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Endocrine adverse effects of mono(2-ethylhexyl) phthalate and monobutyl phthalate in male pubertal rats

Gözde Karabulut
Gözde Karabulut
 and 
Nurhayat Barlas
Nurhayat Barlas
   | Dec 30, 2022
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Article Category: Original article
Published Online: Dec 30, 2022
Page range: 285 - 296
Received: Dec 01, 2021
Accepted: Nov 01, 2022
DOI: https://doi.org/10.2478/aiht-2022-73-3617
© 2022 Gözde Karabulut and Nurhayat Barlas, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Liver tissue photomicrographs in A) – the oil control group (×400 magnification); B and C – the group receiving MEHP dose of 100 mg/kg bw a day (×400 and ×200 magnification, respectively); D – the group receiving MEHP dose of 200 mg/kg bw a day (×200 magnification); E – the group receiving MEHP dose of 400 mg/kg bw a day (×200 magnification): (F) 200 mg/kg bw a day of monobutyl phthalate (MBP) (×400), (G) 400 mg/kg bw a day of MBP (×400). c – congestion; cd – cytoplasmic dissolution; mci – mononuclear cell infiltration; sd – sinusoidal degeneration
Liver tissue photomicrographs in A) – the oil control group (×400 magnification); B and C – the group receiving MEHP dose of 100 mg/kg bw a day (×400 and ×200 magnification, respectively); D – the group receiving MEHP dose of 200 mg/kg bw a day (×200 magnification); E – the group receiving MEHP dose of 400 mg/kg bw a day (×200 magnification): (F) 200 mg/kg bw a day of monobutyl phthalate (MBP) (×400), (G) 400 mg/kg bw a day of MBP (×400). c – congestion; cd – cytoplasmic dissolution; mci – mononuclear cell infiltration; sd – sinusoidal degeneration

Figure 2

Kidney tissue photomicrographs in A – the oil control group (×400 magnification); B – the group receiving MEHP dose of 100 mg/kg bw a day (×200 magnification); C – the group receiving MEHP dose of 200 mg/kg bw a day (×200 magnification); D – the group receiving MBP dose of 200 mg/kg bw a day (×400 magnification); E – the group receiving MBP dose of 400 mg/kg bw a day (×400 magnification). ag – athropic glomerulus; c – congestion; gd – glomerular degeneration; pvl – parietal and visceral leaves; td – tubular degeneration
Kidney tissue photomicrographs in A – the oil control group (×400 magnification); B – the group receiving MEHP dose of 100 mg/kg bw a day (×200 magnification); C – the group receiving MEHP dose of 200 mg/kg bw a day (×200 magnification); D – the group receiving MBP dose of 200 mg/kg bw a day (×400 magnification); E – the group receiving MBP dose of 400 mg/kg bw a day (×400 magnification). ag – athropic glomerulus; c – congestion; gd – glomerular degeneration; pvl – parietal and visceral leaves; td – tubular degeneration

Figure 3

Pancreas tissue photomicrographs in A – the oil control group (×400 magnification); B – the group receiving MEHP dose of 200 mg/kg bw a day (×400 magnification); C – the group receiving MEHP dose of 400 mg/kg bw a day (×400 magnification); D – the group receiving MBP dose of 200 mg/kg bw a day (×200 magnification); E – the group receiving MBP dose of 400 mg/kg bw a day (×200 magnification). c – congestion; dli – degeneration of Langerhans islets
Pancreas tissue photomicrographs in A – the oil control group (×400 magnification); B – the group receiving MEHP dose of 200 mg/kg bw a day (×400 magnification); C – the group receiving MEHP dose of 400 mg/kg bw a day (×400 magnification); D – the group receiving MBP dose of 200 mg/kg bw a day (×200 magnification); E – the group receiving MBP dose of 400 mg/kg bw a day (×200 magnification). c – congestion; dli – degeneration of Langerhans islets

Biochemical markers measured in the oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Parameters Control MEHP (mg/kg bw a day)
25 50 100 200 400
ALT (IU/L) 45.65±17.41 50.3±0.22 55.4±0.11 65.25±15.99a,b 85.63±10.72a,b,c 96.91±7.82a,b,c,d
AST (IU/L) 110.1±101.6 115.2±15.12 120.45±22.12d,e,f 139.8±88.9a 182.6±36.9a,b 268.3±110.6a,b,c
Triglycerides (mg/dL) 945.8±704.6 1050.27±5.43 1210.12±3.54a,d 1450.8±294.5a 1650.3±250a,b 1702.6±266.4a,b,c
Glucose (IU/L) 191.5±61 201.27±13.07 205.17±0.6 211.1±33.5a,e,f 251.1±19.3a,b 301.6±43.5a,b,c
Total protein (g/dL) 8.6±0.8 8.3±2.34 8.22±3.34 8.5±2.6 6.8±0.3a,b,c,d 6.6±0.4a,b,c,d
Albumin (g/dL) 32.35±2.73 29.8±4.44 29.8±1.14 27.93±1.34 21.03±1.88a,b,c,d 18.66±1.53a,b,c,d
Creatinine (mg/dL) 0.46±0.22 0.47±8.91 0.46±6.45 0.49±0.25 0.48±0.16 0.45±0.15
Urea (mg/dL) 14.8±3.3 14.7±4.12 13.09±2.44 13.7±1.8 12.3±8.3 11.5±2a,b,c

