Ageing, Metabolic Dysfunction, and the Therapeutic Role of Antioxidants.

The gradual ageing of the world population has been accompanied by a dramatic increase in the prevalence of obesity and metabolic diseases, especially type 2 diabetes. The adipose tissue dysfunction associated with ageing and obesity shares many common physiological features, including increased oxidative stress and inflammation. Understanding the mechanisms responsible for adipose tissue dysfunction in obesity may help elucidate the processes that contribute to the metabolic disturbances that occur with ageing. This, in turn, may help identify therapeutic targets for the treatment of obesity and age-related metabolic disorders. Because oxidative stress plays a critical role in these pathological processes, antioxidant dietary interventions could be of therapeutic value for the prevention and/or treatment of age-related diseases and obesity and their complications. In this chapter, we review the molecular and cellular mechanisms by which obesity predisposes individuals to accelerated ageing. Additionally, we critically review the potential of antioxidant dietary interventions to counteract obesity and ageing.

Obesity and aging: Molecular mechanisms and therapeutic approaches.

The epidemic of obesity is a major challenge for health policymakers due to its far-reaching effects on population health and potentially overwhelming financial burden on healthcare systems. Obesity is associated with an increased risk of developing acute and chronic diseases, including hypertension, stroke, myocardial infarction, cardiovascular disease, diabetes, and cancer. Interestingly, the metabolic dysregulation associated with obesity is similar to that observed in normal aging, and substantial evidence suggests the potential of obesity to accelerate aging. Therefore, understanding the mechanism of fat tissue dysfunction in obesity could provide insights into the processes that contribute to the metabolic dysfunction associated with the aging process. Here, we review the molecular and cellular mechanisms underlying both obesity and aging, and how obesity and aging can predispose individuals to chronic health complications. The potential of lifestyle and pharmacological interventions to counter obesity and obesity-related pathologies, as well as aging, is also addressed.

The Good, the Bad, and the Ugly of ROS: New Insights on Aging and Aging-Related Diseases from Eukaryotic and Prokaryotic Model Organisms.

Aging is associated with the accumulation of cellular damage over the course of a lifetime. This process is promoted in large part by reactive oxygen species (ROS) generated via cellular metabolic and respiratory pathways. Pharmacological, nonpharmacological, and genetic interventions have been used to target cellular and mitochondrial networks in an effort to decipher aging and age-related disorders. While ROS historically have been viewed as a detrimental byproduct of normal metabolism and associated with several pathologies, recent research has revealed a more complex and beneficial role of ROS in regulating metabolism, development, and lifespan. In this review, we summarize the recent advances in ROS research, focusing on both the beneficial and harmful roles of ROS, many of which are conserved across species from bacteria to humans, in various aspects of cellular physiology. These studies provide a new context for our understanding of the parts ROS play in health and disease. Moreover, we highlight the utility of bacterial models to elucidate the molecular pathways by which ROS mediate aging and aging-related diseases.

Coordinated Pulses of mRNA and of Protein Translation or Degradation Produce EGF-Induced Protein Bursts.

Protein responses to extracellular cues are governed by gene transcription, mRNA degradation and translation, and protein degradation. In order to understand how these time-dependent processes cooperate to generate dynamic responses, we analyzed the response of human mammary cells to the epidermal growth factor (EGF). Integrating time-dependent transcript and protein data into a mathematical model, we inferred for several proteins their pre-and post-stimulus translation and degradation coefficients and found that they exhibit complex, time-dependent variation. Specifically, we identified strategies of protein production and degradation acting in concert to generate rapid, transient protein bursts in response to EGF. Remarkably, for some proteins, for which the response necessitates rapidly decreased abundance, cells exhibit a transient increase in the corresponding degradation coefficient. Our model and analysis allow inference of the kinetics of mRNA translation and protein degradation, without perturbing cells, and open a way to understanding the fundamental processes governing time-dependent protein abundance profiles.

TGF-ß-induced hCG-ß regulates redox homeostasis in glioma cells.

