Changes in Leptin Gene Expression and Receptor in Aged Male Rats Following Continuous Training with and without Calorie Restriction

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Methods : this experimental study was conducted with a posttest design and a control group. The subjects consisted of 24 aged male rats, purposefully assigned to four groups based on weight: control, calorie restriction, continuous exercise, and continuous exercise with calorie restriction. The training duration was six weeks, comprising five 50-minute sessions per week, totaling 30 sessions. Each session included three phases: warm-up, cool-down (each featuring 6–8 m/min), and the main exercise (continuous running at 10 m/min on a specialized treadmill). The calorie restriction program involved reducing food intake by 30%. 48 hours after the last training session and an 8-hour fast, the rats were anesthetized with a combination of ketamine (90 mg/kg) and xylazine (10 mg/kg). Results : continuous training combined with calorie restriction significantly affected leptin gene expression and its receptor in aged male rats (P0.05). Conclusion : These findings demonstrate that continuous exercise and calorie restriction can influence leptin gene expression and its receptor. Leptin Continuous Training Calorie Restriction atherosclerosis Figures Figure 1 Introduction Cardiovascular disease (CVD) is currently the leading cause of death worldwide. CVD-induced deaths increased from 17.8 to 18.6 million between 2017 and 2019, reflecting a significant rise in mortality associated with this condition. This alarming increase highlights the growing impact of CVD on global health [1, 2]. Atherosclerosis, or coronary artery disease, is one of the most prominent forms of CVD. It is a complex clinical condition characterized by an inflammatory response to vascular injury. The injury causes endothelial dysfunction in the early stages of atherosclerosis. Over time, the accumulation and transformation of lipids, inflammatory cells, smooth muscle cells, and necrotic cell debris in the intima layer (beneath the single layer of endothelial cells) lead to the formation of fatty streaks, plaque buildup, increased vulnerability of blood vessels, and eventually, vascular rupture [3, 4]. The primary component of this disease is the accumulation of fat and inflammation in the large arteries, which can reduce blood flow in the lumen by more than 50%. Ultimately, it may lead to clinical complications such as myocardial infarction, ischemic cardiomyopathy (the most common cause of heart failure), and stroke [5, 6]. Atherosclerosis, predominantly a disease of older adults, is exacerbated by increased life expectancy and a growing aging population [7]. Factors such as cellular senescence [8], weight gain (particularly the accumulation of visceral fat) in old age [9], and insufficient physical activity—falling short of the levels recommended by the World Health Organization for many elderly individuals—raise significant concerns regarding the future burden of CVD [10]. Consequently, there is a pressing need to focus on the pathology of atherosclerosis and explore effective prevention strategies. In general, advancing age, obesity, and a sedentary lifestyle are the most critical factors contributing to atherosclerosis [11]. Reportedly, a 10 kg increase in body weight raises the risk of coronary artery disease by 12% while also increasing systolic blood pressure by approximately 3 mm Hg and diastolic blood pressure by 2.3 mm Hg [12]. The link between obesity and cardiovascular risk is primarily driven by the increased body fat percentage that accompanies weight gain. Adipose tissue, a highly active and metabolically significant endocrine organ, secretes biologically active molecules known as adipokines or adipocytokines. Among these, important adipokines that are elevated in visceral obesity include leptin, resistin, retinol-binding protein-4, angiopoietin-like protein 2 (AngptL-2), interleukin-6, and Monocyte Chemotactic Protein-1 (MCP-1). These adipokines contribute to systemic inflammation, endothelial dysfunction, hypercoagulability, and insulin resistance, all of which are key contributors to atherosclerosis [13]. Among these adipokines, however, leptin's specific role in atherosclerosis development remains ambiguous. Leptin, a 16 kDa peptide hormone produced by the LEP gene, is primarily secreted from adipose tissue. Leptin binding to its specific receptor (LEPR) in the arcuate nucleus of the hypothalamus regulates food intake, energy expenditure, and hormone secretion. Therefore, leptin plays a crucial role in regulating body weight, energy balance, inflammatory responses, immune function, angiogenesis, vascular function, bone homeostasis, and reproduction [14]. Circulating leptin levels (typically around 16 ng/ml) reflect the amount of energy stored in adipose tissue and are directly related to the degree of obesity. Increased adipose tissue may lead to leptin resistance and elevated leptin levels [15]. Various factors, including inflammatory cytokines, glucocorticoids, and insulin, regulate the expression and secretion of leptin. Conversely, sympathetic norepinephrine release and β-adrenergic receptor activation in adipose tissue are key factors in reducing leptin gene expression in response to leptin injection [16]. Notably, leptin is too large to cross the blood-brain barrier passively; hence, it is transported through a saturable and regulated transport system. While leptin reaches most central LepRb neurons via this system or through the cerebrospinal fluid, arcuate nucleus LepRb neurons can uniquely detect changes in circulating leptin independent of the blood-brain barrier, responding to these changes with time- and dose-dependent sensitivity [16]. Previous studies have indicated that patients with CVD exhibit elevated serum leptin levels compared to healthy individuals [17]. These findings indicate that leptin may facilitate an inflammatory state that enhances monocyte recruitment to the arterial intima and stimulates pro-inflammatory cytokine production, thereby positioning leptin as a potential mediator of atherosclerosis [18]. Additionally, leptin can induce a prothrombotic state by increasing the expression of Plasminogen Activator Inhibitor-1 (PAI-1) a major endogenous inhibitor of fibrinolysis [19]. More importantly, age-related loss of leptin responsiveness in stress signaling and eNOS phosphorylation may cause changes in cardiac geometry and function, particularly in obese and older people [20]. There are six leptin receptor (LepR) isoforms, all of which are alternative splicing products of the db gene transcript. These receptors exhibit different expression patterns across various tissues. Although they share identical extracellular and membrane-spanning domains, their intracellular domains differ in length. Type 2 leptin receptors in peripheral tissues play a significant role in regulating energy and body weight in these tissues upon activation [21]. Lifestyle interventions, such as low-calorie diets and regular exercise, are commonly recommended as primary strategies to improve the health of individuals at risk for atherosclerosis [22]. The combination of dietary and exercise interventions is expected to have additive effects. Meta-analyses in this field suggest that combining exercise training with calorie restriction may be more effective than either intervention alone [23, 24]. However, these results are not always consistent. For instance, a recently published meta-analysis did not find significant reductions in inflammatory marker concentrations following concurrent diet and exercise interventions compared