The aim of this study was to determine the effects of resistance training on the immunologic response, body composition, tumor necrosis factor-alpha (TNF-alpha) gene expression obtained from blood leukocytes, and the cytokines interleukin-6, TNF-alpha, and C-reactive protein (CRP), in the elderly women (mean age 63 ± 2 y). A randomized controlled trial was performed using a bi-set training method for eight weeks in nineteen elderly women. Peripheral blood samples were collected by puncture in pretraining (Pre) and posttraining (Post) moments. In the resistance training group, there was a statistically significant decrease from 38.43 ± 9.48 pg/mL to 11.76 ± 5.19 pg/mL (
In recent years, life expectancy has grown considerably. Nowadays, there are 14.3 million Brazilians over 60 years. Until the year 2025, Brazil will become the sixth country in the number of elderly people [
Human aging is a process characterized by biological and physiological changes with repercussion on body composition and onset of chronic systemic inflammation, pointed by the increase of inflammatory markers such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-
Cytokines are proteins that respond to pathogens with cells of innate and adaptive immunity and also act on inflammatory processes [
Another important cytokine in the inflammatory process is IL-6. It is synthesized from mononuclear phagocytes, vascular endothelial cells, fibroblasts, and adipose and muscle tissues. Its function is stimulation of neutrophil production by bone marrow progenitors, promoting synthesis of proinflammatory cytokines and inhibiting TNF-
C-reactive protein is a risk marker for cardiovascular disease, which also features an important function in the inflammatory process, especially in the acute phase, when CRP can increase up to 1000 times its normal serum level. This protein is produced and released into the bloodstream by hepatocytes, but it can also be produced by arterial walls. Its synthesis is stimulated by IL-1 and IL-6, which are released by macrophages [
Several systemic, tissue, cellular, and molecular changes happen in the aging process, featured by progressive dysfunction of the immune system, with an imbalance in the concentrations of pro- and anti-inflammatory cytokines and other inflammatory markers associated with the development of cardiovascular diseases, and the increase in morbidity and mortality, in the elderly [
Therefore, the role of immunological and inflammatory markers such as TNF-
Therefore, the role of immunological and inflammatory markers such as TNF-
It is an experimental and controlled study. The Ethics and Research Committee of the CEUMA University, through the Brazil Platform Ministry of Health, National Health Council, National Commission of Ethics in Research-Division of Experimental Research Involving Human Beings, approved this study under CAEE permit number 10863313.2.1001.5084 (protocol no. 372453/2013). All participants signed an individual consent form, so their identification is included in this article. All procedures performed in this study were in accordance with the Ethical Standards of the Institutional and/or National Research Committee and with the Helsinki Declaration of 1975 and its later amendments or comparable ethical standards.
This is a nonprobabilistic sample. A list with names and telephone contacts of women aged between 60 and 70 was initially requested from the Integrated Seniors University (UNITI/UFMA). From the telephone contacts available, subjects were invited to take part in the study, and those who had not entered any other structured RT program in the last six months were selected. A meeting was held for explanation about project logistics, for delivering the schedule of activities to all participants, and for the consent form to be signed by them. Also, there were scheduled days, times, and locations for anthropometric evaluation and blood collection. For inclusion criteria, participants should be between 60 and 70 years; not be enrolled in structured RT programs in the last six months; be nonsmokers; have no degree of obesity according to the World Health Organization [
A pretest/posttest with a control group design was adopted for the study. Both groups were pretested and posttested at eight weeks. The participants were randomly (
Study design flowchart.
The RT protocol was based on a critical analysis of the databases in [
For the protocol procedure, before and after all training sessions, participants rested for 5 minutes and then blood pressure was measured. The RT was supervised by one physiotherapist and two physical education professionals. The model of resistance training was by means of 8–12 maximal repetitions (MRs), aiming at muscular hypertrophy. In relation to the weekly frequency, the training protocol was performed three times per week, where eight exercises were performed in LL and UL: sitting leg press, sitting supine, knee extension, pulley (back), lying down knee flexion, low pulley elbow flexion, seated leg press, and pulley elbow extension. As for the increase in intensity (load (kg)), two criteria were adopted: (1) exercise intensity and load increase were obtained using the BORG scale; in other words, participants were instructed to mention any number from this scale, provided that the number actually corresponded to their perceived effort after each exercise series was performed. (2) After that, all participants performed their RT program within a maximum repetition (MR) zone of 8 to 12 repetitions; thus, whenever the participants exceeded the limits of this zone, a new increase of load occurred which would remain within the established zone, corresponding to 15% of the initial load. The time interval between the repetitions was one minute for each series, and the duration of the training protocol sessions lasted on average 50 minutes. In relation to the speed of execution of the exercise preestablished in four seconds for each movement, the repetition rate was performed with a concentric action of 2 seconds and an eccentric action of 2 seconds, controlled by visual and verbal commands to standardize angles of movement. The intervention of resistance training was very intense, and the participants were elderly and had not trained for at least six months, so in order to avoid possible side effects and possible withdrawal of the participants in the research, we chose to do a one-week adaptation training in the participants performed two sets of 15 submaximal repetitions with low loads, aiming at neuromuscular adaptation avoiding late-onset muscular pain.