Transforming growth factor (TGF-ß) is associated with the progression of glioblastoma multiforme (GBM)-the most malignant of brain tumors. Since there is a structural homology between TGF-ß and human chorionic gonadotropin (hCG) and as both TGF-ß and hCG-ß are known regulators of oxidative stress and survival responses in a variety of tumors, the role of TGF-ß in the regulation of hCG-ß and its consequences on redox modulation of glioblastoma cells was investigated. A heightened hCG-ß level was observed in GBM tumors. TGF-ß treatment increased hCG-ß expression in glioma cell lines, and this heightened hCG-ß was found to regulate redox homeostasis in TGF-ß-treated glioma cells, as siRNA-mediated knockdown of hCG-ß (i) elevated reactive oxygen species (ROS) generation, (ii) decreased thioredoxin Trx1 expression and thioredoxin reductase (TrxR) activity, and (iii) abrogated expression of TP53-induced glycolysis and apoptosis regulator (TIGAR). Silencing of hCG-ß abrogated Smad2/3 levels, suggesting the existence of TGF-ß-hCG-ß cross-talk in glioma cells. siRNA-mediated inhibition of elevated TIGAR levels in TGF-ß-treated glioma cells was accompanied by an increase in ROS levels. As a farnesyltransferase inhibitor, Manumycin is known to induce glioma cell apoptosis in a ROS-dependent manner, and we investigated whether Manumycin could induce apoptosis in TGF-ß-treated cells with elevated hCG-ß exhibiting ROS-scavenging property. Manumycin-induced apoptosis in TGF-ß-treated cells was accompanied by elevated ROS levels and decreased expression of hCG-ß, Trx1, Smad2/3, and TIGAR. These findings indicate the existence of a previously unknown TGF-ß-hCG-ß link that regulates redox homeostasis in glioma cells.

IGF-1 induced HIF-1α-TLR9 cross talk regulates inflammatory responses in glioma.

The insulin-like growth factor (IGF-1) induces hypoxia inducible factor (HIF-1α) regulated genes in glioblastoma multiforme (GBM). As HIF-1α links inflammatory and oncogenic pathways in GBM, we investigated whether IGF-1 affects HIF-1α to regulate inflammatory response in glioma cells under normoxia. IGF-1 induced Ras and Calmodulin-dependent kinase II (CaMKII) regulated HIF-1α transcriptional activity in glioma cells. Increase in HIF-1α was concurrent with decreased Toll-like receptor (TLR9) and CXCR4 expression and elevated suppressor of cytokine signaling (SOCS3) levels. Interestingly, while synthetic CpG containing oligodeoxynucleotide TLR9 agonist (CpG DNA) decreased IGF-1 mediated increase in HIF-1α activity, siRNA mediated knockdown of HIF-1α decreased TLR9 levels. This suggested that IGF-1 induced HIF-1α-TLR9 axis is regulated by both positive and negative feedback loops. Importantly, TLR9 agonist reversed the effect of IGF-1 on CXCR4 and SOCS3 expression. While knockdown of HIF-1α abrogated IGF-1 mediated increase in SOCS3 it elevated IGF-1 induced decrease in CXCR4 levels. Thus HIF-1α positively and negatively regulates SOCS3 and CXCR4 expression respectively, in glioma cells. Though TLR9 agonist had no additive effect on IGF-1 mediated increase in pro-inflammatory cytokines IL-1ß, IL-6 and IL-8, treatment with TLR9 agonist alone elevated expression of these pro-inflammatory cytokines. Our studies indicate that a complex HIF-1α-TLR9 cross-talk sustains a self-regulating cycle of inflammatory response through intrinsic negative and positive feedback mechanisms.

Protein nitration, lipid peroxidation and DNA damage at high altitude in acclimatized lowlanders and native highlanders: relation with oxygen consumption.

Reactive oxygen and nitrogen species have been reported to be increased due to hypobaric hypoxia. It was hypothesized that lowlanders are more susceptible to protein nitration, lipid peroxidation and DNA damage at high altitude than highlanders and formation of these biomarkers may have strong correlation with oxygen consumption. Male volunteers were randomly selected and categorized into 3 groups, i.e. lowlanders at sea level (LL-SL, n=10), lowlanders at an altitude of 4560 m (LL-HA, n=10) and highlanders (HAN, n=10). Volunteers performed maximal aerobic exercise. Resting and post-exercise blood samples were taken at sea level and high altitude. Both resting and maximum oxygen consumption showed positive correlation with stress markers. LL-HA showed increased 3-nitrotyrosine and lipid hydroperoxide than LL-SL at rest. 3-Nitrotyrosine and lipid hydroperoxide increased after exercise in 3 groups, but percentage increase was higher in HAN than LL-SL and LL-HA. LL-SL and HAN showed significant DNA damage after exercise. Results indicate that resting oxygen consumption is positively correlated with nitrosative and oxidative stress markers irrespective of environmental condition and adaptation levels. Lowlanders have shown higher susceptibility to hypoxic insult than highlanders at rest, but when subjected to exercise test, they showed better tolerance to hypoxia than highlanders.