to diet alone [25]. The conflicting results of these studies highlight the need for further research with more controlled variables. Therefore, this research aimed to investigate and compare the changes in serum levels of leptin, leptin receptor, and PAI-1 following a period of continuous training with and without calorie restriction in aged male rats. Methodology Experimental Animals This experiment was approved by the Laboratory Animal Ethics Committee of Bojnord University. The study involved 24 male Wistar rats with a mean age of 26 months and an average weight of 200–250 grams. The rats were purchased from the Institute Pasteur of Tehran. In order to ensure proper acclimatization, all rats were exposed to a 12-hour light/12-hour dark cycle, maintained at an ambient temperature of 23–25°C and an ambient humidity of 55–65%. Experimental Design and Diet The statistical sample was randomly assigned to four groups, each consisting of six rats: continuous training, continuous training with calorie restriction, calorie restriction, and control. The maximum running capacity (MRC) test was performed to assess the aerobic performance of the rats. The test was conducted until the point of fatigue, i.e., after two stimulations with a soft shoulder and no response from the rat. The general outline of this test is depicted in Figure 1. This test was conducted at baseline and the end of the study. The average speed in the continuous training group and continuous training with calorie restriction group was 35.66 m/min and 37.33 m/min, respectively. Moreover, the average test duration was 21.16 minutes and 17.26 minutes, respectively. During the study, each rat underwent a six-week training program based on its group assignment, consisting of five 50-minute sessions per week. During the first two weeks, the rats were familiarized with the rodent-specific treadmill by exercising for 5 to 10 minutes at a speed of 5 to 10 meters per minute. Each training session included a 5-minute warm-up and cool-down period at an intensity of 30% of the maximum speed (10 meters per minute). The continuous training started with 10 minutes per session at an intensity of 60% of the maximum speed (20 meters per minute) during the first week. It gradually increased to 40 minutes per session by the sixth week while maintaining the same intensity. Maximum speed during maximum oxygen consumption was determined using the test protocol by Leonardo et al. (2007) to establish training intensity. This protocol involved a 3-minute warm-up at a speed of 5 meters per minute, after which the treadmill speed was increased by 4 meters per minute every two minutes. The maximum speed was defined as the point where laboratory rats could no longer maintain a steady pace for at least 30 seconds and failed to continue running despite an increase in speed [26] . The required pelleted feed for the rats was purchased from the knowledge-based company Med Zist Samaneh Pishro. The amount of food provided to the rats, excluding the calorie-restricted groups, was 10 grams of pellets per 100 grams of body weight, along with 10 to 15 milliliters of water. However, the calorie-restricted groups received 30% less food than the ad libitum amount. The percentage of food reduction was adjusted based on subsequent weight measurements, ultimately reaching a 20% reduction in food intake. Rat Dissection and Tissue Sampling Forty-eight hours after the final training session and an 8-hour fast, the animals were anesthetized using a combination of ketamine (90 mg/kg) and xylazine (10 mg/kg). The abdominal cavity was subsequently opened, and upon locating the aorta, a suitably sized section was excised. Additionally, a portion of visceral fat was collected, immediately placed in a labeled 1.8 mL microtube, and rapidly frozen in liquid nitrogen before it was stored at -80°C. cDNA Synthesis for PCR (Reverse-Transcriptase) Reaction The reverse-transcriptase PCR (RT-PCR) is one of the most sensitive methods for detecting RNA. The kit used for cDNA synthesis provided a complete system for synthesizing the first complementary strand of cDNA from RNA, capable of synthesizing cDNA up to 13 kilobases in length. The kit included both oligo(dT) and random hexamer primers. In order to verify the accuracy of cDNA synthesis, a PCR reaction was conducted using primers specific to the GPI control gene. The PCR product was subsequently visualized on an agarose gel, where clear bands were observed at approximately 350 base pairs for the GPI gene, with no secondary bands present, confirming the quality of the synthesized cDNA. After cDNA synthesis, the quality of each extracted sample was measured using a nanodrop spectrophotometer at a wavelength of 260 nm. All samples had concentrations exceeding 450 ng/μL. Primer Design for RQ-PCR (Relative Quantitation) Reaction Once the nucleotide sequence for the target gene was acquired, BLAST software was used on the NCBI website, as per the research by Baldus et al., to ascertain primer specificity and evaluate the homology of the primer sequence with registered genetic sequences. Table 1. Sequence of primers Gene name primers Sequence PAI-1 Forward TGAGAGAGGGCAAAGTGGTT Reverse ACGTGTCCACTGCTCACTTA Leptin Forward GAC ATT TCA CAC ACG CAG TC Reverse GAG GAG GTC TCG CAG GTT Leptin receptor Forward CAC TGT TAA TTT CAC ACC AGA G Reverse GTC ATT CAA ACC ATA GTT TAG G Real-Time PCR Analysis Real-time PCR was conducted using an ABI thermocycler. The PCR reactions were performed after the primers had been prepared. The ΔCT values for the Aplin, APJ, and ACE2 genes were calculated for both the control (healthy) and experimental samples. Expression levels were assessed by calculating the relative quantity (Fold Change) using the appropriate formula. Data were analyzed using one-way analysis of variance (ANOVA) with SPSS software version 26. Statistical Results Initially, the height and weight data of the rats were analyzed using descriptive statistics, including mean, standard deviation, minimum, and maximum (Table 2). The normality of the research variables was assessed using the Shapiro-Wilk and Lune tests. The results indicated that leptin levels and leptin receptor expression did not follow a normal distribution, while PAI-1 levels were normally distributed. Consequently, the Kruskal-Wallis test, a non-parametric method, was employed to analyze leptin and leptin receptor data, whereas one-way ANOVA as a parametric test was used for the PAI-1 data. Table 3 presents the changes in data related to leptin and its receptor. Table 3. Kruskal-Wallis test results for leptin and leptin receptor Variables Groups Mean SD Statistic Df Sig Leptin Continuous Exercise 12.35 4.93 18.274 3 *0.001 Continuous Exercise with Calorie Restriction 2.59 1.33 Calorie Restriction 7.29 6.06 Control 0.99 0.005 Leptin receptor Continuous Exercise 0.156 0.07 12.96 3 *0.005 Continuous Exercise with Calorie Restriction 0.77 0.54 Calorie Restriction 1.32 0.97 Control 0.99 0.005 The results of the Kruskal-Wallis test revealed that the significance level for both leptin levels and leptin receptor expression was less than 0.05, indicating significant changes in these variables. Moreover, Bonferroni's post hoc test was employed to identify the specific locations of these differences. The findings from this post hoc analysis are presented in Table 4. Table 4. Bonferroni post hoc test results Variables Groups 1 Groups 2 Df Sig Leptin Continuous Exercise with Calorie Restriction Continuous Exercise 9.76 *0.001 Calorie Restriction 5.05 *0.039 Control 11.35 *0.001 Continuous Exercise Calorie Restriction 4.70 0.053 Control 1.59 0.494 Calorie Restriction Control 6.30 *0.012 Leptin