Young Control Rats Research Articles

A Water-soluble Form of Dihydroquercetin Reduces LPS-induced Astrogliosis, Vascular Remodeling, and mRNA VEGF-A Levels in the Substantia Nigra of Aged Rats

Background: The age-dependent sporadic form of PD is characterized by the degeneration of dopaminergic (DA) neurons in the Substantia Nigra (SN), gliosis, and vascular changes. Vascular changes may contribute to the onset of the disease and exacerbate the neurodegenerative process, as some vascular changes occur before the onset of neuronal loss. To demonstrate the anti-neuroinflammatory efficacy of a new compound, a water-soluble form of dihydroquercetin (DHQ-WF), we studied the structural changes of microcirculatory vasculature, astroglial GFAP, and vascular endothelial growth factor –A (VEGF-A) mRNA expression in the SN of young and old rats after unilateral nigral treatment by lipopolysaccharide (LPS) and oral administration of DHQ-WF. Materials and methods: The experiments were performed on 18 young (8 weeks - 10 weeks old; 250 g - 320 g) and 18 old (18 months - 19 months old; 390 g - 450 g) male Vistar rats. Young and adult rats from the experimental groups were stereotactically injected with 2 μL LPS solution (LPS from Escherichia coli; 0,01 μL/mL) into one side of the SN. Control young and old rats were similarly injected with 2 μL sterile saline. Half of the animals in both the control and experimental groups (6 animals in each group) received a 2 ml solution containing DHQ-WF at a concentration of 3 mg/ml orally every day. After 8 weeks, brains were harvested and serial cryostat sections were prepared for histochemical (FITC-labeled tomato lectin), immunohistochemical (anti-GFAP Antibody, Cy3 Conjugate) staining, and real-time PCR (mRNA VEGF-A). Results: Eight weeks after LPS injection into the SN, a significant excess of areas occupied by cell bodies and processes of astroglial cells, the density of microcirculatory vessels, and mRNA VEGF-A expression was observed in old animals compared to control old animals and young LPS-treated rats. Oral administration of DHQ-WF to LPS-treated rats resulted in a significant reduction of these parameters in old animals. Conclusion: Injection of LPS into rat SN induces neuroinflammation and vascular angiogenesis, maximally expressed in old animals. Administration of DHQ-WF for 8 weeks significantly reduces these LPS-induced changes. DHQ-WF may be an effective treatment for reducing the effects of neuroinflammation in the aging brain.

The role of cellular senescence in profibrillatory atrial remodelling associated with cardiac pathology.

Cellular senescence is a stress-related or aging response believed to contribute to many cardiac conditions; however, its role in atrial fibrillation (AF) is unknown. Age is the single most important determinant of the risk of AF. The present study was designed to (i) evaluate AF susceptibility and senescence marker expression in rat models of aging and myocardial infarction (MI), (ii) study the effect of reducing senescent-cell burden with senolytic therapy on the atrial substrate in MI rats, and (iii) assess senescence markers in human atrial tissue as a function of age and the presence of AF. AF susceptibility was studied with programmed electrical stimulation. Gene and protein expression was evaluated by immunoblot or immunofluorescence (protein) and digital polymerase chain reaction (PCR) or reverse transcriptase quantitative PCR (messenger RNA). A previously validated senolytic combination, dasatinib and quercetin, (D+Q; or corresponding vehicle) was administered from the time of sham or MI surgery through 28 days later. Experiments were performed blinded to treatment assignment. Burst pacing-induced AF was seen in 100% of aged (18-month old) rats, 87.5% of young MI rats, and 10% of young control (3-month old) rats (P ≤ 0.001 vs. each). Conduction velocity was slower in aged [both left atrium (LA) and right atrium (RA)] and young MI (LA) rats vs. young control rats (P ≤ 0.001 vs. each). Atrial fibrosis was greater in aged (LA and RA) and young MI (LA) vs. young control rats (P < 0.05 for each). Senolytic therapy reduced AF inducibility in MI rats (from 8/9 rats, 89% in MI vehicle, to 0/9 rats, 0% in MI D + Q, P < 0.001) and attenuated LA fibrosis. Double staining suggested that D + Q acts by clearing senescent myofibroblasts and endothelial cells. In human atria, senescence markers were upregulated in older (≥70 years) and long-standing AF patients vs. individuals ≤60 and sinus rhythm controls, respectively. Our results point to a potentially significant role of cellular senescence in AF pathophysiology. Modulating cell senescence might provide a basis for novel therapeutic approaches to AF.

