Contribution of senescent and reactive astrocytes on central nervous system inflammaging.

Astrocytes, the most predominant cells in the central nervous system (CNS), have well-recognized neuroprotective functions. However, during the CNS aging, astrocytes can become neurotoxic and contribute to chronic inflammation in age-associated brain deterioration and disease. Astrocytes are known to become senescent or reactive due to the exposure to stressful stimuli, in both cases they contribute to an impaired cognitive function through the production of pro-inflammatory mediators. Although both scenarios (senescence and reactive gliosis) have been studied independently, there are no direct studies comparing their secretomes simultaneously in the aging-brain. In this review we discuss the most recent studies in that respect, in order to analyze their simultaneous participation in brain aging.

Long-term sulforaphane-treatment restores redox homeostasis and prevents cognitive decline in middleaged female and male rats, but cannot revert previous damage in old animals.

Aging is a complex and detrimental process, which disrupts most organs and systems within the organisms. The nervous system is morphologically and functionally affected during normal aging, and oxidative stress has been involved in age-related damage, leading to cognitive decline and neurodegenerative processes. Sulforaphane (SFN) is a hormetin that activates the antioxidant and anti-inflammatory responses. So, we aimed to evaluate if SFN long-term treatment was able to prevent age-associated cognitive decline in adult and old female and male rats. Memory was evaluated in adult (15-month-old), and old (21-month-old) female and male Wistar rats after three months of SFN treatment. Young rats (4-month-old) were used as age controls. The antioxidant response induction, the redox state (GSH/GSSG), and oxidative damage were determined in the brain cortex (Cx) and hippocampus (Hc). Our results showed that SFN restored redox homeostasis in the Cx and Hc of adult rats, thus preventing cognitive decline in both sexes; however, the redox responses were not the same in males and females. Old rats were not able to recover their redox state as adults did, but they had a mild improvement. These results suggest that SFN mainly prevents rather than reverts neural damage; though, there might also be a range of opportunities to use hormetins like SFN, to improve redox modulation in old animals.

Association between physical activity and physical and functional performance in non-institutionalized Mexican older adults: a cohort study.

BACKGROUND: Aging is an independent risk factor for deterioration in functional capacity. Some studies have reported that physical activity (PA) improves functional capacity and physical performance among older adults (OA). Thus the objective of the present study was to assess the longitudinal association between PA and functional and physical performance in non-institutionalized OA. METHODS: A longitudinal analysis using data from the Frailty, Dynapenia and Sarcopenia in Mexican adults (FRADYSMEX, by its Spanish acronym) cohort study was conducted. PA was assessed through the Community Healthy Activities Model Program for Seniors (CHAMPS) instrument. Functionality was measured with the Barthel index and the Lawton and Brody scale, while physical performance was measured with the Short Physical Performance Battery (SPPB). To evaluate the association between the level of PA and physical and functional performance as a continuous variable, a linear regression of mixed effects was performed. To assess PA and dependence in basic activities of the daily life (BADL), instrumental activities of the daily life (IADL), and low physical performance (PP), generalized estimation equation models [to compute odds ratios (OR) and 95% confidence intervals (95%CI)] were computed. RESULTS: Older people who performed moderate to vigorous-intensity PA had a lower risk of dependence in IADL (OR = 0.17; 95%CI: 0.10, 0.80) and lower risk of low PP (OR = 0.18; 95%CI: 0.11, 0.58) compared to those in lower categories of PA. CONCLUSIONS: Older adults living in the community who perform PA of moderate to vigorous intensity have a lower risk of dependence in BADL and IADL and have a lower risk of low PP.

Effect of long-term moderate-exercise combined with metformin-treatment on antioxidant enzymes activity and expression in the gastrocnemius of old female Wistar rats.

Oxidative stress is known to be involved in the etiology of sarcopenia, a progressive loss of muscle mass and force related to elderly incapacity. A successful intervention to prevent this condition has been exercise-based therapy. Metformin (MTF), an anti-diabetic drug with pleiotropic effects, is known to retain redox homeostasis. However, the combined use of MTF with exercise has shown controversial experimental results. Our research group has shown that MTF-treatment does not limit the benefits provided by exercise, probably by inducing a hormetic response. Hence, our aim was to evaluate the effect of exercise in combination with MTF-treatment on the redox state of old female Wistar rats. Animals were divided into six groups; three groups preformed exercise on a treadmill for 5 days/week for 20 months and the other three were sedentary. Also, two groups of each, exercised and sedentary animals were treated with MTF for 6 or 12 months correspondingly, beside the untreated groups. Rats were euthanized at 24 months. Muscular functionality was analyzed as the relation between the lean mass free of bone with respect to the grip strength. Superoxide dismutase, catalase, and glutathione peroxidase content, enzymatic activity and redox state were determined in the gastrocnemius muscle. Our results showed that the exercised group treated with MTF for 12 months presented higher GSH/GSSG rate and high antioxidant scavenging power in contrast to the MTF-treatment for 6 months, where the beneficial effect was less noticeable.