receptor Continuous Exercise with Calorie Restriction Continuous Exercise 6.17 0.069 Calorie Restriction 1.17 *0.002 Control 0.84 0.017 Continuous Exercise Calorie Restriction 0.55 0.100 Control 0.22 0.496 Calorie Restriction Control 0.33 0.315 Additionally, in order to investigate the changes in PAI-1 expression in the aorta tissue of rats, a one-way analysis of variance test was used, as displayed in Table 5. Table 5. One-way ANOVA results of PAI-1 expression Source of changes Sum of squares Df Mean square F Sig Between-group 0.908 3 0.303 1.46 0.254 Within-group 4.126 20 0.206 Total 5.034 23 The results of one-way ANOVA indicated that the significance level for PAI-1 levels was greater than 0.05, suggesting the absence of any significant changes in this variable. Discussion and Conclusion Over the past decade, numerous studies have explored the diverse roles of leptin and its potential effects on the development or exacerbation of conditions such as obesity, diabetes, metabolic syndrome, obstructive pulmonary disease, bone density issues, and CVD. These studies aim to mitigate the spread of these diseases and improve lifestyle quality. The present study aligns with previous research, seeking to determine whether combining exercise and calorie restriction is more effective than implementing either intervention alone. In summary, the results of this study demonstrate a significant difference in the effects of continuous exercise combined with calorie restriction versus either intervention alone on leptin gene expression in aged male rats. Specifically, both calorie restriction and continuous exercise with calorie restriction significantly increased leptin gene expression in these rats. Additionally, a significant difference was observed between the effects of continuous training with and without calorie restriction on leptin receptor gene expression. Indeed, continuous training combined with calorie restriction significantly reduced leptin receptor gene expression. Finally, no significant difference was found between the effects of continuous exercise with and without calorie restriction on PAI-1 expression in the aortic tissue of aged male rats. These findings contrast with previous reports, such as the study by Tremblay et al. (2019), which described decreased leptin levels following combined interventions [27]. A critical factor to consider is that, in obese populations, the high volume of adipose tissue and subsequent elevated secretion of adipokines often result in leptin resistance. Under these conditions, the hypothalamus becomes less responsive to leptin, leading to persistent hunger and unchanged food intake despite the abundant energy stored as fat [28]. In such cases, regular exercise can reduce adipose tissue and, consequently, decrease adipokine secretion, leading to lower leptin levels. It has been suggested that the adipokine response to exercise is correlated with the amount of adipose tissue [29]. However, in populations with normal adipose tissue levels, the significant increase in leptin levels observed after combined interventions might be attributed to the metabolic demands imposed by reduced caloric intake and increased energy expenditure due to physical activity. Under conditions that elevate the metabolic rate, such as exercise, leptin secretion by adipocytes gradually increases. Leptin signaling through its receptors in the skeletal muscle activates the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α). In physiological conditions, increased leptin concentration leads to higher expression of leptin receptors and enhanced leptin reabsorption, creating a positive feedback loop that strengthens the leptin signal [30]. The enhanced effects of leptin during continuous exercise coupled with calorie restriction are noteworthy. Previous studies have indicated that weight loss efforts, regardless of whether they involve obese individuals or those without a history of obesity, frequently result in a disproportionate decrease in energy expenditure compared to the reduction in body mass. In such cases, the tone of the sympathetic nervous system and the circulating concentrations of leptin and bioactive thyroid hormones decrease. In contrast, the tone of the parasympathetic nervous system increases, ultimately resulting in unsuccessful weight loss attempts. Decreased leptin levels signal the body to restore energy balance by increasing appetite and decreasing energy expenditure [31]. Conversely, elevated leptin levels suppress appetite by reducing hypothalamic neuropeptide Y (an appetite stimulator) and increasing α-MSH (an appetite suppressant), as well as by enhancing energy expenditure through increased secretion of TRH and GnRH [32]. In this context, it has been shown that lean mice receiving chronic leptin injections lose fat mass in a dose-dependent manner (within the physiological range). Leptin treatment has also been reported to reverse the changes in circulating thyroid hormone and corticosterone levels that are typically associated with food deprivation [33]. This natural increase in leptin levels can have beneficial effects on atherosclerosis. Leptin treatment appears to reduce vascular inflammation and endothelial-to-mesenchymal transition, a key factor in atherosclerosis development. Previous research has demonstrated that leptin treatment of endothelial cells significantly increases the expression of pro-inflammatory markers, such as intercellular adhesion molecule-1 and E-selectin, at the mRNA level. This may suggest that leptin mediates anti-inflammatory properties in the endothelium [34]. Moreover, plasma leptin levels appear to be inversely correlated with ObR expression in the arcuate nucleus of the hypothalamus, consistent with the changes in leptin receptor levels observed in the present study [35]. While previous studies suggest that increased leptin levels could be a direct outcome of exercise, the results of this research clearly show that combined interventions can have synergistic effects [36]. It has been proposed that when leptin binds to its soluble receptor, it is unable to activate membrane receptors, leading to the hypothesis that the soluble form of leptin receptors may be involved in leptin resistance [36]. Additionally, the results indicated no significant difference between the effects of continuous training with and without calorie restriction on PAI-1 expression in the aortic tissue of aged male rats. Within-group analysis showed increased, though insignificant, PAI-1 gene expression following exercise and calorie restriction. These findings align with previous studies reporting improvements in PAI-1 fibrinolytic activity following aerobic exercise [37]. However, the absence of a significant difference in the current study may be attributed to variations in baseline PAI-1 levels, which impedes drawing definitive conclusions. The small sample size may further account for this outcome; therefore, additional research is recommended to explore this issue in more depth. Declarations Compliance with Ethical Standards The authors declare there no conflict of interest. All procedures involving animals were carried out in compliance with the relevant national regulations, including the Iranian Animal Welfare Law, and the ethical guidelines set by the Bioethics Committee of Birjand University, under approval number [IR.UB.REC.1401.010]. Funding This research received no external funding. Authors' contributions All authors contributed equally to the preparation of this article. Acknowledgments This research project was conducted in the Animal Laboratory of the Faculty of Physical Education at Ferdowsi University of Mashhad. 