Carvacrol improved learning and memory and attenuated the brain tissue oxidative damage in aged male rats

Introduction: Aging is an unavoidable process in the body that is accompanied by impaired tissue homeostasis and various changes. Carvacrol has attracted considerable attention for its wide range of pharmacological activities. Therefore, this study attempted to explore the protective effect of carvacrol in aged rats.Materiel and methods: The aged rats were given carvacrol (15 or 30 mg/kg/day) for 4 weeks. Morris water maze and passive avoidance tests were used to determine the learning and memory abilities of the rats. The hippocampus and cortex samples were taken for biochemical analysis.Results: In comparison to young control rats, aged control rats showed learning and memory deficits. There was improvement in the Morris water navigation test and passive avoidance test performance in the treatment groups versus the aged control group. An increment in malondialdehyde (MDA) and a decrease in total thiol groups in the hippocampus and cortex samples of aged control rats in comparison to the young control group were observed. Carvacrol decreased MDA levels and increased total thiol groups in the hippocampus and cortex samples of aged rats.Conclusion: Carvacrol improved learning and memory in aged rats, probably through its anti-oxidation effects.

Intake of Fluted Pumpkin Seeds Rebalances Oxidative Stress Parameters in the Aged Rat’s Testes

The effect of fluted pumpkin seeds (FPS) consumption on the antioxidant status of the testes of aged Wistar rats was evaluated in this study. Sixty (50 aged, 6 months old, and 10 young, 2 months old) rats were divided into six groups of 10 per each group. Testosterone (15 mg kg-1 body weight, once weekly for 40 days) was injected intraperitoneally and used as positive control. FPS intake (50, 100, and 200 mg kg-1 body weight) or vehicle control (corn oil) were administered orally, twice weekly for 40 days and compared with the untreated aged and young control rats. Changes in antioxidant status in the testis of the aged rats was reflected as increased superoxide dismutase and catalase activities and glutathione and decreased lipid peroxidation levels which were attenuated more efficiently by the lowest dose FPS (50 mg kg-1 body weight). Additionally, nitrite concentration that was found to be diminished in the aged rats was raised to the young control values after intake of the FPS (50 mg kg-1 body weight). As expected, testosterone injection increased endogenous testosterone concentration and also remained higher in the untreated aged animals than in young control and treated aged rats. In conclusion, compromised antioxidant defense system of the testes that is associated with ageing could be reversed to the status of the young control by the intake of FPS.

STUDY OF FLAVONOIDS IN CAMEL THORN AND THEIR ACTIVITY IN ANGIOTENSIN-CONVERTING ENZYME REDUCTION

The article is dedicated to the study of polyphenolic compounds in extracts of camel thorn (Alhagi pseudoalhagi). Using UV spectroscopy and HPLC-MS/MS, the composition and content of flavonoids in camel thorn extracts were determined. Characteristic absorption bands for flavonoids were identified through optical spectroscopy. Data on the substance composition in the extract were obtained using the HPLC-MS/MS method. Semi-quantitative analysis of flavonoids was performed using the internal standard addition method with rutin. More than twenty flavonoids were detected in the 70% ethanolic extract. Rhamnetin 3-neohesperidoside and Isorhamnetin 3-neohesperidoside were the major components. The results could be applied in the production of antioxidant pharmaceuticals in the pharmaceutical industry. Comparison of the effects of camel thorn extract with the flavonoid taxifolin showed that the extract is equally effective in preventing early stages of atherosclerosis in the aorta caused by increased angiotensin-converting enzyme activity. At an extract concentration of 0.2%, the activity of the angiotensin converting enzyme in elderly rats and an imals treated with a NO-synthase inhibitor decreases to the values of young control rats.