The Effect of MPP+ on the Viability of Primary Cortical Astrocytes Isolated from Female and Male Wistar Rats of Different Ages.

Although age is known to be the main risk for developing chronic and neurodegenerative diseases, those illnesses have a different prevalence depending on the sex. It has been questioned whether genetic and hormonal differences are preserved in primary cultures from individuals of different genders. Therefore, here we studied the susceptibility of astrocytes, obtained from female and male Wistar rats of different ages (newborn, 9 and 24 months-old), to the well-known toxin MPP+ after 2 weeks in vitro, at different concentrations and exposure times. Our results showed that there are no variances due to gender, but that there are important differences associated to age in terms of the viability against this toxin.

Correction to: The Effect of MPP+ on the Viability of Primary Cortical Astrocytes Isolated from Female and Male Wistar Rats of Different Ages.

The original version of this article unfortunately contained a mistake in author names. The given name and family name was swapped erroneously for the three authors and published incorrectly as Alarcon-Aguilar Adriana, Luna-Lopez Armando and Königsberg Mina.The author names should read as Adriana Alarcón-Aguilar, Armando Luna-López and Mina Königsberg.The original article has been corrected.

The secretory phenotype of senescent astrocytes isolated from Wistar newborn rats changes with anti-inflammatory drugs, but does not have a short-term effect on neuronal mitochondrial potential.

In the central nervous system (CNS), senescent astrocytes have been associated with neurodegeneration. Senescent cells secrete a complex mixture of pro-inflammatory factors, which are collectively called Senescence Associated Secretory Phenotype (SASP). The SASP components can vary depending on the cell type, senescence inducer and time. The SASP has been mainly studied in fibroblasts and epithelial cells, but little is known in the context of the CNS. Here, the SASP profile in senescent astrocytes isolated from Wistar newborn rats induced to senescence by oxidative stress or by proteasome inhibition was analyzed. Senescent astrocytes secreted predominantly chemokines and IL-1α, but no IL-6. The effect of the anti-inflammatory drugs, sulforaphane (SFN) and dehydroepiandrosterone (DHEA), on the SASP profile was evaluated. Our results showed that SFN and DHEA decreased IL-1α secretion while increasing IL-10, thus modifying the SASP to a less anti-inflammatory profile. Primary neurons were subjected to the conditioned media obtained from drug-treated senescent astrocytes, and their mitochondrial membrane potential was evaluated.

Relationship of inflammatory profile of elderly patients serum and senescence-associated secretory phenotype with human breast cancer cells proliferation: Role of IL6/IL8 ratio.

Aging is considered a systemic, chronic and low-grade inflammatory state, called "inflammaging", which has been contemplated as a risk factor for cancer development and progression in the elderly population. Cellular senescence is a multifactorial phenomenon of growth arrest and distorted function, which has been recognized as a contributor to aging. Senescent cells have an altered secretion pattern called Senescent Associated Secretory Phenotype (SASP), that comprise a complex mix of factors including cytokines, growth factors, chemokines and matrix metalloproteinases among others. The SASP secreted by accumulated senescent cells during old age has been related to local inflammation that leads to cellular transformation and therefore may be supporting the inflammaging process. Here, we evaluated if the pro-inflammatory profile within the serum obtained from elderly patients (EPS) was able to induce cellular proliferation in the breast cancer transformed cell line (MCF-7), in a similar way to the proliferation stimulated by the SASP obtained from WI-38 primary cells prematurely induced to senescence by oxidative stress (SIPS). At the same time, the participation of IL-6/IL-8 ratio was determined. Our results showed that not all the EPS increased MCF-7 proliferation. However, there was an interesting relationship between IL-6 and IL-8 concentrations, when the IL-6 was higher than IL-8. Similar results were found with SASP from SIPS-WI-38 on the MCF-7 proliferation. Although it is known that those cytokines are fundamental factors to induce proliferation; the occurrence of other components in the cellular microenvironment is necessary to carry out this effect.