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Yasari, S., et al., Exercise training decreases plasma leptin levels and the expression of hepatic leptin receptor-a,-b, and,-e in rats. Molecular and Cellular Biochemistry, 2009. 324: p. 13-20. Habibian, M. and S.A. Rezaye Mousavi, The Effects of Regular Aerobic Exercise on the Serum levels of Plasminogen Activator Inhibitor-1 and Adiponectin in Obese Girls. Journal of Inflammatory Diseases, 2019. 23(5): p. 418-429. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6493746","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":580439611,"identity":"e319f344-41f8-4466-81a2-56c38fb935e2","order_by":0,"name":"Fatemeh Zafarani","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Fatemeh","middleName":"","lastName":"Zafarani","suffix":""},{"id":580439612,"identity":"af30fc1d-beff-4189-816f-6c2aae9ba9dc","order_by":1,"name":"Mahtab Moazzami","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Mahtab","middleName":"","lastName":"Moazzami","suffix":""},{"id":580439617,"identity":"9452ed45-0a32-448a-b2a8-cb3d4d659150","order_by":2,"name":"Nader Nokhodchi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAvklEQVRIiWNgGAWjYFACNjYgZkhgY29gg/CJ18JzgFQtDBIJRCgGAXP2trQHH8rs8vgk35g9YKixY+CTPoBfi2XPseOGM84lF7NJ55gbMBxLZmDjS8CvxeBGeps0b9uBxDbpHDMJBrYDDGw8BByG0CJ5BqjlH1Fa0o5BtEjwmEkwthGj5cyxNEmgXxLbeNLKDRL7knkIazneZiYBDLHE+e2Htz348M1OTr6HgBZUkMDAQMiOUTAKRsEoGAXEAACRczg+kIuKjQAAAABJRU5ErkJggg==","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Nader","middleName":"","lastName":"Nokhodchi","suffix":""},{"id":580439622,"identity":"fe46b7c9-ddce-4b9d-a077-798272db9224","order_by":3,"name":"Hossein TaheriChadorneshin","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Hossein","middleName":"","lastName":"TaheriChadorneshin","suffix":""}],"badges":[],"createdAt":"2025-04-21 07:53:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6493746/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6493746/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101244208,"identity":"e7bdb692-add7-453c-8a7e-155d298c3c70","added_by":"auto","created_at":"2026-01-27 16:17:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32721,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMRC test steps\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6493746/v1/b6ac0c121b958c06076d8c52.png"},{"id":106613534,"identity":"268c6ee8-3999-46aa-937d-679776cf38aa","added_by":"auto","created_at":"2026-04-10 12:42:18","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":756503,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6493746/v1/25fb4b87-2365-4b91-9528-53a21e1171a2.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Changes in Leptin Gene Expression and Receptor in Aged Male Rats Following Continuous Training with and without Calorie Restriction","fulltext":[{"header":"Introduction","content":"\u003cp\u003eCardiovascular disease (CVD) is currently the leading cause of death worldwide. CVD-induced deaths increased from 17.8 to 18.6 million between 2017 and 2019, reflecting a significant rise in mortality associated with this condition. This alarming increase highlights the growing impact of CVD on global health [1, 2].\u003c/p\u003e\n\u003cp\u003eAtherosclerosis, or coronary artery disease, is one of the most prominent forms of CVD. It is a complex clinical condition characterized by an inflammatory response to vascular injury. The injury causes endothelial dysfunction in the early stages of atherosclerosis. Over time, the accumulation and transformation of lipids, inflammatory cells, smooth muscle cells, and necrotic cell debris in the intima layer (beneath the single layer of endothelial cells) lead to the formation of fatty streaks, plaque buildup, increased vulnerability of blood vessels, and eventually, vascular rupture [3, 4]. The primary component of this disease is the accumulation of fat and inflammation in the large arteries, which can reduce blood flow in the lumen by more than 50%. Ultimately, it may lead to clinical complications such as myocardial infarction, ischemic cardiomyopathy (the most common cause of heart failure), and stroke [5, 6].\u003c/p\u003e\n\u003cp\u003eAtherosclerosis, predominantly a disease of older adults, is exacerbated by increased life expectancy and a growing aging population [7]. Factors such as cellular senescence [8], weight gain (particularly the accumulation of visceral fat) in old age [9], and insufficient physical activity—falling short of the levels recommended by the World Health Organization for many elderly individuals—raise significant concerns regarding the future burden of CVD [10]. Consequently, there is a pressing need to focus on the pathology of atherosclerosis and explore effective prevention strategies.\u003c/p\u003e\n\u003cp\u003eIn general, advancing age, obesity, and a sedentary lifestyle are the most critical factors contributing to atherosclerosis [11]. Reportedly, a 10 kg increase in body weight raises the risk of coronary artery disease by 12% while also increasing systolic blood pressure by approximately 3 mm Hg and diastolic blood pressure by 2.3 mm Hg [12]. The link between obesity and cardiovascular risk is primarily driven by the increased body fat percentage that accompanies weight gain. Adipose tissue, a highly active and metabolically significant endocrine organ, secretes biologically active molecules known as adipokines or adipocytokines. Among these, important adipokines that are elevated in visceral obesity include leptin, resistin, retinol-binding protein-4, angiopoietin-like protein 2 (AngptL-2), interleukin-6, and Monocyte Chemotactic Protein-1 (MCP-1). These adipokines contribute to systemic inflammation, endothelial dysfunction, hypercoagulability, and insulin resistance, all of which are key contributors to atherosclerosis [13]. Among these adipokines, however, leptin's specific role in atherosclerosis development remains ambiguous.\u003c/p\u003e\n\u003cp\u003eLeptin, a 16 kDa peptide hormone produced by the LEP gene, is primarily secreted from adipose tissue. Leptin binding to its specific receptor (LEPR) in the arcuate nucleus of the hypothalamus regulates food intake, energy expenditure, and hormone secretion. Therefore, leptin plays a crucial role in regulating body weight, energy balance, inflammatory responses, immune function, angiogenesis, vascular function, bone homeostasis, and reproduction [14]. Circulating leptin levels (typically around 16 ng/ml) reflect the amount of energy stored in adipose tissue and are directly related to the degree of obesity. Increased adipose tissue may lead to leptin resistance and elevated leptin levels [15]. Various factors, including inflammatory cytokines, glucocorticoids, and insulin, regulate the expression and secretion of leptin. Conversely, sympathetic norepinephrine release and β-adrenergic receptor activation in adipose tissue are key factors in reducing leptin gene expression in response to leptin injection [16].\u003c/p\u003e\n\u003cp\u003eNotably, leptin is too large to cross the blood-brain barrier passively; hence, it is transported through a saturable and regulated transport system. While leptin reaches most central LepRb neurons via this system or through the cerebrospinal fluid, arcuate nucleus LepRb neurons can uniquely detect changes in circulating leptin independent of the blood-brain barrier, responding to these changes with time- and dose-dependent sensitivity [16]. Previous studies have indicated that patients with CVD exhibit elevated serum leptin levels compared to healthy individuals [17]. These findings indicate that leptin may facilitate an inflammatory state that enhances monocyte recruitment to the arterial intima and stimulates pro-inflammatory cytokine production, thereby positioning leptin as a potential mediator of atherosclerosis [18]. Additionally, leptin can induce a prothrombotic state by increasing the expression of Plasminogen Activator Inhibitor-1 (PAI-1) a major endogenous inhibitor of fibrinolysis [19]. More importantly, age-related loss of leptin responsiveness in stress signaling and eNOS phosphorylation may cause changes in cardiac geometry and function, particularly in obese and older people [20].