Examining the role of Drp1 in age‐related microvascular dysfunction

IntroductionAdvanced age is a well‐documented risk factor for coronary microvascular dysfunction. Recent evidence has underscored that advanced age is associated with a shift toward greater mitochondrial fission rather than mitochondrial fusion. Therefore, understanding key regulators of the balance of mitochondrial fission/fusion may help to identify the contribution of mitochondrial dynamics on microvascular endothelial function. DRP1 is a pro‐fission factor that has been linked to several disease manifestations including advanced age and cardiovascular disease (CVD).MethodsCoronary arteries were obtained from young control rats (YC, 15‐20 weeks) and old control rats (OC, 95‐100 weeks) and expression of DRP1 was assessed by immunofluorescence (IF). A novel rat model crossing flox‐stop DRP1‐GFP rats and VECAD‐Cre expressing rats was generated to assess the vascular and cardiac effects of endothelial‐specific DRP1 overexpression. Endothelial vascular function was determined by endothelial‐dependent flow‐mediated dilation while cardiac function was determined by echocardiography at 20 weeks old and compared to wildtype control group.ResultsData from IF comparing coronary arteries from YC and OC show that OC had higher DRP1 expression compared to YC (1.42 to 1.00, p=0.01). Max dilation measurements, taken at 20‐weeks of age, showed that DRP1‐overexpressing rats was decreased when compared to wildtype (WT) control rats (31.4 to 60.1, p=0.009). Echocardiography revealed that DRP1‐overexpressing rats showed a trend towards a decrease in coronary flow reserve and increased HR in response to a dobutamine stressor when compared to WT. Ejection fraction between the two groups remained largely preserved.ConclusionOur data demonstrate that there may be a critical role for DRP1 within in vitro vasodilation that mimics a CVD pro‐fission phenotype. Future studies will be performed to further evaluate the role of DRP1 in response to other stressors well‐established in age‐dependent CVD such as hypertension.

The Effect of Tauroursodeoxycholic Acid (TUDCA) Treatment on Pregnancy Outcomes and Uterine Artery Function in a Rat Model of Advanced Maternal Age

IntroductionAdvanced maternal age (AMA; ≥35 years) increases the risk of pregnancy complications and adverse pregnancy outcomes such as fetal growth restriction. Endoplasmic reticulum (ER) and oxidative stress have been linked to vascular dysfunction (i.e. reduced nitric oxide [NO] &amp; increased endothelin‐1 [ET‐1] levels) in aging. Tauroursodeoxycholic acid (TUDCA) is an ER and oxidative stress inhibitor and has been shown to improve embryo development in vitro. However, whether treatment with TUDCA can help improve uterine vascular function and pregnancy outcomes in AMA has not been investigated. We hypothesize that TUDCA treatment will improve pregnancy outcomes and vascular function in a rat model of AMA.MethodsYoung (4 months) and aged (9.5 months of age; ~35 years in humans) pregnant control or TUDCA‐treated rats were studied on gestational day 20 (term=22 days). Following euthanasia, pregnancy outcomes (litter size, resorption sites, fetal weight, and placental weight) were recorded, and main uterine arteries were isolated. Endothelium‐dependent vasorelaxation to methacholine (MCh) was assessed by wire myography in the presence/absence of pan NO synthase inhibitor (L‐NAME). In addition, big‐endothelin‐1 (bET‐1) vasoconstriction responses were evaluated, in the presence/absence of endothelin converting enzyme (ECE) inhibitor (CGS). Data summarized as maximum vasodilation response (Emax) and analyzed by two‐way ANOVA with Sidak’s post‐test, p&lt;0.05 was considered significant.ResultsFetal body weights were reduced in AMA control rats compared to young dams (p&lt;0.017), while TUDCA treatment tended to increase fetal body weight in AMA rats (p=0.067); TUDCA did not affect fetal body weight in the young dams. Placental weights, litter size and number of resorptions were similar in all the groups. The maximum relaxation (Emax) to MCh was impaired in the AMA dams compared to young dam (p=0.047), and this reduced relaxation was not evident in the TUDCA treated AMA dams. Ex vivo treatment with L‐ NAME showed an increased contribution of NO in young TUDCA treated rats (ΔEmax [differences in Emax]; p=0.043) compared to young control rats but not in AMA rats. Ex vivo treatment with CGS demonstrated reduced ECE conversion in young TUDCA treated rats (Emax; p&lt;0.001), while CGS had no effect in the young control dams or both AMA groups.ConclusionOur study assessed TUDCA as an intervention to improve the pregnancy outcomes in a rat model of AMA. An increased fetal body weight suggests a beneficial effect of TUDCA in AMA pregnancies. However, contrary to our hypothesis, in young dams, TUDCA increased NO contribution and reduced ECE effect, but did not affect uterine artery function in AMA dams. Nevertheless, TUDCA has the potential to improve pregnancy outcomes in complicated pregnancies, but further studies are warranted.

