
AOX: Antioxidant; BP: Blood Pressure; CVD: Cardiovascular disease; DBP: Diastolic Blood Pressure; ELISA: Enzyme-Linked Immune Sorbent Assay; GC: Glucocorticoids LDL: Low Density Lipoprotein; MDA: Malonaldehyde PJ: Pomegranate (pomegranate pure) Juice ROS: Reactive Oxygen Species SBP: Systolic Blood Pressure TBARS: Thio barbituric Reactive Substances
Free-living male and female volunteers (n = 24) who were apparently healthy participated in this study. Subjects were recruited from the University following an advertisement through the internal moderator email system. All participants had to meet the following criteria before enrolment in the study: no participation in regular physical activity (vigorous exercise two times or more per week); no chronic health problems or current smoking; no history of cardiovascular, metabolic, or respiratory disease and no consumption of antioxidant supplements within the past 6 months. Written informed consent was provided by each participant consistent with the university policy on the protection of human subjects. The protocol of the study was approved by the Research Ethics Committee at Queen Margaret University (UK).
Glucocorticoids
Twenty-four-hour urine samples were collected from volunteers into sterile plastic containers after each exercise session at baseline and after 1 week of the juice or placebo intake. The weight of each collected sample of urine was measured and a 20 mL aliquot was taken and stored at -80°C until analysis.
The TBARS assay was performed for the assessment of oxidative stress in urine as previously described [50]. This assay quantifies the amount of malonaldehyde (MDA) formed as a result of lipid peroxidation and involves reacting samples with 2-thiobarbituric acid (TBA) under high temperatures (90-100°C) and acidic conditions. TBA reacts with a MDA to produce a stable adduct that can be quantified spectrophotometrically.
A validated automated A&D Medical UA-767 BP monitor (A&D medical, San Jose, CA, USA) was used to measure BP according to Grassi., et al. [51]. Three readings of blood pressure were taken before and after each session and the average was calculated.
A record of all food and beverage intake was monitored over the study period to assess compliance with the dietary instructions given and also to determine whether nutritional intake was different between groups. Three-day dietary records were provided to each participant with standard instructions on how to complete the record. Participants were instructed to estimate servings of foods using household measurements as described in national dietary guidance documents as previously described [52]. Diet records were assessed using Windiets™ (2011) for windows (Robert Gordon University, Aberdeen) [53] and were analysed for macronutrient, micronutrient and caloric intake.
Placebo Group (n = 12) | Placebo Group (n = 12) | |||
Mean ± SD | Range | Mean ± SD | Range | |
Age (years) |
22.1 ± 4.6 | 20 - 34 | 22.8 ± 5.1 | 18 - 37 |
Weight (kg) |
79.7 ± 12.8 | 62.7 - 105.2 | 70.6 ± 10.5 | 55.8 - 90.3 |
Waist circumference (cm) |
89.6 ± 13.1 | 71 - 117 | 82.7 ± 11.6 |
68 - 110 |
Body mass index (kg/m2) |
26.9 ± 3.8 | 20.6 - 29.5 | 25.3 ± 3.7 | 20.1 - 29.4 |

Treatment Group (n = 12) | Placebo Group (n = 12) | ||||
Pre-exercise | Post-exercise | Pre-exercise | Post-exercise | ||
SBP Post Intervention |
Baseline | 136.7 ± 11.5 131.8 ± 8.8 * |
158.8 ± 15.8 148.1 ± 12.3** |
130.1 ± 14.1 131.7 ± 13.2 |
144.5 ± 11.7 145.1 ± 19.9 |
DBP Post Intervention |
baseline | 86.3 ± 8.1 82.5 ± 6.8+ |
103.1 ± 12.5 93.9 ± 11.5 |
85 ± 11.8 84.4 ± 9.4 |
92.1 ± 12.6 93.3 ± 10.5 |
Statistical significance within each group based on paired t-test: *P = 0.01, **P < 0.01, ‡P = 0.001, + P=0.04. Results are expressed as mean values ± SD, where n = 12.
Treatment Group (n = 12) | Placebo Group (n = 12) | |||
Parameter | Baseline | After 1-week | Baseline | After 1-week |
TBARS (µmole/day) | 31.2 ± 10.6 | 26.5 ± 9.8* | 27.1 ± 8.4 | 28.7 ± 10.7 |
Free Cortisol (nmole/day) | 179.4 ± 53.2 | 125.6 ± 43.5* | 166.6 ± 71.5 | 191.2 ± 93.7 |
Free Cortisone (nmole/day) | 112.2 ± 40.4 | 187.6 ± 90.2* | 125.5 ± 49.5 | 136.4 ± 53.8 |
Ratio: Cortisol/cortisone | 1.598 ± 1.1 | 0.669 ± 0.55** | 1.33 ± 0.44 | 1.41 ± 0.48 |
Statistical significance within each group based on paired t-test: * P < 0.05 and **P < 0-01. Results are expressed as mean values ± standard deviation (SD), where n = 12.
∆ SBP- Basal | ∆ DBP- Basal | ∆ SBP Post | ∆ DBP Post | ||||
Ratio Cortisol/cortisone | Correlation Coefficient | -0.347 | -0.231 | -0.490* | -0.573** | ||
Sig. (2-tailed) | 0.133 | 0.326 | 0.028 | 0.008 | |||
N | 12 | 12 | 12 | 12 |
** Correlation is significant at the 0.01 level (2-tailed). * Correlation is significant at the 0.05 level (2-tailed).
There was no significant change in either BMI or WC in both study groups after the 7 days’ intervention period. (BMI- Intervention: 26.72kg/m2 - 26.85 kg/m2 p = 0.278, Control: 25.67kg/m² - 25.61 kg/m² p = 0.509.) (WC- Intervention: 89.8 cm - 89.5 cm p = 0.810, Control: 82.6 cm - 82.7 cm). The analysis of the food diary during the phases of our study showed that there was a comparable intake of total energy and macronutrients in both the PJ and placebo groups suggesting that their food intake did not influence the physiological and biochemical parameters studied.

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