All participants were submitted to anthropometric assessment in pretest and posttest (eight weeks later). Body mass was measured using a digital scale (Welmy® W300, Brazil) with a maximum capacity of 300 kg. Height was measured using an anthropometric scale between 1.00 and 2.00 m. These measurements were used to calculate the body mass index (BMI). Also, waist and hip circumference was measured using the metric tape (Waist Fit®, Brazil) to calculate the waist-to-hip ratio. Prior to the measurements of body composition, participants were instructed to not eat 2-3 hours before, not drink alcohol, not perform any physical exercise for 24 hours, monitor fluid intake, and urinate 30 minutes prior to assessment. To begin the assessment, the participants were asked to lie down on the hammock, and then electrodes were put at predetermined points which were sanitized using 70% alcohol.
The emitter electrodes were placed at the dorsal surface of the right hand, near the metacarpophalangeal joint, and at the transverse arch of the upper surface of the right foot. The detector electrodes were placed between the distal prominences of the radius and ulna of the right wrist, and another one was placed between the medial malleolus and lateral malleolus of the right ankle [
Blood samples were collected by a trained pharmaceutical and in accordance with the biosafety standards recommended by NR32. The blood collection was performed at Pre (fasting) and Post (24 hours after the last day of RT) moments. At the moment Pre, all participants were instructed to go to the Laboratory of Physiology and Exercise Prescription (LAFIPEMA-UFMA), at 6 am, fasted for 8–12 hours. At the time Post, participants in the Control group made blood collections after a time period of 8 weeks. Already, the participants who performed RT made blood collections 24 hours after the last day of training. A total volume of 14 ml, approximately, was collected at vacuum and then distributed into a 4 ml EDTA tube (Vacuette) and two dry tubes containing 5 ml separating gel (Vacuette). The samples collected proceeded with the following usage: (a) the tube containing the EDTA anticoagulant was used to perform the complete blood count and the analysis of TNF-
This was analyzed to measure the CRP serum levels and feature the degree of risk for cardiovascular diseases. Therefore, biochemical analysis was performed by immunoturbidimetry.
The analysis of the blood concentrations of IL-6 and TNF-
All concentrations of IL-6 and TNF-
The RNA isolation was performed from blood leukocytes collected in a vacuum tube containing the EDTA anticoagulant. A sample volume of 20
Real-time quantitative PCR assays were performed for TNF-
The statistical analysis was performed using the software GraphPad Prism 6.0. First, the Shapiro–Wilk normality test was performed, considering parametric tests (
Both groups presented similar variables, except for height (
Characteristics of study participants.
Variable | Control ( |
RT ( |
|
---|---|---|---|
Age (years) | 63.0 ± 1.0 | 63.0 ± 2.0 | 0.08 |
Height (m) | 1.4 ± 0.0 | 1.5 ± 0.0 | 0.01 |
Body mass (kg) | 58.5 ± 3.8 | 63.7 ± 2.4 | 0.04 |
Fat percentage (%) | 39.1 ± 2.6 | 39.3 ± 1.4 | 0.60 |
Lean mass percentage (%) | 60.8 ± 2.6 | 60.7 ± 1.4 | 0.60 |
Data are presented as mean ± standard deviation. Student’s
Comparing Pre and Post moments for RT and Control groups, there was a statistically significant difference in the RT group for the variables: decreased fat mass (
Anthropometric characteristics, body composition and lipid profile, in RT and Control groups at Pre and Post (8 weeks) moments.
Variables | Control ( |
|
RT ( |
| ||
---|---|---|---|---|---|---|
Pre | Post | Pre | Post | |||
Body mass (kg) | 58.5 ± 3.8 | 58.1 ± 3.8 | 0.9 | 63.7 ± 2.4 | 65.0 ± 2.5 | 0.1 |
Fat mass (kg) | 22.8 ± 2.9 | 22.2 ± 3.0 | 0.2 | 25.0 ± 1.7 | 23.5 ± 1.7 |
0.02 |
Lean mass (kg) | 35.6 ± 1.1 | 35.7 ± 1.0 | 0.8 | 38.6 ± 3.6 | 41.4 ± 4.1 |
0.02 |
Fat percentage (%) | 39.1 ± 2.6 | 38.2 ± 2.7 | 0.9 | 39.3 ± 1.4 | 36.2 ± 1.5 |
0.01 |
Lean mass percentage (%) | 60.8 ± 2.6 | 61.7 ± 2.7 | 0.5 | 60.7 ± 1.4 | 63.7 ± 1.5 |
0.01 |
BMI (kg/m2) | 26.8 ± 1.7 | 26.0 ± 1.7 | 0.1 | 27.7 ± 0.9 | 27.5 ± 1.0 | 0.7 |
WHR (cm) | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.3 | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.1 |
Data are presented as mean ± standard deviation. Student’s
Following the load evolution at the 1st, 4th, and 8th training weeks, there was a load increase in their respective exercises with a significant difference between the weeks (
Evolution of training load in first, fourth, and 8th weeks of RT.