Moderate Aerobic Training Inhibits Middle-Aged Induced Cardiac Calcineurin-NFAT Signaling by Improving TGF-ß, NPR-A, SERCA2, and TRPC6 in Wistar Rats.

ObjectiveThe purpose of this study was to investigate the effect of moderate-intensity training on the calcineurin/ nuclear factor of activated t-cells (NFAT) pathway and factors affecting it in the middle-age Wistar rats.Materials and MethodsIn this experimental study, 40 young (n=10, 4-month-old) and middle-aged (n=30, 13-15 months old) Wistar rats were included in this experimental study. All young and 10 middle-aged rats did not training and served as a control comparision; while the remaining 20 middle-aged rats were trained at moderate intensity for 4-weeks (n=10) or 8-weeks (n=10) on a treadmill (speed: 16 m/minutes, slope: 0%, distance: 830 m, duration: 54 minutes). Results Calcineurin tissue expression was increased in the middle-aged control rats compared to the young control rats (P=0.001). Expression of sarco/endoplasmic reticulum Ca2+ -ATPase (SERC2A), natriuretic peptide receptor-A (NPR-A), phospholamban (PLB), plasma membrane Ca2+ ATPase (PMCA4b), and p-AKT was significantly decreased in the heart tissue of middle-aged control compared to the young control rats (P=0.001). Furthermore, transforming growth factor beta (TGF-β), including transient receptor potential canonical 6 (TRPC6), were up-regulated in the heart tissue of middle-aged control compared to the young control rats (P=0.001). However, aerobic training inhibited this pathway and reversed all changes in the trained middle-aged rats. ConclusionAerobic training effectively inhibited the calcineurin/NFATc pathway and modulated intracellular Ca2+ levels at least partially by restoring NPR-A, SERCA2, p-PLB, and p-AKT, and decreasing TRPC6 and TGF-β levels.

Advanced Glycation End Products Increase Salivary Gland Hypofunction in d-Galactose-Induced Aging Rats and Its Prevention by Physical Exercise.

A declined salivary gland function is commonly observed in elderly people. Advanced glycation end products (AGEs) are believed to contribute to the pathogenesis of aging. Although physical exercise is shown to increase various organ functions in human and experimental models, it is not known whether it has a similar effect in the salivary glands. In the present study, we evaluated the AGEs burden in the salivary gland in the aging process and the protective effect of physical exercise on age-related salivary hypofunction. To accelerate the aging process, rats were peritoneally injected with D-galactose for 6 weeks. Young control rats and d-galactose-induced aging rats in the old group were not exercised. The rats in the physical exercise group ran on a treadmill (12 m/min, 60 min/day, 3 days/week for 6 weeks). The results showed that the salivary flow rate and total protein levels in the saliva of the d-galactose-induced aging rats were reduced compared to those of the young control rats. Circulating AGEs in serum and secreted AGEs in saliva increased with d-galactose-induced aging. AGEs also accumulated in the salivary glands of these aging rats. The salivary gland of aging rats showed increased reactive oxygen species (ROS) generation, loss of acinar cells, and apoptosis compared to young control mice. However, physical exercise suppressed all of these age-related salivary changes. Overall, physical exercise could provide a beneficial option for age-related salivary hypofunction.