\u003c/p\u003e\n\u003cp\u003eThere are six leptin receptor (LepR) isoforms, all of which are alternative splicing products of the db gene transcript. These receptors exhibit different expression patterns across various tissues. Although they share identical extracellular and membrane-spanning domains, their intracellular domains differ in length. Type 2 leptin receptors in peripheral tissues play a significant role in regulating energy and body weight in these tissues upon activation [21].\u003c/p\u003e\n\u003cp\u003eLifestyle interventions, such as low-calorie diets and regular exercise, are commonly recommended as primary strategies to improve the health of individuals at risk for atherosclerosis\u0026nbsp;[22]. The combination of dietary and exercise interventions is expected to have additive effects. Meta-analyses in this field suggest that combining exercise training with calorie restriction may be more effective than either intervention alone [23, 24]. However, these results are not always consistent. For instance, a recently published meta-analysis did not find significant reductions in inflammatory marker concentrations following concurrent diet and exercise interventions compared to diet alone [25].\u003c/p\u003e\n\u003cp\u003eThe conflicting results of these studies highlight the need for further research with more controlled variables. Therefore, this research aimed to investigate and compare the changes in serum levels of leptin, leptin receptor, and PAI-1 following a period of continuous training with and without calorie restriction in aged male rats.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cp\u003e\u003cstrong\u003eExperimental Animals\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis experiment was approved by the Laboratory Animal Ethics Committee of \u0026nbsp;Bojnord University. The study involved 24 male Wistar rats with a mean age of 26 months and an average weight of 200\u0026ndash;250 grams. The rats were purchased from the Institute Pasteur of Tehran. In order to ensure proper acclimatization, all rats were exposed to a 12-hour light/12-hour dark cycle, maintained at an ambient temperature of 23\u0026ndash;25\u0026deg;C and an ambient humidity of 55\u0026ndash;65%.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental Design and Diet\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe statistical sample was randomly assigned to four groups, each consisting of six rats: continuous training, continuous training with calorie restriction, calorie restriction, and control. The maximum running capacity (MRC) test was performed to assess the aerobic performance of the rats. The test was conducted until the point of fatigue, i.e., after two stimulations with a soft shoulder and no response from the rat. The general outline of this test is depicted in Figure 1.\u003c/p\u003e\n\u003cp\u003eThis test was conducted at baseline and the end of the study. The average speed in the continuous training group and continuous training with calorie restriction group was 35.66 m/min and 37.33 m/min, respectively. Moreover, the average test duration was 21.16 minutes and 17.26 minutes, respectively.\u003c/p\u003e\n\u003cp\u003eDuring the study, each rat underwent a six-week training program based on its group assignment, consisting of five 50-minute sessions per week. During the first two weeks, the rats were familiarized with the rodent-specific treadmill by exercising for 5 to 10 minutes at a speed of 5 to 10 meters per minute. Each training session included a 5-minute warm-up and cool-down period at an intensity of 30% of the maximum speed (10 meters per minute).\u003c/p\u003e\n\u003cp\u003eThe continuous training started with 10 minutes per session at an intensity of 60% of the maximum speed (20 meters per minute) during the first week. It gradually increased to 40 minutes per session by the sixth week while maintaining the same intensity. Maximum speed during maximum oxygen consumption was determined using the test protocol by Leonardo et al. (2007) to establish training intensity. This protocol involved a 3-minute warm-up at a speed of 5 meters per minute, after which the treadmill speed was increased by 4 meters per minute every two minutes. The maximum speed was defined as the point where laboratory rats could no longer maintain a steady pace for at least 30 seconds and failed to continue running despite an increase in speed\u003cspan dir=\"RTL\"\u003e\u0026nbsp;[26]\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eThe required pelleted feed for the rats was purchased from the knowledge-based company Med Zist Samaneh Pishro. The amount of food provided to the rats, excluding the calorie-restricted groups, was 10 grams of pellets per 100 grams of body weight, along with 10 to 15 milliliters of water. However, the calorie-restricted groups received 30% less food than the ad libitum amount. The percentage of food reduction was adjusted based on subsequent weight measurements, ultimately reaching a 20% reduction in food intake.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRat Dissection and Tissue Sampling\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eForty-eight hours after the final training session and an 8-hour fast, the animals were anesthetized using a combination of ketamine (90 mg/kg) and xylazine (10 mg/kg). The abdominal cavity was subsequently opened, and upon locating the aorta, a suitably sized section was excised. Additionally, a portion of visceral fat was collected, immediately placed in a labeled 1.8 mL microtube, and rapidly frozen in liquid nitrogen before it was stored at -80\u0026deg;C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ecDNA Synthesis for PCR (Reverse-Transcriptase) Reaction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe reverse-transcriptase PCR (RT-PCR) is one of the most sensitive methods for detecting RNA. The kit used for cDNA synthesis provided a complete system for synthesizing the first complementary strand of cDNA from RNA, capable of synthesizing cDNA up to 13 kilobases in length. The kit included both oligo(dT) and random hexamer primers. In order to verify the accuracy of cDNA synthesis, a PCR reaction was conducted using primers specific to the GPI control gene. The PCR product was subsequently visualized on an agarose gel, where clear bands were observed at approximately 350 base pairs for the GPI gene, with no secondary bands present, confirming the quality of the synthesized cDNA.\u003c/p\u003e\n\u003cp\u003eAfter cDNA synthesis, the quality of each extracted sample was measured using a nanodrop spectrophotometer at a wavelength of 260 nm. All samples had concentrations exceeding 450 ng/\u0026mu;L.\u003c/p\u003e\n\u003cp\u003ePrimer Design for RQ-PCR (Relative Quantitation) Reaction\u003c/p\u003e\n\u003cp\u003eOnce the nucleotide sequence for the target gene was acquired, BLAST software was used on the NCBI website, as per the research by Baldus et al., to ascertain primer specificity and evaluate the homology of the primer sequence with registered genetic sequences.