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Effects of Punica granatum L. peel extract supplementation on body weight, cardiac function, and haematological and biochemical parameters in an animal model of metabolic syndrome Cover

Effects of Punica granatum L. peel extract supplementation on body weight, cardiac function, and haematological and biochemical parameters in an animal model of metabolic syndrome

Journal of Veterinary Research
Volume 67 (2023): Issue 2 (June 2023)
By: Joanna Niewiadomska,  Ewa Kumiega,  Michał Płóciennik,  Jacek Gajek and  Agnieszka Noszczyk-Nowak  
Open Access
|Jun 2023

Figures & Tables

Fig. 1.

Violin plots of body weight with regard to individual groups at the final 16th-time point. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means
Violin plots of body weight with regard to individual groups at the final 16th-time point. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means

Fig. 2

The dynamics of body weight increases of individual rats (left side) and of group mean weight increases (right side) during the study period. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 - experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP
The dynamics of body weight increases of individual rats (left side) and of group mean weight increases (right side) during the study period. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 - experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP

Fig. 3

Trajectories of individual rats’ body weight variation and group average variations. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP
Trajectories of individual rats’ body weight variation and group average variations. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP

Fig. 4

Violin plot of the level of acanthocytes by group after eight weeks of pomegranate peel supplementation. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means
Violin plot of the level of acanthocytes by group after eight weeks of pomegranate peel supplementation. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means

Fig. 5

Violin plots of the level of schistocytes by group after eight weeks of pomegranate peel supplementation. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means
Violin plots of the level of schistocytes by group after eight weeks of pomegranate peel supplementation. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means

Fig. 6.

A – Acanthocytes and B – schistocytes found in smears of rat peripheral blood. Abnormal cells are indicated by arrows
A – Acanthocytes and B – schistocytes found in smears of rat peripheral blood. Abnormal cells are indicated by arrows

Fig. 7

Violin plots of fasting blood glucose concentrations by group after eight weeks of pomegranate peel supplementation. fa/fa H2O - control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 - experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 - experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 - experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 - experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means
Violin plots of fasting blood glucose concentrations by group after eight weeks of pomegranate peel supplementation. fa/fa H2O - control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 - experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 - experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 - experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 - experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means

Fig. 8.

Violin plots of lipid profiles by group after eight weeks of pomegranate peel supplementation. A – serum triglyceride concentration; B – serum total cholesterol concentration; C – serum high-density lipoprotein concentration; D – serum low-density lipoprotein concentration. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means
Violin plots of lipid profiles by group after eight weeks of pomegranate peel supplementation. A – serum triglyceride concentration; B – serum total cholesterol concentration; C – serum high-density lipoprotein concentration; D – serum low-density lipoprotein concentration. fa/fa H2O – control group of Zucker diabetic fatty (ZDF) rats with missense mutation in the Lepr leptin receptor gene administered only water; fa/fa 100 – experimental group of ZDF rats with this mutation administered 100 mg/kg body weight (b.w.) pomegranate peel extract (EPP); fa/fa 200 – experimental group of ZDF rats with this mutation administered 200 mg/kg b.w. EPP; fa/+ 100 – experimental group of ZDF rats without this mutation administered 100 mg/kg b.w. EPP; fa/+ 200 – experimental group of ZDF rats without this mutation administered 200 mg/kg b.w. EPP; μ^$${\hat \mu }$$ – predicted value of group means

Mean body weights for individual groups at the final 16th time point

groupfa/fa H2Ofa/fa 100fa/fa 200fa/+ 100fa/+ 200
n66666
Mean407.14382.08392.75358.97337.69
Standard deviation32.6835.4836.2812.9029.34

Glucose and lipid profile at the beginning of the study (t1) and after eight weeks of pomegranate supplementation (t2)

Indexfa/fa H2Ofa/fa 100fa/fa 200fa/+ 100fa/+ 200
t1t2t1t2t1t2t1t2t1t2
GLU (mmol/L)17.50 ± 5.9059.40 ± 11.3021.40 ± 6.4063.00 ± 12.8021.10 ± 8.5057.80 ± 8.507.60 ± 0.6023.00 ± 3.707.70 ± 1.2022.90 ± 2.80
CHOL (mmol/L)4.30 ± 0.305.10 ± 0.704.30 ± 0.204.80 ± 0.504.10 ± 0.105.10 ± 1.003.40 ± 0.202.20 ± 0.303.60 ± 0.102.40 ± 0.10
LDL (mmol/L)2.50 ± 0.102.90 ± 0.402.40 ± 0.203.00 ± 0.402.40 ± 0.203.20 ± 0.601.70 ± 0.101.30 ± 0.201.80 ± 0.101.40 ± 0.10
HDL (mmol/L)2.20 ± 0.102.80 ± 0.201.90 ± 0.102.80 ± 0.201.90 ± 0.202.80 ± 0.401.20 ± 0.201.20 ± 0.201.20 ± 0.101.10 ± 0.10
TG (mmol/L)11.50 ± 2.705.60 ± 5.6012.70 ± 3.405.70 ± 2.9012.50 ± 2.805.90 ± 1.301.30 ± 0.301.20 ± 0.601.30 ± 0.300.90 ± 0.60