The effects of long-term moderate exercise and Western-type diet on oxidative/nitrosative stress, serum lipids and cytokines in female Sprague Dawley rats

PurposeRegular exercise reduces obesity and the risk of cardiovascular disease. However, health-promoting benefits of physical activity are commonly associated with increased inflammation and oxidative stress. Here, we tested whether constant moderate exercise is able to prevent or attenuate the oxidative/nitrosative stress, inflammation, and serum lipids in lean and obese rats.MethodsFour-month-old female Sprague Dawley rats received standard or a high-fat diet. Animals were subjected to a physical activity protocol, consisting of 30 min forced treadmill exercise for 5 consecutive days per week during 10 months. Baseline and sedentary (non-exercised) rats were used as controls. Lipids, oxidized low-density lipoprotein cholesterol, nitric oxide metabolites, and pro- and anti-inflammatory markers were measured in blood collected upon euthanasia.ResultsAt variance to young baseline control rats, 14-month-old animals fed normal diet had increased plasma lipid levels, including total cholesterol and triglycerides, which were further elevated in rats that consumed a high-fat diet. While treadmill exercise did not lower the amount of serum lipids in standard diet group, forced physical activity reduced non-high-density lipoprotein cholesterol in response to high-fat diet feeding. Exercised rats fed standard diet or high-fat diet had lower abundancy of nitric oxide metabolites, which coincided with increased levels of oxidized low-density lipoprotein cholesterol. Accordingly, the amount of nitric oxide metabolites correlated inversely with oxidized low-density lipoprotein cholesterol and homo-arginine. Exercise significantly reduced inflammatory cytokines in high-fat diet fed rats only.ConclusionOur study suggests that regular exercise alters the equilibrium between oxidative and anti-oxidative compounds and reduces pro-inflammatory cytokines.

Effect of Wheatgrass (Triticum aestivum Linn) Extracts as Source of Antioxidants on some biochemical parameters and Markers of Aging

Aging is defined of the process of getting older, or an artificial process to make someone or something look older and loss of function. There has been a global trend toward using of natural phytochemicals present in natural sources as antioxidants and functional foods. Wheatgrass has been proved to contain essential amino acids, active enzymes, flavonoids, chlorophyll, minerals, proteins and vitamins This study was aimed to see the effect of ethanolic and water extracts of wheat grass as natural sources of antioxidants on markers of aging in rats. 36 Sprague-Dawley female rats were obtained. six rats weighing 112 to 125 g considered as control young and therefore the rest of rats weighing 256 to 267 g considered as aged rats their age ranges between 18 month and to 24 month. The rats were divided into 5 groups, the primary and second groups consumed basal diet and thought of as control young and control aged rats, respectively, while the opposite four groups administered ethanolic and water extracts of Wheatgrass powder at doses of 10 and 20 mg for every rat once daily. Blood samples were taken from rats and examined for markers of aging as, the activity of SOD and lipid peroxidation (MDA), total cholesterol, HDL, LDL, VLDL, triglycerides levels and liver functions. Aging resulted during a significant deterioration altogether tested markers of aging including, SOD and MDA, total protein, globulin, albumin HDL-cholesterol. Also, a big increase in, ALT, AST triglycerides, total cholesterol, LDL and VLDL levels was observed during aging. Administering aged rats with Wheatgrass ethanolic and water extracts caused a big improvement within the above markers and returned the abnormal markers back to the traditional levels Approximately as found on top of things young rats especially at 20 mg from wheatgrass. So, it might be recommended that wheatgrass ethanolic extracts (70) might be wont to improve the abnormal changes in health state of aging human and return them to just about the traditional state found in younger human by employing a natural source of antioxidants like Wheatgrass.

Effect of Age and Sodium Alendronate on Femoral Fracture Repair: Biochemical and Biomechanical Study in Rats.

