Clin Physiol Funct Imaging (2014)
doi: 10.1111/cpf.12207
Acute eccentric resistance exercise decreases matrix metalloproteinase activity in obese elderly women ~o Luiz Quagliotti Durigan3, Rita de Cassia Marqueti3,5, Dahan da Cunha Nascimento1, James Wilfred Navalta2, Joa 1 4 Ramires Alsamir Tibana , Octavio Luiz Franco , Jesser Alves de Almeida4,5, Nathalia Ferreira Camarcßo1, Ivo Vieira de Sousa Neto1 and Jonato Prestes1 1
Graduate Program on Physical Education, Catholic University of Brasilia, Brasilia, Brazil, 2Kinesiology and Nutrition Sciences, University of Nevada, Las Vegas, NV, USA, 3University of Brasilia, 4Center for Proteomic and Biochemical Analyses, Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, and 5Graduate Program in Health Sciences and Technologies, University of Brasilia, Brasilia, Brazil
Summary Correspondence Jonato Prestes, Physical Education – Catholic University of Brasilia, Q.S. 07 Lote 01 – Bloco G 71966-700 Taguatinga, Brazil E-mail:
[email protected]
Accepted for publication Received 21 June 2014; accepted 24 September 2014
Key words blood markers; eccentric contraction; elderly; inflammation; matrix metalloproteinases; obesity
The association of ageing with obesity commits elderly women and has been correlated with multiple degenerative processes, which could be occasioned by an enhancing in levels of matrix metalloproteinase-2 and metalloproteinase-9 (MMPs) as well by an cytokine unbalance that included an enhancing on interleukin-6 (IL-6). Furthermore, other factors could be also related to degenerative process, as they could be reduced by eccentric resistance exercise (ERE), which seems particularly important to initiate resistance training in obese older adults. In this view, this study aims to determinate the effects of an acute ERE session on serum MMP-2, MMP-9 and IL-6 in elderly obese women. Ten elderly obese women participated in this study and completed a 10 repetitions maximum test (10 RM) utilizing leg extension exercise. Subjects then completed an acute ERE session consisting of seven sets of 10 repetitions at 110% of 10 RM with a rest of 3 min between sets. Blood samples were collected before, immediately after, 3, 24 and 48 h following the ERE session. Zymograms were utilized to measure the MMP-2 and MMP-9 enzymes from all individuals. Moreover, IL-6 concentration was also determinated. After ERE session, MMP-2 and MMP-9 decreased, remaining significantly below baseline values after 48 h (P30% and completion of all testing procedures. Obesity was determined as recommended by the National Institute of Diabetes and Digestive and Kidney Diseases (2001), assuming a cut-off point of 30% for women. A sedentary state was determined by answers provided on the International Physical Activity Questionnaire. Women with inflammatory, rheumatic, or autoimmune conditions or use of medications (i.e. beta blockers, hormone replacement therapy, selective oestrogen receptor modulators, anti-inflammatory, insulin, fish oil and multivitamin supplements) that could modulate the biochemical response to ERE were excluded. The study was approved by the Institutional Research Ethics Committee (protocol#035/2011), and all subjects gave written informed consent and all work was conducted in accordance with the Declaration of Helsinki. 10 RM testing A ten repetition maximum (10 RM) test was performed according to Tibana et al. (2012). On the first visit, subjects completed a medical form and physical questionnaire, anthropometric measures and dual-energy X-ray absorptiometry (DXA, General Electric-GE model 8548 BX1L, 2005, Lunar DPX type, software Encore 2005, Rommelsdorf, Germany) body composition analysis and completed a familiarization session on a leg extension isoinertial machine (Righetto Fitness Equipment, Sao Paulo, Brazil) that consisted of three submaximum sets of 8–10 repetitions. The isoinertial machine was chosen for practical reasons because it is a typical human movement. In addition, knee extension exercise was chosen because it has a strong correlation with elderly survival time and dependency (Frontera et al., 2001). Three days later, subjects performed a 10 RM test, and following 72 h of rest, they completed the 10 RM test again to determine test retest reliability (R = 099). The test was terminated when voluntary concentric failure occurred (inability to perform a full range repetition of the movement as a consequence of fatigue). As described previously (Tibana et al., 2012), testing errors were minimized by the following strategies: (i) standardized instructions were given concerning all data collection procedures; (ii) exercise technique and leg extension machine adjustments were standardized for each subject; and (iii) subjects were given verbal encouragement throughout testing. Rest intervals of 3–5 min were instituted between 10 RM trials. Moreover, subjects were asked not to ingest any stimulants (e.g. caffeine) or perform any physical activity during the week prior to testing.
Methods Subjects
Acute eccentric resistance exercise
Ten elderly women from a local community (674 74 years, height 1520 02 cm, body mass 663 66 kg,
Seven days following the 10 RM tests, subjects completed an ERE protocol as previously described (Funghetto et al., 2013;
© 2014 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd
Eccentric resistance exercise and obese elderly women, D. da C. Nascimento et al. 3
Tajra et al., 2013). Upon arrival to the lab, subjects began with a warm-up on a cycle ergometer for 10 min at 60 rpm and 50 W, followed by 10 leg extension repetitions at 50% of 10 RM load, and then a rest interval of 3–5 min was allowed. The ERE session was performed on a bilateral knee extension isoinertial machine with a load corresponding to 110% of the 10 RM. Subjects performed only the eccentric phase of the lift (2–3 s); at the end of each eccentric repetition, the researcher moved the load through the concentric portion of the movement to begin the next eccentric repetition. Subjects completed seven sets of 10 repetitions with a passive rest of 3 min between sets. The 10 RM trials and ERE session were scheduled between 2:00 and 4:00 p.m. and were performed under standardized controlled room temperature. Gelatin zymography Blood samples were drawn from the antecubital vein in the dominant arm by venipuncture at baseline, 0, 3, 24 and 48 h post ERE and collected in lithium heparin-coated vacutainer tubes without EDTA. Blood plasma was separated and immediately frozen and stored at 20°C in small volume aliquots until used for the assays described below. All blood samples, baseline and postexercise, were collected, handled, and stored (a)
under identical conditions. Plasma levels of pro-MMP-9 and pro-MMP-2 were further analysed through gelatin zymography (Dwek et al., 2012). All the subjects during zymography were together analysed at the same time point as shown in Fig. 1. The activity of MMP-2 and MMP-9 was determined by densitometric scanning of the bands (ImageScanner III, LabScan 6.0). The results are presented as band intensity, area and intensity versus area for MMP-2 and MMP-9. Interleukin-6 IL-6 concentration was measured by Quantikine or Quantikine high sensitivity commercial enzyme-linked immunosorbent assay Kit (R&D Systems, Minneapolis, MN, USA). The intraassay coefficient of variation of the kits was 15–56%, and the interassay coefficient of variation was 43–64%. The measures for IL-6 were performed in triplicate and average was used. Statistical analyses Normality of the data was determined by the Shapiro–Wilk normality test and the homoscedasticity test (Mauchly test). All variables presented normal distribution and homocedasticity. The repeated-measure ANOVA was used, and when differ(b)
(c)
Figure 1 Matrix metalloproteinase-2 (MMP-2) response to eccentric resistance exercise. Values presented by mean and standard deviation (SD), *significant different versus 3 h (P