Antioxidant and oxidative stress responses of sojourners at high altitude in different climatic temperatures.

High altitude (HA) is a multi-stressor environment comprising hypobaric hypoxia and cold. Climatic temperature varies with seasonal variation at HA. The present study was undertaken to investigate the effect of ambient temperature on antioxidant profile among sojourners at HA. The study was conducted on sojourners exposed to an altitude of 4,560 m in two different seasons and categorized into two groups (SOJ 1, n=63, ambient temp. at HA: -6 degree to +10degreeC; SOJ 2, n=81, ambient temp. at HA: 3degree-22degreeC). Blood was collected at sea level (SL) and after 4 weeks of HA exposure. Antioxidant enzymes showed significant upregulation in SOJ 2 at HA. In SOJ 1, superoxide dismutase and glutathione peroxidase showed significant upregulation but catalase and glutathione reductase showed significant decrease at HA. Non-enzymatic antioxidants showed significant reduction in SOJ 1 whereas a sustained antioxidant profile was observed in SOJ 2 at HA. Oxidative stress markers showed higher levels in SOJ 1 than SOJ 2 at HA. Differences observed between SOJ 1 and SOJ 2 at HA may be the consequence of different environmental temperatures. Cold stress was higher in SOJ 1 as evidenced from the significantly lower oral temperature in SOJ 1 as compared to SOJ 2. Cold- and hypoxia-induced increase in energy expenditure was significantly high in SOJ 1 than SOJ 2. To conclude, chronic exposure to hypoxia in moderate climatic temperature has a potential preconditioning effect on antioxidant system, but exposure to both cold and hypoxia causes greater oxidative stress due to altered metabolic rate.

Total antioxidant status at high altitude in lowlanders and native highlanders: role of uric acid.

Hypobaric hypoxia causes oxidative stress and the antioxidant system of the body plays a vital role in controlling it. Urate contributes up to two-thirds of the antioxidant capacity of human blood. The urate production is catalyzed by xanthine oxidase with a concomitant release of free radicals. This study was designed to appraise the role of urate as an antioxidant at high altitude. The study was conducted on 92 male lowlanders and 66 highlanders after ascent to high altitude at 4560 m. Blood was collected at sea level and after 4 weeks of high altitude exposure. In lowlanders, a significant increase in levels of hydroperoxide (551.4 +/- 4.2 micromol/mL vs. 582.0 +/- 3.55, p < 0.001], protein carbonyl (2.4 +/- 0.11 micromol/mL/mg protein vs. 3.03 +/- 0.11, p < 0.001), TAS (1.02 +/- 0.01 mmol/L vs. 1.19 +/- 0.02, p < 0.001), and UA (298.0 +/- 6.68 micromol/L vs. 383.0 +/- 6.55, p < 0.001) was observed at high altitude. These measurements were significantly lower in highlanders than in lowlanders at high altitude. Total antioxidant status (TAS) and uric acid (UA) showed a positive correlation in lowlanders at sea level and in highlanders at high altitude. Hydroperoxide and TAS also showed a positive correlation in both groups at high altitude. This indicates increased oxidative stress at high altitude despite an increase in antioxidant capacity in lowlanders. To conclude, a hypoxia-induced increase in UA contributes an appreciable portion of plasma total antioxidant capacity, but may not be effective in preventing oxidative stress at high altitude.

Different adaptation patterns of antioxidant system in natives and sojourners at high altitude.

Comparative studies on the adaptation pattern of antioxidant status among high altitude natives and acclimatized sojourners are very scanty. The aim of the present study was to compare the differences in antioxidant profile between two groups of active male volunteers, i.e. native highlanders (HAN, n=66) in their natural hypoxic environment with that of sojourners (SOJ, n=81) from sea level (SL) after 4 weeks of stay at an altitude of 4560m. Blood samples of SOJ were collected at SL and HA. Same was collected from HAN once at HA. HAN had significantly higher SOD activity and significantly lower catalase, GPX and GR activities than SOJ at HA. Ratio of GSH/GSSG was also significantly higher in HAN than SOJ at HA. In SOJ, antioxidant profile showed an upregulation after HA stay but it was not effective to reduce the levels of oxidative stress markers. Therefore, it can be stated that lifelong exposure to hypoxia has beneficial adaptive effects on antioxidant system in HAN. Similarly, acclimatization to HA also has beneficial preconditioning effects on antioxidant system in SOJ, but, may not be sufficient to ameliorate oxidative stress completely. Transient increase in metabolic rate due to hypoxia may be a causative factor for excess free radical generation among sojourners at HA.