BackgroundBisphosphonates are drugs widely used to reduce bone resorption, increase bone mineral density and control age-related bone loss. Although there are studies reporting differences in bone structure between young and old adults, it is still difficult to predict changes related to bone aging. The aim of this study was to evaluate the effect of age and sodium alendronate on bone repair of femoral fractures in rats.MethodsWistar rats (n = 40) were allocated into groups: O (control old-rats), Y (control young-rats), OA (alendronate old-rats) and YA (alendronate young-rats). All animals underwent linear fracture surgery followed by fixation. Groups OA and YA received 1 mg/kg alendronate three times a week until euthanasia. Biochemical analysis of calcium and alkaline phosphatase was done. After euthanasia, femurs were evaluated in relation to cross-section and flexural strength, with three-point bending test. Data were submitted to statistical analysis with significance level of 0.05.ResultsThere was no difference in calcium and alkaline phosphatase levels (p > 0.05). Young animals presented lower cross-section than older animals (p < 0.05). Only fractured side, young animals presented major flexural strength than older animals (p < 0.05). There was no difference between the animals that used or not alendronate in relation to cross-section and flexural strength (p > 0.05). When compared fractured and non-fractured femurs, major cross-section on fractured side was observed (p < 0.05). Flexural strength presented higher values in femurs on non-fractured side (p < 0.05). There was correlation of weight and cross-section (R = +0.91) and weight with flexural strength of fractured and non-fractured side, respectively (R = −0.97 and −0.71).ConclusionIn short, there was no difference of calcium and alkaline phosphatase during the bone repair process. Age has influence in cross-section and flexural strength. Alendronate showed no association with these factors.

Effects of Chronic Photobiomodulation with Transcranial Near-Infrared Laser on Brain Metabolomics of Young and Aged Rats.

Since laser photobiomodulation has been found to enhance brain energy metabolism and cognition, we conducted the first metabolomics study to systematically analyze the metabolites modified by brain photobiomodulation. Aging is often accompanied by cognitive decline and susceptibility to neurodegeneration, including deficits in brain energy metabolism and increased susceptibility of nerve cells to oxidative stress. Changes in oxidative stress and energetic homeostasis increase neuronal vulnerability, as observed in diseases related to brain aging. We evaluated and compared the cortical and hippocampal metabolic pathways of young (4months old) and aged (20months old) control rats with those of rats exposed to transcranial near-infrared laser over 58 consecutive days. Statistical analyses of the brain metabolomics data indicated that chronic transcranial photobiomodulation (1) significantly enhances the metabolic pathways of young rats, particularly for excitatory neurotransmission and oxidative metabolism, and (2) restores the altered metabolic pathways of aged rats towards levels found in younger rats, mainly in the cerebral cortex. These novel metabolomics findings may help complement other laser-induced neurocognitive, neuroprotective, anti-inflammatory, and antioxidant effects described in the literature.

Chrysin's Impact on Oxidative and Inflammation Damages in the Liver of Aged Male Rats.

The purpose of this research was to investigate the effect of chrysin on one of the natural antioxidants on aging progression in an animal model. Oxidative stress and inflammation increase in hepatic tissue during aging, leading to liver dysfunction. The current research was conducted to show the effect of chrysin on the activities of antioxidant enzyme (catalase, glutathione peroxidase, and superoxide dismutase), serum nitric oxide (NO), and lipid peroxidation as well as inflammatory cytokines (TNF-α, IL-6, and IL-1β) of aging rats. Male Wistar rats of different ages, 2, 10, and 20 months, were randomly divided into six groups as follows (n=8, per each group): young control rats (C2), young CH-treated rats (CH2), middle- aged control rats (C10), middle-aged CH-treated group (CH10), aged control group (C20), and aged CH-treated group (CH20). Chrysin (20 mg/kg) was administered intraperitoneally once a day for 30 days. Present findings indicated that chrysin treatment ameliorated the increased liver levels of lipid peroxidation, TNF-α, and IL-1β as well as serum levels of NO. The findings suggest that chrysin could be effective against the progression of ageinduced damage by modulation of oxidant-antioxidant system and inflammatory response.

The age-dependent effect of pre-pubertal castration on anxiety-like behaviour in male rats.

Adolescence is considered to be a critical period of sex hormone action (re)organising the brain and determining the behavioural phenotype. Such organisational effects in the brain might be the cause of sex differences in some behavioural features. In this experiment, we aimed to examine the role of pubertal sex hormones in development of anxiety in male rats. Male rats underwent gonadectomy prior to puberty onset, and were tested for explorative and anxiety-like behaviour in adolescence as well as in young adulthood. In adolescence, but not in adulthood, gonadectomised rats spend by 50% more time (p<.05) in the centre zone of the open-field than sham-operated counterparts. Young adult gonadectomised rats showed approximately 1.5-fold greater exploratory activity, in both open field (p<.001) and elevated plus maze (p<.01), in comparison with young adult control rats. Our results indicate that pre-pubertal castration may have test-specific anxiolytic effect in adolescent male rats, and it may attenuate the decline in explorative behaviour in young adult males. These differences in short- and long-term effects of gonadectomy could explain some contradictory results of previous studies on the role of testosterone in anxiety-like behaviour of male rodents. Thus, the age-specific consequences of pre-pubertal hormone deprivation should be considered.