Bik subcellular localization in response to oxidative stress induced by chemotherapy, in Two different breast cancer cell lines and a Non-tumorigenic epithelial cell line.

Cancer chemotherapy remains one of the preferred therapeutic modalities against malignancies despite its damaging side effects. An expected outcome while utilizing chemotherapy is apoptosis induction. This is mainly regulated by a group of proteins known as the Bcl-2 family, usually found within the endoplasmic reticulum or the mitochondria. Recently, these proteins have been located in other sites and non-canonic functions have been unraveled. Bik is a pro-apoptotic protein, which becomes deregulated in cancer, and as apoptosis is associated with oxidative stress generation, our objective was to determine the subcellular localization of Bik either after a direct oxidative insult due to H2 O2 , or indirectly by cisplatin, an antineoplastic agent. Experiments were performed in two human transformed mammary gland cell lines MDA-MB-231 and MCF-7, and one non-tumorigenic epithelial cell line MCF-10A. Our results showed that in MCF-7, Bik is localized within the cytosol and that after oxidative stress treatment it translocates into the nucleus. However, in MDA-MB-231, Bik localizes in the nucleus and translocates to the cytosol. In MCF10A Bik did not change its cellular site after either treatment. Interestingly, MCF10A were more resistant to cisplatin than transformed cell lines. This is the first report showing that Bik is located in different cellular compartments depending on the cancer stage, and it has the ability to change its subcellular localization in response to oxidative stress. This is associated with increased sensitivity when exposed to toxic agents, thus rendering novel opportunities to study new therapeutic targets allowing the development of more active and less harmful agents.

Comparing the effects of two neurotoxins in cortical astrocytes obtained from rats of different ages: involvement of oxidative damage.

Oxidative stress has been recognized as a potential mediator of cell death. Astrocytes play an active role in brain physiology responding to harmful stimuli by activating astrogliosis, which in turn has been associated either with survival or degenerative events. The characterization of the mechanistic actions exerted by different toxins in astrocytes is essential to understand the brain function and pathology. As age plays a critical role in degenerative processes, the aim of this study was to determine whether the administration of equimolar concentrations of two neurotoxins evoking different toxic patterns can induce differential effects on primary astrocytes obtained either from newborn or adult rats, with particular emphasis on those events linked to oxidative stress as a potential source of damage. Primary cortical astrocyte cultures derived from rat brains were exposed to 1-methyl-4-phenylpyridinium (MPP+) or beta-amyloid peptide (ß-amyloid). Mitochondrial functionality and cell viability were determined as physiological parameters, whereas lipid and protein oxidation were used as markers of oxidative damage. The results of these experiments pointed towards a higher vulnerability to MPP + over ß-amyloid, on most of the tested markers. Hence, in order to allow a comprehensive evaluation of astrocytic responses against MPP + intoxication, a third astrocyte group was included for dose-response experiments: astrocytes derived from aged rats. The present data indicate that the differences associated with age were mainly found in astrocytes exposed to MPP + (25 and 50 µM) at 1-h treatment. Results are discussed in terms of the differential mechanisms involved in each model.

Cytokine profiling in senescent and reactive astrocytes: A systematic review.

Astrocytes play an important role in neuroinflammation by producing proinflammatory molecules. In response to various stressful stimuli, astrocytes can become senescent or reactive, both are present in age-associated cognitive impairment and other neurodegenerative diseases, and contribute to neuroinflammation. However, there are no studies that compare the cytokines secreted by these types of astrocytes in the brain during aging. Hence, we aimed to broaden the picture of the secretory profiles and to differentiate the variability between them. Therefore, a systematic review was conducted following the guidelines of the "Reporting Items for Systematic Review and Meta-Analyses". Only three studies that met the inclusion terms evaluated age-related cytokine secretion, however, no evaluation of senescence or gliosis was performed. Consequently, to increase the spectrum of the review, studies where those phenotypes were induced and cytokines determined were included. Although some cytokines were common for gliosis and senescence, some interesting differences were also found. The dissimilarities in cytokines secretion between these phenotypes could be studied in the future as potential markers.

Association between the risk of malnutrition and sarcopenia at 4.2 years of follow-up in community-dwelling older adults.