Antioxidant and redox status after maximal aerobic exercise at high altitude in acclimatized lowlanders and native highlanders.

Exercise-induced increase in oxygen consumption leads to oxidative stress. On the contrary, hypoxia triggers oxidative stress despite decreased oxygen flux. Therefore, exercise under hypoxia may aggravate oxidative damage. Highlanders are expected to have better antioxidant capacity than lowlanders as a result of adaptation to hypoxia. The present study was undertaken to investigate the effect of exercise on antioxidant system in lowlanders and highlanders at high altitudes (HA). This study was conducted on active male volunteers, randomly selected and categorized into three groups, i.e., lowlanders tested at sea level (LL-SL, n = 35), lowlanders tested at altitude of 4560 m (LL-HA, n = 35) and native highlanders tested (HAN, n = 20) at the same height. Volunteers performed maximal exercise until exhaustion. Blood samples were collected before and after exercise. Both LL-SL and HAN had shown similar VO2max, which was significantly higher than LL-HA. GSH/GSSG ratio significantly increased in LL-SL and decreased in HAN after exercise. With exercise there were a decrease in superoxide dismutase and increase in glutathione peroxidase and catalase activities in HAN. Therefore, the results have suggested that HAN are more susceptible to oxidative stress when subjected to high-intensity exercise than lowlanders. The cumulative effect of higher VO2max and longer duration of exercise in hypoxia may be the reason of higher level of oxidative insult among HAN. Comparatively better management of antioxidant system observed in lowlanders at HA may be explained by the lower VO2max and shorter duration of exercise in hypoxia.

Improvement of glutathione and total antioxidant status with yoga.

OBJECTIVE: Several studies suggest that yoga can decrease oxidative stress. However reports are scanty regarding whether yoga training can improve the glutathione level of individual. This study is designed to appraise the role of yoga in maintaining glutathione (reduced and oxidized) levels and antioxidant status. STUDY DESIGN: This study was conducted on healthy male volunteers from the Indian Navy, who were divided into two groups--a yoga (n = 30) group and a control (n = 21) group. The yoga group was trained in yoga for 6 months. The yoga schedule consisted of prayers, asana, pranayama, and meditation. The control group practiced routine physical training exercise for 6 months. Blood samples were collected when the volunteers were in fasting condition before and after completion of 6-month training period. Reduced and oxidized glutathione, glutathione reductase activity and total antioxidant status (TAS) were estimated. RESULTS: Reduced glutathione level increased significantly (p < 0.05) in the yoga group after completion of training. Glutathione reductase activity increased significantly in the control group (p < 0.05). TAS increased significantly (p < 0.001) in the yoga group and decreased significantly (p < 0.001) in the control group. CONCLUSIONS: Regular practice of yoga can maintain or improve antioxidant level of the body. The clinical relevance is that yoga practice can be used to maintain the antioxidant defense system under stressful conditions of training as observed in the case of soldiers and athletes.

Evaluation of possible goitrogenic and anti-thyroidal effect of nitrate, a potential environmental pollutant.

Nitrate is a wide spread contaminant of ground and surface water. The source of nitrate in the ground water may be from run off or seepage from fertilized soil, municipal or industrial waste water, land fills, septic system, urban drainage or decaying plants. Human and animal systems are affected severely on nitrate exposure. The study was to investigate the effect of dietary nitrate exposure on the thyroid status along with the state of iodine nutrition. Rats were fed diet containing 3% potassium nitrate (KNO3) for 4 weeks and then thyroid status was evaluated by thyroid gland weight, urinary iodine excretion pattern, thyroid peroxidase (TPO) activity, serum levels of total thyroxine (T4), triiodothyronine (T3) and thyroid stimulating hormone (TSH) concentrations. In nitrate treated animals, the weight of thyroid gland was increased significantly (P<0.001) while thyroid peroxidase activity (P<0.01), serum T4 (P<0.01) and serum T3 levels (P<0.001) were reduced; but serum TSH level was increased (P<0.001) along with slightly elevated iodine excretion level (P<0.001) in comparison to control animals. The overall results indicated the development of a relative state of functional hypothyroidism with enlarged thyroid after nitrate exposure. This study can explain a part for the persistence of residual goitre in the post-salt iodization phase.