur due to alleviating oxidative stress injury.

EFFECTS OF POLYSACCHARIDES FROM YULANGSAN ON D-GALACTOSE INDUCED AGING MODEL MICE

Background: Yulangsan (Millettia pulchra Kurz ) is a traditional herb that is often used in popular folk medicine in the Guangxi Zhuang Autonomous Region of China. Previous studies had showed that polysaccharides extracted from Yulangsan (YLSPS) has strong anti -ROS activities. However, its anti-aging effect has not yet been studied. Objective: In the present study, we study the effects of the YLSPS on aging mice induced by D-galactose. Method: Six week old male Kunming (18-22g) 40 mice were randomly divided into 4 groups: Normal, Model, Model treated with YLSPS and Model treated with vitamin E groups, 10 mice in each group. The mice were artificially induced aging by subcutaneously injection with D-galactose 200mg/kg/day for 8 weeks. At the same time, YLSPS 600mg/kg/day or Vit E 200mg/kg/day were simultaneously treated to them once daily by intragastric gavage. Activities of ROS-scavenging enzymes, like superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC) and catalase (CAT), as well as levels of Malondialdehyde (MDA) in liver, brain, and serum were determined respectively. IL2, IL6, and AGEs in serum were detected by ELISA assay and the protein level of p21 and p53 were measured by Western blotting. Results: YLSPS could increased thymus index and spleen index, improve SOD, CAT, GSHPx, T-AOC activities, reduce MDA level compared with aging mice induced by D-galactose group. Conclusion: All of these findings that YLSPS may have effect to suppress the aging process through enhancing antioxidant activity and scavenging free radical. Key words: Polysaccharide; Millettia pulchra; anti-aging; D-galactose.

Neuronal marker recovery after Simvastatin treatment in dementia in the rat brain: In vivo magnetic resonance study

The aim of study was to search for new biomarkers with a magnetic resonance technique to identify the early stages of dementia, induced by d-galactose, and evaluate Simvastatin therapy. Localized proton magnetic resonance spectroscopy measurements showed a significant decrease in the concentration of N-acetylaspartate+N-acetylaspartylglutamate and myo-inositol in the d-galactose group compared to the control group, and, conversely, an increase of N-acetylaspartate+N-acetylaspartylglutamate in the d-galactose/Simvastatin group. Using a saturation transfer experiment, with phosphorus magnetic resonance spectroscopy, we observed a significant elevation of the forward rate constant of the creatine kinase reaction in the brains of the d-galactose group compared to controls, and subsequently, a significant reduction of this reaction in the d-galactose/Simvastatin group. Spatial learning and memory were evaluated using the modified Morris water maze test. The dynamics of the learning process represented by the learning index revealed a significant reduction in learning in the d-galactose group, but the deficits as a consequence of the d-galactose effects were recovered in the d-galactose/Simvastatin group, in which the learning dynamics resembled those of the control group. By determining the thiobarbituric acid reactive substances and total coenzyme Q9 in plasma, we have shown that long-term administration of d-galactose created conditions for oxidative stress, and that the administration of Simvastatin decreased oxidative stress in plasma. Volumetry analyses from the hippocampal area show a reduction in the segmented area in the d-galactose group, compared with the control group, and an enlarged area in the hippocampus in the d-galactose/Simvastatin group.

Effect of Capparis spinosa L. on cognitive impairment induced by D-galactose in mice via inhibition of oxidative stress.

To determine the phenolic acid levels and DNA damage protection potential of Capparis spinosa L. seed extract and to investigate the effect of the extract on cognitive impairment and oxidative stress in an Alzheimer disease mice model. Thirty BALB/c mice divided into 5 groups (control, D-galactose, D-galactose + C. spinosa 50, D-galactose + C. spinosa 100, D-galactose + C. spinosa 200) were used. Mice were administered an injection of D-galactose (100 mg/kg, subcutaneous) and orally administered C. spinosa (50, 100, or 200 mg/kg) daily for 8 weeks. Syringic acid was detected and the total amount was 204.629 µg/g. Addition of 0.05 mg/mL C. spinosa extract provided significant protection against the damage of DNA bands. C. spinosa attenuated D-galactose-induced learning dysfunctions in mice and significantly increased memory retention. Malondialdehyde (MDA) levels increased and superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities decreased in the D-galactose group. C. spinosa (200 mg/kg body weight) significantly decreased MDA level and increased SOD, GPx, and CAT activities. These results show that C. spinosa has the potential in ameliorating cognitive deficits induced by D-galactose in mice and the antioxidant activity may partially account for the improvement of learning and memory function.

Calorie Restriction Down-Regulates Expression of the Iron Regulatory Hormone Hepcidin in Normal and d-Galactose–Induced Aging Mouse Brain

It has been shown that iron progressively accumulates in the brain with age. Calorie restriction (CR) may allay many of the adverse effects of aging on the brain, yet the underlying mechanisms, in particular in relation to brain iron metabolism, remain unclear. This study aimed to investigate the role of CR in the regulation of cerebral cellular iron homeostasis. C57BL/6 mice were randomly divided into four groups of eight. The control group was fed a conventional diet ad libitum; the CR group received 70% of the calories of the control mouse intake per day; the D-galactose (D-gal) group received subcutaneous injection of D-gal at a dose of 100 mg/kg once daily to produce mouse model of aging; the D-gal plus CR group received both of the two interventions for 14 weeks. The Morris water maze (MWM) was employed to test the cognitive performance of all animals, and the expression of iron regulatory genes, ferroportin and hepcidin, in the cortex and hippocampus were detected by quantitative real-time PCR. Compared to the controls, the D-gal group mice showed significant spatial reference memory deficits in the MWM test, whereas the D-gal-CR group mice exhibited almost normal cognitive function, indicating that CR protects against D-gal-induced learning and memory impairment. Hepcidin mRNA expression was increased in the D-gal group, decreased in the CR group, and was basically unchanged in the D-gal-CR group. There was no statistical difference in the transmembrane iron exporter ferroportin expression between control and any of the experimental groups. The results suggest that the anti-aging effects of CR might partially lie in its capacity to reduce or avoid age-related iron accumulation in the brain through down-regulating expression of brain hepcidin--the key negative regulator for intracellular iron efflux--and that facilitating the balance of brain iron metabolism may be a promising anti-aging measure.