Introduction: Currently, there is only scarce evidence of a causal association between risk of malnutrition (RM) by the mini-nutritional assessment (MNA) and the incidence of sarcopenia. This study was designed to assess such an association at 4.2 years of follow-up in community-dwelling subjects over 60 years old. Methods: The data used were from the FraDySMex cohort study. The exposition variables were RM diagnosed by the long forma of the MNA (MNA-LF) and short form (MNA-SF). The last one included the body mass index and calf circumference at baseline, while sarcopenia was diagnosed by the EWGSOP2 at follow-up and taken as the response variable. Several covariates involved in the association were also considered. A multiple logistic regression analysis was performed to test the association. Results: At baseline, 27.0 and 37.9% of subjects had RM by the MNA-LF and MNA-SF, respectively. The incidence of sarcopenia was 13.7%. The fat mass variable significantly modified the association, so it was tested in each stratum. Two independent models showed that subjects with RM by the MNA-LF in the normal fat mass stratum were at a higher risk for developing sarcopenia at follow-up than those without RM (OR 9.28; IC 95% 1.57-54.76) after adjusting for age, sex, and waist circumference. No association was found for the excess fat mass stratum subjects. Subjects with RM by the MNA-SF in the excess fat mass stratum were more likely to develop sarcopenia at follow-up than those without RM by the MNA-SF (OR 3.67; IC 95% 1.29-10.43). This association was not found in the subjects in the normal fat mass stratum. Conclusion: The association was dependent on the variable fat mass. The two forms of the MNA should not be applied indistinctly with older adults. Based on these results, it is clear that the risk of malnutrition precedes the onset of sarcopenia.

[Hormesis: What doesn't kill you makes you stronger]. / Hormesis: lo que no mata, fortalece.

Living organisms have always had to cope with harsh environmental conditions and in order to survive, they have developed complex mechanisms to deal with them. These responses have been assembled in a concept called hormesis, which has been identified as an evolutionarily conserved process in which a low dose of a stressful stimulus activates an adaptive response that increases the resistance of the cell or organism to higher stress level. The main hormetic agents identified so far are irradiation, heat, heavy metals, antibiotics, ethanol, pro-oxidants, exercise and food restriction. The hormetic response involves the expression of genes that encode cytoprotective proteins such as chaperones like heat-shock proteins, antioxidant enzymes and growth factors. In this review we will discuss the hormetic response mainly during an oxidative challenge, and its relationship with senescence and aging, and some related diseases such as diabetes and neurodegeneration.

Development and validation of a Sarcopenia Geriatric Scale (SARCO-GS): a new short scale for the screening of sarcopenia.

Introduction: Sarcopenia is a highly prevalent disease associated with adverse outcomes such as falls, disability, and death. The current international consensuses agree that muscle strength, muscle mass, and gait speed must be included in the definition. However, these proposed criteria require objective measurements that are not available for most populations. Since the timely identification of sarcopenia is a priority, several subjective screening scales have been developed; however, they have some limitations due to their low sensitivity. The objective of this work was to develop and validate SARCO-GS, a new short scale to screen sarcopenia that is affordable, easy, and accessible for all clinical care settings. Methods and materials: The development of the SARCO-GS included four stages: (1) Review and analysis of documentary sources, (2) Contextualization of the theoretical model of sarcopenia, (3) Scale conformation, and (4) Reliability and validity analyses. SARCO-GS was validated in the FraDySMex study, which is a longitudinal cohort of community-dwelling adults. Results: In the studied population (n=852), the average age was 68.9 years (SD 10.21) and 80.1% of the participants were women. SARCO-GS is a seven-item scale with an innovative structure that included five subjective questions (gait speed, muscular strength, muscle mass) and two measurements of muscular strength and muscle mass (Chair stand test and calf circumference). The results regarding criterion validity showed that the cut-off point ≥ 3 had good sensitivity (77.68%) versus the EWGSOP2 consensus, with an adequate Area Under the Receiver Operating Characteristic (AUC) (0.73), in addition to showing higher values of sensitivity and AUC than SARC-F and SARC-CalF using as reference the same consensus. Furthermore, SARCO-GS presented good predictive validity for functional dependence (HR=2.22, p=0.046) and acceptable correlation with other related measurements (construct validity). Regarding reliability, the scale showed acceptable internal reliability (correlation between items and total score: 0.50 to 0.70). After the validation analysis, the scale was adapted to English. Conclusions: The SARCO-GS is a novel scale to screen sarcopenia with high sensitivity, good construct, predictive validity, and internal reliability that may be useful for health professionals in different clinical settings and for clinical research.