Blood cell counts at two time points: at the beginning of the study (t1) and after eight weeks of EPP supplementation (t2)

Indexfa/fa H2Ofa/fa 100fa/fa 200fa/+ 100fa/+ 200
t1t2t1t2t1t2t1t2t1t2
WBC (K/μL)10.40 ± 1.637.05 ± 1.9610.50 ± 1.266.02 ± 2.3311.26 ± 2.566.40 ± 1.298.52 ± 0.865.00 ± 1.419.38 ± 0.993.73 ± 1.25
RBC (G/μL)7.49 ± 0.218.36 ± 0.697.63 ± 0.238.59 ± 0.447.60 ± 0.138.19 ± 0.537.53 ± 0.648.07 ± 0.547.69 ± 0.247.82 ± 0.63
HGB (g/L)16.13 ± 0.3714.53 ± 0.9916.10 ± 0.3714.80 ± 0.6515.90 ± 0.3414.32 ± 0.7315.70 ± 0.7313.88 ± 0.8816.05 ± 0.4213.90 ± 0.35
HCT (%)43.47 ± 1.2543.23 ± 3.3139.18 ± 12.4944.52 ± 2.3244.12 ± 0.9242.22 ± 2.6641.88 ± 3.7641.86 ± 2.6142.75 ± 1.4839.75 ± 3.18
PLT (K/μL)1,069.33 ± 207.16748.00 ± 76.581033.83 ± 74.09658.67 ± 67.821078.80 ± 76.59718.20 ± 52.11905.80 ± 53.27656.60 ± 244.39911.67 ± 60.07738.00 ± 39.33
MCV (fL)58.00 ± 1.1051.83 ± 0.7557.67 ± 0.5251.83 ± 0.4158.00 ± 1.0051.60 ± 0.5555.60 ± 1.1451.80 ± 0.8455.50 ± 0.5551.00 ± 0.00
MCH (pg)21.55 ± 0.5817.43 ± 0.6321.15 ± 0.2217.25 ± 0.2920.88 ± 0.4017.50 ± 0.4220.90 ± 1.0717.18 ± 0.2620.90 ± 0.1917.87 ± 1.11
MCHC (g/dL)37.15 ± 0.6933.65 ± 1.1336.50 ± 0.4933.30 ± 0.6236.02 ± 0.2933.92 ± 0.7337.62 ± 2.0033.20 ± 0.1737.58 ± 0.4235.08 ± 2.23
RET (%)2.17 ± 0.871.85 ± 1.562.72 ± 0.311.15 ± 0.823.60 ± 0.670.92 ± 0.332.34 ± 0.611.12 ± 1.111.48 ± 0.690.53 ± 0.30
NEU (%)13.33 ± 6.9225.50 ± 4.7619.33 ± 8.0421.50 ± 4.5920.40 ± 5.0833.20 ± 5.7216.25 ± 7.3720.75 ± 3.9512.83 ± 5.5318.67 ± 4.50
LYM (%)81.17 ± 5.0071.50 ± 5.6876.83 ± 10.2674.83 ± 6.1178.40 ± 5.1361.80 ± 4.7681.00 ± 8.6875.75 ± 5.6884.67 ± 4.6379.50 ± 4.46
MONO (%)2.50 ± 3.510.83 ± 0.981.00 ± 0.890.33 ± 0.520.20 ± 0.452.00 ± 1.580.50 ± 1.000.25 ± 0.501.83 ± 1.330.17 ± 0.41
EOS (%)1.83 ± 0.750.67 ± 0.521.50 ± 1.382.33 ± 2.420.60 ± 0.551.60 ± 1.342.00 ± 1.152.50 ± 1.290.33 ± 0.521.33 ± 1.03

Mass spectrum characteristic and content of phenolic compounds in pomegranate peel extract

Rt (min)MS (M-H)− (m/z)MS/MS (M-H)− (m/z)Name of compoundsPolyphenol content
1.67331271/169Galloyl-glucose2.00 ± 0.03
1.73781721/601Punicalin α/A3.11 ± 0.06
2.021083611/331/146HHDP-galloyl-hexoside (punicalagin)4.20 ± 0.09
2.121083781/622/301Punicalagin isomer14.82 ± 1.04
2.33933631/450/301Ellagitannin4.71 ± 0.40
2.871083781/301HHDP-gallagyl-hexoside (punicalagin)93.91 ± 2.05
3.121085907/783/301Ellagic acid derivative2.49 ± 0.53
3.691083781/301HHDP-gallagyl-hexoside (punicalagin)157.00 ± 2.65
3.89799301Granatin A4.74 ± 0.32
5.08783481/301Ellagitannin25.86 ± 1.53
6.201085933/301Digalloyl-gallagyl-hexoside10.37 ± 0.65
6.25783481/301Ellagitannin13.51 ± 0.99
6.38463301Ellagic acid-hexoside33.63 ± 1.23
6.89951907/635/301Galloyl-HHDP-DHHDP-hex (granatin B)2.68 ± 0.11
Total (mg/g dry weight)373.05

