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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: ,  ,  ,   and    
Open Access
|Jun 2023
Figures & tables

Figures & Tables

Table 1.

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

1 Rt – retention time; MS – mass spectrometry; (M-H)− – deprotonated molecule; m/z – mass-to-charge ratio; MS/MS – tandem mass spectrometry; HHDP – hexahydroxydiphenic acid; DHHDP – dehydrohexahydroxydiphenic acid

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^ – predicted value of group means

Table 2.

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

1 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. 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

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

Table 3.

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

1 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; HR – heart rate; LVIDs – left ventricular internal dimension at end systole; LVIDd – left ventricular internal dimension at end diastole; LVPWs – left ventricular posterior wall thickness at end systole; LVPWd – left ventricular posterior wall thickness at end diastole; IVSs – interventricular septum thickness at end systole; IVSd – interventricular septum thickness at end diastole; LVEDV – left ventricular end-diastolic volume; LVESV – left ventricular end-systolic volume; SV – stroke volume; CO – cardiac output; EF - ejection fraction; FS – fractional shortening; mFS – mid-wall fractional shortening; LAD – left atrial diameter; AOD – aortic diameter; LA/Ao – left atrial to aortic root ratio. Values are presented as mean ± standard deviation

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^ – 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^ – 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

Table 4.

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

1 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; WBC – white blood cell count; RBC – red blood cell count; HGB – haemoglobin concentration; HCT – hematocrit; PLT – platelet count; MCV – mean corpuscular volume; MCH – mean corpuscular haemoglobin; MCHC – mean corpuscular hemoglobin concentration; RET – percent of reticulocytes; NEU –- percent of neutrophils; LYM – percent of lymphocytes; MONO – percent of monocytes; EOS - percent of eosinophils. Values are presented as mean ± standard deviation

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^ – predicted value of group means

graphic/j_jvetres-2023-0031_fig_008.jpg
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^ – predicted value of group means

Table 5.

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

1 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; GLU - glucose concentration; CHOL - total cholesterol concentration; LDL – low-density lipoprotein concentration; HDL – high-density lipoprotein concentration; TG - triglyceride concentration. Values are presented as mean ± standard deviation

DOI: https://doi.org/10.2478/jvetres-2023-0031 | Journal eISSN: 2450-8608 (formerly 2300-3235)
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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
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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