Incidence of the Risk of Malnutrition and Excess Fat Mass, and Gait Speed as Independent Associated Factors in Community-Dwelling Older Adults.

BACKGROUND AND AIMS: Only one cohort study exists on the incidence of the risk of malnutrition (RM) in older adults, though numerous cross-sectional reports, identified several risk factors associated with the prevalence and incidence of this condition. However, alterations in body composition and impaired physical performance as exposition variables of RM have not been explored. This study assessed the incidence of RM and determined its association with excess fat mass, low total lean tissue, gait speed, and handgrip strength as exposition variables for RM in community-dwelling older adults. METHODS: This is a secondary analysis of older adults (≥60 years) derived from the study "Frailty, dynapenia, and sarcopenia in Mexican adults (FraDySMex)", a prospective cohort project conducted from 2014 to 2019 in Mexico City. At baseline, volunteers underwent body composition analysis and physical performance tests. Several covariates were identified through comprehensive geriatric assessment. At baseline and follow-up, RM was assessed using the long form of the mini nutritional assessment (MNA-LF) scale. Associations between the exposition variables and RM were assessed by multiple logistic regression. RESULTS: The cohort included 241 subjects. The average age was 75.6 ± 7.8 years, and 83.4% were women. The mean follow-up period was 4.1 years, during which 28.6% of subjects developed RM. This condition was less likely to occur in those with an excess fat mass, even after adjusting for several covariates. Regarding total lean tissue, the unadjusted model showed that RM was more likely to occur in men and women with a low TLT by the TLTI classification, compared to the normal group. However, after adjusting for several covariates (models 1 and 2), the association lost significance. Results on the association between gait speed and RM showed that this condition was also more likely to occur in subjects with low gait speed, according to both the unadjusted and adjusted models. Similar results were found for RM in relation to low handgrip strength; however, after adjusting for the associated covariates, models 1 and 2 no longer reached the level of significance. CONCLUSIONS: RM diagnosed by MNA-LF was significantly less likely to occur among subjects with excess fat mass, and a significant association emerged between low gait speed and RM after 4.1 years of follow-up in these community-dwelling older adults. These results confirm the association between some alterations of body composition and impaired physical performance with the risk of malnutrition and highlight that excess fat mass and low gait speed precede the risk of malnutrition, not vice versa.

A low-intensity lifelong exercise routine changes miRNA expression in aging and prevents osteosarcopenic obesity by modulating inflammation.

Osteosarcopenic obesity (OSO) has been associated with increase immobility, falls, fractures, and other dysfunctions, which could increase mortality risk during aging. However, its etiology remains unknown. Recent studies revealed that sedentarism, fat gain, and epigenetic regulators are critical in its development. One effective intervention to prevent and treat OSO is exercise. Therefore, in the present study, by keeping rats in conditions of sedentarism and others under a low-intensity exercise routine, we established an experimental model of OSO. We determined the degree of sarcopenia, obesity, and osteopenia at different ages and analyzed the miRNA expression during the lifespan using miRNA microarrays from gastrocnemius muscle. Interestingly microarrays results showed that there is a set of miRNAs that changed their expression with exercise. The pathway enrichment analysis showed that these miRNAs are strongly associated with immune regulation. Further inflammatory profiles with IL-6/IL-10 and TNF-α/IL-10 ratios showed that exercised rats presented a lower pro-inflammatory profile than sedentary rats. Also, the body fat gain in the sedentary group increased the inflammatory profile, ultimately leading to muscle dysfunction. Exercise prevented strength loss over time and maintained skeletal muscle functionality over time. Differential expression of miRNAs suggests that they might participate in this process by regulating the inflammatory response associated with aging, thus preventing the development of OSO.

Chronic consumption of a hypercaloric diet increases neuroinflammation and brain senescence, promoting cognitive decline in middle-aged female Wistar rats.

