Background: Obesity is one of the most important health problems worldwide, several factors related to lifestyle as physical inactivity and unbalanced diets increase their development. This condition is characterized by low-chronic inflammation by excess of adipose tissue. The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) protein is part of NLRP3 inflammasome, a complex related to inflammation and metabolic alterations. Purpose: The aim of this study was to evaluate the effect of physical exercise program on ASC gene expression and inflammatory markers in obese adults. Methods: 37 obese individuals were randomized to exercise-diet group or diet-group during a 4-month follow-up period. The dietary evaluation was analyzed by Nutritionist Pro software. Body composition was evaluated by bioimpedance (InBody 370). All biochemical determinations were analyzed by dry chemistry (Vitros 350). ASC messenger ribonucleic acid (mRNA) expression was performed by real-time polymerase chain reaction (PCR) using Taqman probes and by the 2-ΔΔcq quantification method. Cytokine levels was performed using the Bio-PlexPro™ HumanTh17Cytokine Assays (MagPix) panel. Statistical analysis was performed with Statistical Package for the Social Sciences (SPSS) v.22 software.
A randomized clinical trial was conducted in the Institute of Translational Nutrigenetics and Nutrigenomics, Department of Molecular Biology in Medicine, Health Sciences Center, at the University of Guadalajara, Guadalajara, Jalisco, Mexico, from February of 2018 to February of 2019. All participants were recruited through flyers and social media invitations. Sample size was determined according to the mean difference formula for clinical trials. To achieve a statistical power of 80% and an alpha of 5%, a sample size of 13 participants in each study group was required. However, 37 obese individuals who met the selection criteria were randomized in exercise-diet group or diet-group. Blood sample, height and weight were measured after 8-12 hour fast and wearing light clothes. The participants in the exercise-diet group were cited in the Respiratory Unit of the University of Guadalajara with the indication of not consuming caffeine, tobacco or energy drinks 24 hours before the test, and not having eaten food at least two hours before the Astrand-rhyming Test. This study was approved by the Ethics and Biosafety Committee of the Health Sciences Center, University of Guadalajara (registry number CUCS/CINV/0476/18). The procedures were in accordance with this institution's guidelines and all the participants signed a written consent-informed.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
TREATMENT
Masking
NONE
Enrollment
37
The participants who performed exercise-diet intervention performed an Astrand-rhyming test before and after the exercise program intervention. Participants followed a progressive exercise-program during four months according to initial physical fitness. The main exercises consisted of improving aerobic, speed and resistance performance. During all single sessions, breathlessness and fatigue were measured using 0 to10 Borg scale. A personal trainer certified by American College of Sports Medicine (ACSM) supervised three sessions per week, the other two unsupervised training days the subjects were given the detailed training program to do in house/park. And also, this group received a diet intervention that consisted of a 20 percentage reduction of total energy expenditure estimated by the Mifflin formula with a nutrients distribution as follows: 50% carbohydrates, 30% lipids, and 20% proteins.
This group received a diet intervention that consisted of a 20 percentage reduction of total energy expenditure estimated by the Mifflin formula with a nutrients distribution as follows: 50% carbohydrates, 30% lipids, and 20% proteins.
Erika Martínez-López
Guadalajara, Jalisco, Mexico
Change in ASC mRNA Expression
The polymorphonuclear cells were separated using Dextran treatment from peripheral blood samples. Subsequently, TRIzol® reagent was using according to the manufacturer´s instructions. The complementary DNA (cDNA) synthesis was performed according to standard techniques. Quantitative real-time PCR was performed using TaqMan® probes in Light Cycler 96 equipment considering standards PCR conditions to analyze ASC mRNA relative expression by 2-ΔΔcq method. The amplification reactions were performed in duplicate using β-Actin gene as constitutive gene to normalized the samples.
Time frame: Mean change from baseline (0 Month) to end of treatment at 4th Month.
Change in Pro-inflammatory and Anti-inflammatory Cytokines
The pro-inflammatory and anti-inflammatory cytokines quantification were using Bio-Plex Pro™ Human cytokine Standard 17-Plex, Group I kit following the supplier's instructions, and the read was immediately by MAGPIX™ analyzer.
Time frame: Mean change from baseline (0 Month) to end of treatment at 4th Month.
Change in Astrand-rhyming Submaximal Test
The Astrand-rhyming submaximal test was performed as described by Astrand.
Time frame: Mean change from baseline (0 Month) to end of treatment at 4th Month.
Changes in Height
Height in meters using a stadiometer.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month. 1st month, 2nd month, 3th month and the 4th month.
Changes in Weight
The weight was measured in kilograms on InBody 370.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Body Mass Index (BMI)
Weight and height were be combined to report BMI in kg/m\^2
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Waist Circumference
Waist circumference was measured at the narrowest point between the edge of the inner rib and the iliac crest, with the participant in an abducted and relaxed position, after expiration using a Lufkin Executive® tape.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Fat Mass
The Fat Mass was measured in kilograms by electrical bioimpedance on InBody 370.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Musculoskeletal Mass
The musculoskeletal mass was measured in kilograms by electrical bioimpedance on InBody 370.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Serum Glucose
It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Serum Insulin
Were determined through Insulin Model ELISA kit following the supplier's instructions.
Time frame: At the baseline (0 Month), 3th month and the 4th month.
Changes in homeostatic model assessment - insulin resistance (HOMA-IR)
Serum glucose and Insulin levels were be combined to report HOMA-IR calculated as described by Matthews.
Time frame: At the baseline (0 Month), 3th month and the 4th month.
Changes in Total Cholesterol
It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in High-density lipoprotein (c-HDL)
It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment.
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
Changes in Atherogenic Index
Serum total cholesterol and c-HDL were be combined to report atherogenic index, calculated through formula \[Total cholesterol (mg/dL) / HDL-c (mg/dL)\].
Time frame: At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month.
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