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Sequence of primers\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGene name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eprimers\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSequence\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL49+8fO5RJGpg8ebLuxMWLQ6Bc4kF9fb0M2bZ2zpgxC8ocBaNgFIyCUUAMYGAAACVWDgDyW4e3AAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/G5RJGpg2bZr6hIULvaBc4kF9fb0E2bZ2zJjRA2WOglEwCkbBKCAGMDAAAJQBDeUk5x4hAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAA+SURBVEhL7c2hDQAgDETRkpDgmIFNK9FVDNBBi+kABf2f+qdOAABPIqJlvjGzYe4zZ52q9u/Xfc7KBABUiFxn/wzyvBKH9AAAAABJRU5ErkJggg==\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/C5RJGpg+fbr05IULTaBc4kF9fb0A2ba2z5iRCWWOglEwCkbBKCAGMDAAAPb7DZmD6kIsAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAA/SURBVEhL7c2hEQAgDATBzGCx9EBB9BOLDQZJqcGkgIC+VffqBQDwxN1L5Bsza+ucHjNPVev369x7RAIAMkQueikNVHTrBjYAAAAASUVORK5CYII=\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/C5RJGpg+fbr05IULTaBc4kF9fb0A2ba2z5iRCWWOglEwCkbBKCAGMDAAAPb7DZmD6kIsAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003c/p\u003e\n \u003cp\u003ePAI-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/C5RJGpg+fbr05IULTaBc4kF9fb0A2ba2z5iRCWWOglEwCkbBKCAGMDAAAPb7DZmD6kIsAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAzCAYAAACnrDmiAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/I5RJGpg4cSL7xMWL+aBc4kF9fT0L2ba2T5+uAGWOglEwCkbBKKApYGAAAEoODPKRhfeDAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"51\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAA+SURBVEhL7c2hDQAgDETRkpDgmIFNK9FVDNBBi+kABf2f+qdOAABPIqJlvjGzYe4zZ52q9u/Xfc7KBABUiFxn/wzyvBKH9AAAAABJRU5ErkJggg==\" alt=\"image\" width=\"14\" height=\"50\"\u003eForward\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eTGAGAGAGGGCAAAGTGGTT\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eACGTGTCCACTGCTCACTTA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eLeptin\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAA+SURBVEhL7c2hDQAgDETRkpDgmIFNK9FVDNBBi+kABf2f+qdOAABPIqJlvjGzYe4zZ52q9u/Xfc7KBABUiFxn/wzyvBKH9AAAAABJRU5ErkJggg==\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/C5RJGpg+fbr05IULTaBc4kF9fb0A2ba2z5iRCWWOglEwCkbBKCAGMDAAAPb7DZmD6kIsAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"50\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAzCAYAAACnrDmiAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAABASURBVEhLYxgFo2AUjIJRQBL4//8/I5RJGpg4cSL7xMWL+aBc4kF9fT0L2ba2T5+uAGWOglEwCkbBKKApYGAAAEoODPKRhfeDAAAAAElFTkSuQmCC\" alt=\"image\" width=\"14\" height=\"51\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAA4AAAAyCAYAAABs8OoHAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAA+SURBVEhL7c2hDQAgDETRkpDgmIFNK9FVDNBBi+kABf2f+qdOAABPIqJlvjGzYe4zZ52q9u/Xfc7KBABUiFxn/wzyvBKH9AAAAABJRU5ErkJggg==\" alt=\"image\" width=\"14\" height=\"50\"\u003eForward\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGAC ATT TCA CAC ACG CAG TC\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGAG GAG GTC TCG CAG GTT\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eLeptin receptor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eForward\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCAC TGT TAA TTT CAC ACC AGA G\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGTC ATT CAA ACC ATA GTT TAG G\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003ch3\u003eReal-Time PCR Analysis\u003c/h3\u003e\n\u003cp\u003eReal-time PCR was conducted using an ABI thermocycler. The PCR reactions were performed after the primers had been prepared. The \u0026Delta;CT values for the Aplin, APJ, and ACE2 genes were calculated for both the control (healthy) and experimental samples. Expression levels were assessed by calculating the relative quantity (Fold Change) using the appropriate formula. Data were analyzed using one-way analysis of variance (ANOVA) with SPSS software version 26.\u003c/p\u003e\n\u003ch3\u003eStatistical Results\u003c/h3\u003e\n\u003cp\u003eInitially, the height and weight data of the rats were analyzed using descriptive statistics, including mean, standard deviation, minimum, and maximum (Table 2).\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"746\" height=\"183\"\u003e\u003c/p\u003e\n\u003cp\u003eThe normality of the research variables was assessed using the Shapiro-Wilk and Lune tests. The results indicated that leptin levels and leptin receptor expression did not follow a normal distribution, while PAI-1 levels were normally distributed. Consequently, the Kruskal-Wallis test, a non-parametric method, was employed to analyze leptin and leptin receptor data, whereas one-way ANOVA as a parametric test was used for the PAI-1 data. Table 3 presents the changes in data related to leptin and its receptor.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Kruskal-Wallis test results for leptin and leptin receptor\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eVariables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eStatistic\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSig\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003eLeptin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e12.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e18.274\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eContinuous Exercise with Calorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003eLeptin receptor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.156\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e12.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\"\u003e\n \u003cp\u003e*0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eContinuous Exercise with Calorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.54\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.97\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe results of the Kruskal-Wallis test revealed that the significance level for both leptin levels and leptin receptor expression was less than 0.05, indicating significant changes in these variables. Moreover, Bonferroni\u0026apos;s post hoc test was employed to identify the specific locations of these differences. The findings from this post hoc analysis are presented in Table 4.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. Bonferroni post hoc test results\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eVariables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups 1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups 2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSig\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"6\"\u003e\n \u003cp\u003eLeptin\u003c/p\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise with Calorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e9.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e5.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.039\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e11.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e4.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.494\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.012\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"6\"\u003e\n \u003cp\u003eLeptin receptor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise with Calorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e6.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.069\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e*0.002\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.017\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eContinuous Exercise\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.100\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.496\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCalorie Restriction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e0.315\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003cp\u003eAdditionally, in order to investigate the changes in PAI-1 expression in the aorta tissue of rats, a one-way analysis of variance test was used, as displayed in Table 5.