Being overweight and obesity are world health problems, with a higher prevalence in women, defined as abnormal or excessive fat accumulation that increases the risk of chronic diseases. Excess energy leads to adipose expansion, generating hypertrophic adipocytes that produce various pro-inflammatory molecules. These molecules cause chronic low-intensity inflammation, affecting the organism's functioning and the central nervous system (CNS), inducing neuroinflammation. The neuroinflammatory response during obesity occurs in different structures of the CNS involved in memory and learning, such as the cortex and the hippocampus. Here we analyzed how obesity-related peripheral inflammation can affect CNS physiology, generating neuroinflammation and promoting cellular senescence establishment. Since some studies have shown an increase in senescent cells during aging, obesity, and neurodegenerative diseases, we proposed that cellular senescence participation may contribute to the cognitive decline in an obesity model of middle-aged female Wistar rats. The inflammatory state of 6 and 13 months-old female Wistar rats fed with a hypercaloric diet was measured in serum and CNS (cortex and hippocampus). Memory was evaluated using the novel object recognition (NOR) test; the presence of senescent markers was also determined. Our data suggest that the systemic inflammation generated by obesity induces a neuroinflammatory state in regions involved in learning and memory, with an increase in senescent markers, thus proposing senescence as a potential participant in the negative consequences of obesity in cognition.

Telomerase activity in response to mild oxidative stress.

We have analysed telomerase activity to determine whether it can be modified when BCL-2 is endogenously overexpressed in response to a mild oxidative stress treatment as part of a survival mechanism, in contrast with an exogenous bcl-2 overexpression due to a retroviral infection. Endogenous bcl-2 overexpression was induced after a low oxidative insult of H2O2 in mice primary lung fibroblasts and L929 cell, whereas bcl-2 exogenous overexpression was performed using a retroviral infection in L929 cells. Telomerase activity was quantified in Bcl-2 overexpressing cells by the TRAP assay. When the cells were treated with different H2O2 concentrations, only those exposed to 50 µM showed increased telomerase activity. This correlates with BCL-2 expression as part of the endogenous response to mild oxidative stress. Oxidative stress generated during the toxic mechanism of chemotherapeutic drugs might induce BCL-2 increment, enhancing telomerase activity and reactivating the oncogenic process. Clinical trials should take into consideration the possibility of telomerase activation following increased BCL-2 expression when treating patients with ROS (reactive oxygen species) generation by anti-cancer drugs.

Methotrexate Induces an Antioxidant Hormetic Response in Primary Rat Astrocytes.

Neurodegenerative diseases have increased worldwide in recent years. Their relationship with oxidative stress has motivated the research to find therapies and medications capable of suppressing oxidative damage and therefore slowing the progression of these diseases. Glutathione (GSH) is the most important cellular antioxidant in living beings and is responsible for regulating the cellular redox state. However, GSH cannot be administered by any route of administration, so molecules that increase its levels by activating Nrf2-ARE signaling pathway are explored; since Nrf2 regulates the main genes involved in GSH de novo synthesis and recycling. Astrocytes are the most important cell-type in the antioxidant cell response and are responsible for providing GSH and other substrates for neurons to have an efficient antioxidant response. Methotrexate (MTX) is an anti-inflammatory agent that has different cellular effects when administered at low or high concentrations. So in this study, we used MTX different concentrations and exposure times to induce a hormetic antioxidant response in rat primary astrocytes. Our results showed that 20 nM MTX pre-conditioning for 12 h augmented the GSH/GSSG ratio and protected cellular viability against a toxic MTX and H2O2 insult, which was abrogated when Nrf2 was inhibited by brusatol. Hence, MTX subsequent studies as a drug to counteract the progression of some stress-associated neurodegenerative diseases are suggested.

Moderate exercise combined with metformin-treatment improves mitochondrial bioenergetics of the quadriceps muscle of old female Wistar rats.

Sarcopenia is a syndrome that leads to physical disability and that deteriorates elderly people´s life quality. The etiology of sarcopenia is multifactorial, but mitochondrial dysfunction plays a paramount role in this pathology. Our research group has shown that the combined treatment of metformin (MTF) and exercise has beneficial effects for preventing muscle loss and fat accumulation, by modulating the redox state. To get an insight into the mechanism of the combined treatment, the mitochondrial bioenergetics was studied in the mitochondria isolated from old female Wistar rats quadriceps muscles. The animals were divided into six groups; three performed exercise on a treadmill for 5 days/week for 20 months, and the other three were sedentary. Also, two groups of each were treated with MTF for 6 or 12 months. The rats were euthanized at 24 months. The mitochondria were isolated and supercomplexes formation along with oxygen consumption, ATP synthesis, and ROS generation were evaluated. Our results showed that the combined treatment for 12 months increased the complex I and IV activities associated with the supercomplexes, simultaneously, ATP synthesis increased while ROS production decreased, indicating a tightly coupled mitochondria. The role of exercise plus the MTF treatment against sarcopenia in old muscles is discussed.