Exercises | RT ( |
Effect size (Δ) |
| ||
---|---|---|---|---|---|
1st week | 4th week | 8th week | |||
Seated leg press (kg) | 20.7 ± 3.8 | 33.6 ± 1.1 |
45.48 ± 1.4 |
6.3 | 0.0001 |
Elbow flexion (low pulley) (kg) | 9.0 ± 1.8 | 13.8 ± 0.3 |
17.83 ± 0.4 |
4.8 | 0.0001 |
Knee extension (kg) | 9.9 ± 0.5 | 18.4 ± 0.6 |
25.90 ± 0.8 |
28.0 | 0.0001 |
Supine seated (kg) | 7.1 ± 0.3 | 13.8 ± 0.4 |
17.96 ± 0.7 |
29.2 | 0.0001 |
Lying knee flexion (kg) | 7.9 ± 0.4 | 15.5 ± 2.0 |
17.09 ± 0.5 |
22.1 | 0.0001 |
Pulley (back) (kg) | 13.2 ± 0.5 | 21.9 ± 0.4 |
27.03 ± 0.5 |
26.4 | 0.0001 |
Plantar flexion (seated leg press) (kg) | 20.0 ± 0.7 | 33.38 ± 0.9 |
43.56 ± 1.2 |
30.9 | 0.0001 |
Elbow extension (pulley) (kg) | 8.6 ± 0.6 | 16.42 ± 0.4 |
20.20 ± 0.4 |
19.0 | 0.0001 |
Data are presented as mean ± standard deviation. One-way ANOVA test followed by Tukey’s post hoc test:
With respect to serum concentration of IL-6, a statistically significant interaction (
Dispersion measurements of interleukin-6 between Control (
With respect to serum concentration of TNF-
Dispersion measurements of tumor necrosis factor-
The molecular analysis of TNF-
Measurements of tumor necrosis factor-
With respect to serum concentration of CRP, there was a statistically significant interaction (
Dispersion measurements of C-reactive protein between Control (
This study aimed to evaluate the effects of eight weeks of RT on the immune response of IL-6 and TNF-
The objective of this study was to evaluate the behavior of the immune system of IL-6 and TNF-
In this study, eight weeks of RT significantly decreased the serum concentrations of IL-6, TNF-
Therefore, to analyze the chronic effects of physical exercise on inflammatory markers, it is essential to understand the acute effects of exercise on cytokine concentrations. Pereira et al. [
This was observed by Córdova et al. [
Searching evidence about the molecular mechanisms behind these findings, it was verified that RT produces modulatory effects on TNF-
In order to understand the control of TNF-
As well, it is important to point out that the RT group presented significant gains in lean mass and loss of fat mass. These findings may also have contributed to the suppression of TNF-
Based on these data, adipose tissue contributes to the increase in TNF-
In this research, an increase in muscle strength was also observed, evidenced by the progressive increase in training load and in muscle volume, over the eight weeks of training. According to Prestes et al. [
Mavros et al. [
The systematized and progressive RT program is developed through appropriate and specific training purposes. Among several protocols available, the bi-set method shows an efficient proposal for a well-structured and individualized RT program, which follows the RT prescription recommendations for the elderly. In view of this, it was decided to determine the training intensity by means of an RM range between 8 and 12 [
A possible limitation of this study may be related to the representativeness of the sample, since it is a limited group of women who are available to participate in institutional care research. Another limitation of the study was in relation to the elderly women who have passed th0065 menopause period, and it is known that the hormonal changes that occur in the postmenopausal period can influence the responses to the cytokines through body composition, mainly in the fat weight. In this way, we controlled this limitation with the decrease of the fat weight with the resistance training in relation to the basal values.
In summary, the results of the present study showed that eight weeks of RT caused molecular changes in TNF-
All data used to support the findings of this study are included within the article.
The authors declare that they have no conflicts of interest.
LÂMS organized and designed the experiments. LÂMS, LGLN, WRS, and AJMCRP performed the experiments. GBP, RDL, and CTM wrote the manuscript. LÂMS, LGLN, and JOBM reviewed the manuscript. LÂMS, LGLN, and FN gave approval of the final manuscript version.
This project was funded by the Federal University of Maranhão (UFMA) and Foundation for Research Support of the State of Maranhão (FAPEMA-BM-01486/13). Luís Ângelo Macêdo Santiago was an FAPEMA productivity scholarship student.