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5. One-way ANOVA results of PAI-1 expression\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSource of changes\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSum of squares\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMean square\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eSig\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBetween-group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.908\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.303\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\"\u003e\n \u003cp\u003e1.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\"\u003e\n \u003cp\u003e0.254\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eWithin-group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.206\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.034\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe results of one-way ANOVA indicated that the significance level for PAI-1 levels was greater than 0.05, suggesting the absence of any significant changes in this variable.\u003c/p\u003e"},{"header":"Discussion and Conclusion","content":"\u003cp\u003eOver the past decade, numerous studies have explored the diverse roles of leptin and its potential effects on the development or exacerbation of conditions such as obesity, diabetes, metabolic syndrome, obstructive pulmonary disease, bone density issues, and CVD. These studies aim to mitigate the spread of these diseases and improve lifestyle quality. The present study aligns with previous research, seeking to determine whether combining exercise and calorie restriction is more effective than implementing either intervention alone.\u003c/p\u003e\n\u003cp\u003eIn summary, the results of this study demonstrate a significant difference in the effects of continuous exercise combined with calorie restriction versus either intervention alone on leptin gene expression in aged male rats. Specifically, both calorie restriction and continuous exercise with calorie restriction significantly increased leptin gene expression in these rats. Additionally, a significant difference was observed between the effects of continuous training with and without calorie restriction on leptin receptor gene expression. Indeed, continuous training combined with calorie restriction significantly reduced leptin receptor gene expression. Finally, no significant difference was found between the effects of continuous exercise with and without calorie restriction on PAI-1 expression in the aortic tissue of aged male rats.\u003c/p\u003e\n\u003cp\u003eThese findings contrast with previous reports, such as the study by Tremblay et al. (2019), which described decreased leptin levels following combined interventions [27]. A critical factor to consider is that, in obese populations, the high volume of adipose tissue and subsequent elevated secretion of adipokines often result in leptin resistance. Under these conditions, the hypothalamus becomes less responsive to leptin, leading to persistent hunger and unchanged food intake despite the abundant energy stored as fat [28]. In such cases, regular exercise can reduce adipose tissue and, consequently, decrease adipokine secretion, leading to lower leptin levels. It has been suggested that the adipokine response to exercise is correlated with the amount of adipose tissue [29].\u003c/p\u003e\n\u003cp\u003eHowever, in populations with normal adipose tissue levels, the significant increase in leptin levels observed after combined interventions might be attributed to the metabolic demands imposed by reduced caloric intake and increased energy expenditure due to physical activity. Under conditions that elevate the metabolic rate, such as exercise, leptin secretion by adipocytes gradually increases. Leptin signaling through its receptors in the skeletal muscle activates the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α). In physiological conditions, increased leptin concentration leads to higher expression of leptin receptors and enhanced leptin reabsorption, creating a positive feedback loop that strengthens the leptin signal [30].\u003c/p\u003e\n\u003cp\u003eThe enhanced effects of leptin during continuous exercise coupled with calorie restriction are noteworthy. Previous studies have indicated that weight loss efforts, regardless of whether they involve obese individuals or those without a history of obesity, frequently result in a disproportionate decrease in energy expenditure compared to the reduction in body mass. In such cases, the tone of the sympathetic nervous system and the circulating concentrations of leptin and bioactive thyroid hormones decrease. In contrast, the tone of the parasympathetic nervous system increases, ultimately resulting in unsuccessful weight loss attempts. Decreased leptin levels signal the body to restore energy balance by increasing appetite and decreasing energy expenditure [31]. Conversely, elevated leptin levels suppress appetite by reducing hypothalamic neuropeptide Y (an appetite stimulator) and increasing α-MSH (an appetite suppressant), as well as by enhancing energy expenditure through increased secretion of TRH and GnRH [32]. In this context, it has been shown that lean mice receiving chronic leptin injections lose fat mass in a dose-dependent manner (within the physiological range). Leptin treatment has also been reported to reverse the changes in circulating thyroid hormone and corticosterone levels that are typically associated with food deprivation [33].\u003c/p\u003e\n\u003cp\u003eThis natural increase in leptin levels can have beneficial effects on atherosclerosis. Leptin treatment appears to reduce vascular inflammation and endothelial-to-mesenchymal transition, a key factor in atherosclerosis development. Previous research has demonstrated that leptin treatment of endothelial cells significantly increases the expression of pro-inflammatory markers, such as intercellular adhesion molecule-1 and E-selectin, at the mRNA level. This may suggest that leptin mediates anti-inflammatory properties in the endothelium [34].\u003c/p\u003e\n\u003cp\u003eMoreover, plasma leptin levels appear to be inversely correlated with ObR expression in the arcuate nucleus of the hypothalamus, consistent with the changes in leptin receptor levels observed in the present study [35]. While previous studies suggest that increased leptin levels could be a direct outcome of exercise, the results of this research clearly show that combined interventions can have synergistic effects [36]. It has been proposed that when leptin binds to its soluble receptor, it is unable to activate membrane receptors, leading to the hypothesis that the soluble form of leptin receptors may be involved in leptin resistance [36].\u003c/p\u003e\n\u003cp\u003eAdditionally, the results indicated no significant difference between the effects of continuous training with and without calorie restriction on PAI-1 expression in the aortic tissue of aged male rats. Within-group analysis showed increased, though insignificant, PAI-1 gene expression following exercise and calorie restriction. These findings align with previous studies reporting improvements in PAI-1 fibrinolytic activity following aerobic exercise [37]. However, the absence of a significant difference in the current study may be attributed to variations in baseline PAI-1 levels, which impedes drawing definitive conclusions. The small sample size may further account for this outcome; therefore, additional research is recommended to explore this issue in more depth.