Intermedin1-53 attenuates aging-associated vascular calcification in rats by upregulating sirtuin 1.

Vascular calcification is a common phenomenon in older adults. Intermedin (IMD) is a cardiovascular bioactive peptide inhibiting vascular calcification. In this study, we aimed to investigate whether IMD1-53 attenuates aging-associated vascular calcification. Vascular calcification was induced by vitamin D3 plus nicotine (VDN) in young and old rats. The calcification in aortas was more severe in old rats treated with VDN than young control rats, and IMD expression was lower. Exogenous administration of IMD1-53 significantly inhibited the calcium deposition in aortas and the osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs) in VDN-treated old rats. Moreover, levels of aging-related p16, p21 and β-galactosidase were all greatly decreased by IMD1-53. These results were further confirmed in rat and human VSMCs in vitro. In addition, IMD-deficient mouse VSMCs showed senescence features coinciding with osteogenic transition as compared with wild-type mouse VSMCs. Mechanistically, IMD1-53 significantly increased the expression of the anti-aging factor sirtuin 1 (sirt1); the inhibitory effects of IMD1-53 on calcification and senescence were blocked by sirt1 knockdown. Furthermore, preincubation with inhibitors of PI3K, AMPK or PKA efficiently blunted the upregulatory effect of IMD1-53 on sirt1. Consequently, IMD1-53 could attenuate aging-associated vascular calcification by upregulating sirt1 via activating PI3K/Akt, AMPK and cAMP/PKA signaling.

Effects of nutraceutical intervention on serum proteins in aged rats

Aging is associated with many pathophysiological changes that could lead to the onset of degenerative disease. Some of the physiological changes that occur with aging include increased inflammation and decreased stem cell proliferation, leading to decreased capacity for tissue regeneration and loss of function. In previous studies, we and others have found nutraceutical intervention to ameliorate some of the deleterious effects associated with aging. In particular, we have previously shown that NT-020, a supplement composed of a proprietary blend of blueberries, green tea, vitamin D3, and carnosine, is able to rescue age-related cognitive deficits, impaired neurogenesis, and inflammation in rats. We have also previously demonstrated that stem cells cultured with old serum showed decreased proliferation; however, when stem cells were cultured in serum from old rats given a diet supplemented with NT-020, proliferation did not differ from that of cells cultured with serum from young rats. While it is clear that NT-020 is exerting a therapeutic, anti-aging effect, the mechanisms of action were yet to be fully elucidated.To that end, in the present study, we conducted a bioinformatics experiment to examine the rat proteome of serum from young and old control rats and young and old rats given a diet supplemented with NT-020. Serum from old rats showed an increase in some inflammatory and pro-aging factors while serum from old rats given a diet supplemented with NT-020 showed an increase in some anti-aging factors, most notably proteins associated with the complement system and autophagy. A number of immune functions that increase with age were shown to be downregulated with NT-020 treatment.

Impact of intermittent voluntary ethanol consumption during adolescence on the expression of endocannabinoid system and neuroinflammatory mediators