Echocardiographic parameters at two time points: at the beginning of the study (t1) and after eight weeks of pomegranate peel supplementation (t2)

Indexfa/fa H2Ofa/fa 100fa/fa 200fa/+ 100fa/+ 200
t1t2t1t2t1t2t1t2t1t2
HR (bpm)289.00 ± 28.08234.33 ± 37.03265.00 ± 21.02232.25 ± 45.85245.17 ± 21.34226.83 ± 32.62264.60 ± 20.40230.60 ± 46.55275.20 ± 11.82245.80 ± 31.40
LVIDs (mm)2.52 ± 0.542.65 ± 1.033.35 ± 1.612.65 ± 1.033.35 ± 1.232.90 ± 1.013.36 ± 0.692.36 ± 1.192.80 ± 0.702.28 ± 0.42
LVIDd (mm)6.48 ± 0.516.08 ± 1.156.65 ± 0.835.82 ± 0.746.87 ± 1.256.52 ± 0.956.08 ± 1.135.56 ± 1.395.84 ± 0.775.68 ± 0.69
LVPWs (mm)2.93 ± 0.323.00 ± 0.392.72 ± 0.433.00 ± 0.552.62 ± 0.392.77 ± 0.672.68 ± 0.493.32 ± 0.722.38 ± 0.493.18 ± 0.72
LVPWd (mm)1.80 ± 0.391.95 ± 0.421.78 ± 0.392.22 ± 0.381.88 ± 0.282.03 ± 0.431.98 ± 0.561.86 ± 0.191.62 ± 0.301.94 ± 0.42
IVSs (mm)3.32 ± 0.383.15 ± 0.623.10 ± 0.553.67 ± 0.713.68 ± 0.553.22 ± 0.413.04 ± 1.103.24 ± 0.583.30 ± 0.713.38 ± 0.36
IVSd (mm)1.77 ± 0.161.77 ± 0.291.73 ± 0.132.00 ± 0.271.92 ± 0.231.80 ± 0.431.96 ± 0.551.62 ± 0.461.78 ± 0.191.86 ± 0.30
LVEDV (mL)0.63 ± 0.150.55 ± 0.260.68 ± 0.220.48 ± 0.220.80 ± 0.380.65 ± 0.260.54 ± 0.260.44 ± 0.370.50 ± 0.160.44 ± 0.15
LVESV (mL)0.05 ± 0.050.07 ± 0.050.15 ± 0.170.05 ± 0.100.13 ± 0.120.08 ± 0.080.10 ± 0.070.04 ± 0.090.06 ± 0.090.02 ± 0.04
SV (mL)0.60 ± 0.140.50 ± 0.240.58 ± 0.100.40 ± 0.220.65 ± 0.310.57 ± 0.200.44 ± 0.210.40 ± 0.280.40 ± 0.160.38 ± 0.13
CO (L/min)0.17 ± 0.040.12 ± 0.070.15 ± 0.020.09 ± 0.040.16 ± 0.070.14 ± 0.060.12 ± 0.060.09 ± 0.040.11 ± 0.040.10 ± 0.03
EF (%)92.85 ± 3.3689.83 ± 6.3883.50 ± 14.0184.12 ± 18.6485.80 ± 8.9289.55 ± 5.7180.68 ± 7.1890.22 ± 6.4087.00 ± 7.2692.64 ± 1.78
FS (%)61.25 ± 6.7857.72 ± 12.3851.52 ± 18.5553.75 ± 21.0952.07 ± 12.0956.62 ± 10.4744.80 ± 8.1258.58 ± 13.6852.18 ± 8.2959.94 ± 3.22
mFS (%)25.40 ± 3.6722.05 ± 3.2421.50 ± 7.1418.10 ± 12.3120.95 ± 4.6725.25 ± 3.2518.84 ± 3.4315.22 ± 3.3719.92 ± 1.7619.80 ± 7.22
LAD (mm)4.22 ± 0.713.57 ± 0.474.34 ± 1.083.82 ± 0.364.87 ± 0.533.83 ± 0.694.30 ± 0.584.15 ± 0.424.58 ± 0.423.76 ± 0.29
AOD (mm)3.37 ± 0.383.23 ± 0.353.34 ± 0.733.44 ± 0.253.85 ± 0.233.12 ± 0.513.28 ± 0.393.65 ± 0.833.44 ± 0.223.12 ± 0.33
LA/Ao1.25 ± 0.191.10 ± 0.081.30 ± 0.111.12 ± 0.131.26 ± 0.111.24 ± 0.171.31 ± 0.081.17 ± 0.181.34 ± 0.181.21 ± 0.13
DOI: https://doi.org/10.2478/jvetres-2023-0031 | Journal eISSN: 2450-8608
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Language: English
Page range: 219 - 232
Submitted on: Jan 13, 2023
Accepted on: May 15, 2023
Published on: Jun 16, 2023
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
polyphenols,
pomegranate peel,
Zucker diabetic fatty rats,
metabolic syndrome
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2023 Joanna Niewiadomska, Ewa Kumiega, Michał Płóciennik, Jacek Gajek, Agnieszka Noszczyk-Nowak, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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