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCompliance with Ethical Standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare there no conflict of interest. All procedures involving animals were carried out in compliance with the relevant national regulations, including the Iranian Animal Welfare Law, and the ethical guidelines set by the Bioethics Committee of Birjand University, under approval number [IR.UB.REC.1401.010].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received no external funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed equally to the preparation of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research project was conducted in the Animal Laboratory of the Faculty of Physical Education at Ferdowsi University of Mashhad. We would like to express our sincere gratitude to the faculty members and staff of the department for their valuable cooperation and support, particularly in facilitating access to the laboratory facilities.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eTownsend, N., et al., \u003cem\u003eEpidemiology of cardiovascular disease in Europe.\u003c/em\u003e Nature Reviews Cardiology, 2022. 19(2): p. 133-143.\u003c/li\u003e\n\u003cli\u003eAlonso, A., et al., \u003cem\u003eHeart disease and stroke statistics\u0026mdash;2021 update.\u003c/em\u003e Circulation, 2021. 2021(143): p. e00-e00.\u003c/li\u003e\n\u003cli\u003ePark, J.B. and A. Avolio, \u003cem\u003eArteriosclerosis and atherosclerosis assessment in clinical practice: Methods and significance.\u003c/em\u003e Pulse, 2023. 11(1): p. 1-8.\u003c/li\u003e\n\u003cli\u003eBj\u0026ouml;rkegren, J.L.M. and A.J. 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Heeren, \u003cem\u003eThe endocrine function of adipose tissues in health and cardiometabolic disease.\u003c/em\u003e Nature reviews endocrinology, 2019. 15(9): p. 507-524.\u003c/li\u003e\n\u003cli\u003eSingh, P., et al., \u003cem\u003eLeptin upregulates the expression of plasminogen activator inhibitor-1 in human vascular endothelial cells.\u003c/em\u003e Biochemical and biophysical research communications, 2010. 392(1): p. 47-52.\u003c/li\u003e\n\u003cli\u003eJin, W., et al., \u003cem\u003eInterplay between obesity and aging on myocardial geometry and function: role of leptin-STAT3-stress signaling.\u003c/em\u003e Biochimica et Biophysica Acta (BBA)-General Subjects, 2023. 1867(2): p. 130281.\u003c/li\u003e\n\u003cli\u003eSurmi, B.K., et al., \u003cem\u003eThe role of macrophage leptin receptor in aortic root lesion formation.\u003c/em\u003e American Journal of Physiology-Endocrinology and Metabolism, 2008. 294(3): p. E488-E495.\u003c/li\u003e\n\u003cli\u003eBorges, J.H., et al., \u003cem\u003eExercise training and/or diet on reduction of intra-abdominal adipose tissue and risk factors for cardiovascular disease.\u003c/em\u003e European journal of clinical nutrition, 2019. 73(7): p. 1063-1068.\u003c/li\u003e\n\u003cli\u003eKhalafi, M., M.E. Symonds, and A. 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Murawska-Ciałowicz, \u003cem\u003eExercise and Weight Management: The Role of Leptin\u0026mdash;A Systematic Review and Update of Clinical Data from 2000\u0026ndash;2022.\u003c/em\u003e Journal of Clinical Medicine, 2023. 12(13): p. 4490.\u003c/li\u003e\n\u003cli\u003eRosenbaum, M., et al., \u003cem\u003eLow-dose leptin reverses skeletal muscle, autonomic, and neuroendocrine adaptations to maintenance of reduced weight.\u003c/em\u003e The Journal of clinical investigation, 2005. 115(12): p. 3579-3586.\u003c/li\u003e\n\u003cli\u003eIzquierdo, A.G., et al., \u003cem\u003eLeptin, obesity, and leptin resistance: where are we 25 years later?\u003c/em\u003e Nutrients, 2019. 11(11): p. 2704.\u003c/li\u003e\n\u003cli\u003eFriedman, J.M., \u003cem\u003eThe function of leptin in nutrition, weight, and physiology.\u003c/em\u003e Nutrition reviews, 2002. 60(suppl_10): p. S1-S14.\u003c/li\u003e\n\u003cli\u003eSt\u0026uuml;rzebecher, P.E., et al., \u003cem\u003eLeptin treatment has vasculo-protective effects in lipodystrophic mice.\u003c/em\u003e Proceedings of the National Academy of Sciences, 2022. 119(40): p. e2110374119.\u003c/li\u003e\n\u003cli\u003eFernandez-Galaz, C., et al., \u003cem\u003eLong-term food restriction prevents ageing-associated central leptin resistance in wistar rats.\u003c/em\u003e Diabetologia, 2002. 45: p. 997-1003.\u003c/li\u003e\n\u003cli\u003eYasari, S., et al., \u003cem\u003eExercise training decreases plasma leptin levels and the expression of hepatic leptin receptor-a,-b, and,-e in rats.\u003c/em\u003e Molecular and Cellular Biochemistry, 2009. 324: p. 13-20.\u003c/li\u003e\n\u003cli\u003eHabibian, M. and S.A. Rezaye Mousavi, \u003cem\u003eThe Effects of Regular Aerobic Exercise on the Serum levels of Plasminogen Activator Inhibitor-1 and Adiponectin in Obese Girls.\u003c/em\u003e Journal of Inflammatory Diseases, 2019. 23(5): p. 418-429.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Leptin, Continuous Training, Calorie Restriction, atherosclerosis","lastPublishedDoi":"10.21203/rs.3.rs-6493746/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6493746/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose\u003c/strong\u003e: The present study aimed to examine the effects of continuous training with and without calorie restriction on atherosclerosis indices in aged male rats.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: this experimental study was conducted with a posttest design and a control group. The subjects consisted of 24 aged male rats, purposefully assigned to four groups based on weight: control, calorie restriction, continuous exercise, and continuous exercise with calorie restriction. The training duration was six weeks, comprising five 50-minute sessions per week, totaling 30 sessions. Each session included three phases: warm-up, cool-down (each featuring 6–8 m/min), and the main exercise (continuous running at 10 m/min on a specialized treadmill). The calorie restriction program involved reducing food intake by 30%. 48 hours after the last training session and an 8-hour fast, the rats were anesthetized with a combination of ketamine (90 mg/kg) and xylazine (10 mg/kg).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: continuous training combined with calorie restriction significantly affected leptin gene expression and its receptor in aged male rats (P\u0026lt;0.05). However, no significant differences were observed between the continuous training groups, with and without calorie restriction, regarding the expression of the PAI-1 gene in aged male rats (P\u0026gt;0.05).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: These findings demonstrate that continuous exercise and calorie restriction can influence leptin gene expression and its receptor.\u003c/p\u003e","manuscriptTitle":"Changes in Leptin Gene Expression and Receptor in Aged Male Rats Following Continuous Training with and without Calorie Restriction","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-27 16:17:05","doi":"10.21203/rs.3.rs-6493746/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8b949b93-f97d-4d89-ae5c-c9cd2ad235d1","owner":[],"postedDate":"January 27th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-10T12:40:40+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-27 16:17:05","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6493746","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6493746","identity":"rs-6493746","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}