The adolescent brain displays high vulnerability to the deleterious effects of ethanol, including greater risk of developing alcohol use disorder later in life. Here, we characterized the gene expression of the endocannabinoid system (ECS) and relevant signaling systems associated with neuroinflammation and emotional behaviors in the brain of young adult control and ethanol-exposed (EtOH) rats. We measured mRNA levels of candidate genes using quantitative real time PCR in the medial prefrontal cortex (mPFC), amygdala and hippocampus. EtOH rats were generated by maintenance on an intermittent and voluntary ethanol consumption during adolescence using the two-bottle choice paradigm (4 days/week for 4 weeks) followed by 2 week-withdrawal, a time-point of withdrawal with no physical symptoms. Mean differences and effect sizes were calculated using t-test and Cohen's d values. In the mPFC and hippocampus, EtOH rats had significantly higher mRNA expression of endocannabinoid-signaling (mPFC: Ppara, Dagla, Daglb and Napepld; and hippocampus: Cnr2, Dagla and Mgll) and neuroinflammation-associated genes (mPFC: Gfap; and hippocampus: Aif1) than in controls. Moreover, EtOH rats had significantly higher mRNA expression of neuropeptide Y receptor genes (Npy1r, Npy2r and Npy5r) in the hippocampus. Finally, EtOH rats also displayed higher plasma endocannabinoid levels than controls. In conclusion, these results suggest that adolescent ethanol exposure can lead to long-term alterations in the gene expression of the ECS and other signaling systems involved in neuroinflammation and regulation of emotional behaviors in key brain areas for the development of addiction.

Withaferin-A Protects the Nigral Dopamine Neuron and Recovers Motor Activity in Aged Rats

Withaferin-A (WA) was evaluated for its neuroprotective efficacy on the dopamine (DA) neurons of the substantia nigra (SN) and striatum (ST) in aged rats. Wistar albino rats were divided into group I, young (3 months old); group II, aged (24 months old); group III, aged rats supplemented with WA (50 mg/kg bodyweight once per day for 30 days), and group IV, young rats supplemented with WA (50 mg/kg bodyweight). At the end of the experiment period, the animals were subjected to various motor behavior analyses, and were sacrificed by transcardial perfusion. The brains were dissected out and subjected to various analyses, including histological, histomorphometrical, and immunolocalization of the tyrosine hydroxylase (TH) enzyme. The data of rotarod analysis (p < 0.001) showed a significant motor impairment in aged rats (number of falls 10.2 ± 0.86) and reduction in retention time (31.23 ± 2.56 s) compared to young controls (2.41 ± 0.35 and 84.05 ± 5.15 s). The stride length was significantly reduced (p < 0.001) in aged rats (4.21 ± 0.57 and 4.38 ± 0.61 cm) when compared to young control rats (6.98 ± 0.25 and 7.13 ± 0.70 cm). The histomorphometric data of the aged animals showed a significant reduction in the neuronal diameter (p < 0.001), density (p < 0.001), and volume (p < 0.001) in the SN of aged rats when compared to young rats. Immunohistology demonstrated a marked reduction in the levels of TH enzyme in both the SN and ST of aged animals when compared to young rats. Both structural and functional impairments were reversed in the aged animals after the supplementation of WA (p < 0.001). The present study clearly indicates that WA attenuates the ageing-mediated motor degenerative changes in the SN and ST of aged rats and ascertains its neuroprotective potential.

Redox modulating effects of grape juice during aging.

Background Polyphenols are known because of their phytochemical constituents having antioxidative properties. In this regard, grape juice is highly enriched with polyphenolic constituents, and its supplementation has been known to improve many health and age-associated diseases and risk factors. Our study was entirely dedicated to evaluating the positive effects of grape juice on young and old rats' erythrocytes and plasma. Methods Young (4 months) and old (24 months) male Wistar rats were given an oral dose of grape juice for 28 days. They were grouped into four categories (n = 6): Group I: young control rats; Group II: young grape juice treated rats; Group III: old control rats; Group IV: old treated rats. The treated groups were administered with 10 μL/g of grape juice according to body weight. The following biomarkers of antioxidant defense were measured: ferric reducing ability of plasma (FRAP), reactive oxygen species (ROS), plasma membrane redox system (PMRS), glutathione (GSH), osmotic fragility, and the decrease in lipid peroxidation measured in terms of malondialdehyde (MDA) levels. Results A significant increase (p<0.05) in antioxidant levels of FRAP, PMRS, and GSH and a significant decrease (p<0.05) in oxidized products such as ROS and MDA were seen in the treated rats in comparison to the controls. The decrease in ROS and rise in FRAP and PMRS levels suggest the ability of grape juice to combat oxidative stress effectively. Conclusion We propose the role of grape juice as a potent antioxidant because of its easy bioavailability and its role in combating stress. Our results also approve grape juice as a possible antiaging agent.