Insulin levels measured in oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Groups Control MEHP (mg/kg bw a day)
25 50 100 200 400
Insulin (ng/mL) 1.43±0.48 1.35±0.12 1.32±0.1 1.24±1.03a 0.77± 0.82a,b,c,d 0.69±0.61a,b,c,d

Food and water consumption by rats in the oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Groups Control MEHP (mg/kg bw a day)
25 50 100 200 400
Food (g) 10.70±3.60 11.12±1.01 12.14±2.29 15.43±4.37 20.06±6.51a,b,c,f 30.05±5.72a,b,c,d,e
Water (mL) 9.82±2.76 10.21±0.12 10.32±0.78 11.63±2.95 11.53±1.39 12.13±3.81

Body and absolute and relative organ weights of rats in the oil control and experimental groups treated with monobutyl phthalate (MBP)

Groups Control MBP (mg/kg bw a day)
25 50 100 200 400
Baseline body weight (g) 220.8±1.0 220.4±0.1 221.3±0.2 224.4±0.8 219.7±0.1 217.9±0.1
Final body weight (g) 330.9±1.2 331.02±0.8 332.01±0.3 323.2±0.1 330.4±0.4 350.7±0.1
Weight gain % 50.02±0.2 50.09±0.7 51.07±0.8 50.09±0.8 54.01±0.2 55.06±0.3
Liver
Absolute weight (g) 10.5±1.0 11.5±0.1 12.4±0.3 17.4±0.9a,b 17.9±0.1a,b 18.44±0.1a,b,c
Relative weight (mg/g) 34.8±1.2 33.3±1.3 33.1±1.2 32.8±0.7 33.9±0.4 34.1±1.1
Kidney
Absolute weight (g) 1.07±0.1 1.09±1.1 1.10±1.2 1.09±0.9 1.10±0.1 1.11±0.1
Relative weight (mg/g) 3.2±0.1 3.1±0.1 3.2±0.3 3.1±0.2 3.1±0.1 3.1±0.2
Pancreas
Absolute weight (g) 0.253±0.1 0.245±0.4 0.240±0.3 0.220±0.2 0.210±0.3 0.201±0.1a,b
Relative weight (mg/g) 0.74±0.1 0.73±0.1 0.73±0.1 0.71±0.2 0.68±0.3a,b,c,d 0.66±0.5a,b,c,d

Body and absolute and relative organ weights of rats in the oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Groups Control MEHP (mg/kg bw a day)
25 50 100 200 400
Baseline body weight (g) 220.8±1.0 219.5±0.2 221.1±0.1 222.3±0.9 217.8±0.1 218.9±0.1
Final body weight (g) 330.9±1.2 340.4±0.3 340.3±0.5 333.2±0.7 340.5±0.4 348.7±1.1
Weight gain % 50.02±0.2 52.03±0.5 51.02±0.1 51.09±0.8 55.01±0.3 54.06±0.4
Liver
Absolute weight (g) 10.5±1.0 12.4±1.2 14.2±0.1 16.5±0.9a 16.9±0.1a 17.48±0.1a
Relative weight (mg/g) 34.8±1.2 32.1±0.2 33.1±0.4 32.7±0.7 32.9±0.4 33.1±1.1
Kidney
Absolute weight (g) 1.07±0.1 1.07±0.1 1.09±0.2 1.09±0.9 1.09±0.2 1.10±0.1
Relative weight (mg/g) 3.2±0.1 3.1±0.2 3.2±0.2 3.1±0.2 3.1±0.1 3.0±0.2
Pancreas
Absolute weight (g) 0.253±0.1 0.240±0.2 0.242±0.4 0.212±0.2 0.198±0.3a,b,c 0.160±0.4a,b,c,d
Relative weight (mg/g) 0.74±0.1 0.73±0.2 0.72±0.1 0.71±0.2 0.68±0.3a,b,c,d 0.59±0.5a,b,c,d

Biochemical markers measured in the oil control and experimental groups treated with monobutyl phthalate (MBP)

Parameters Control MBP (mg/kg bw a day)
25 50 100 200 400
ALT (IU/L) 45.65±17.41 47.26±0.22 50.62±0.11 55.25±14.99a 76.63±19.72a,b,c 82.91±6.82a,b,c,d
AST (IU/L) 110.1±101.6 120.9±15.12 135.66±22.12 159.8±65.9a 191.6±36.9a,b 276.3±90.6a,b,c,d
Triglycerides (mg/dL) 945.8±704.6 960.8±5.43 1100.9±3.54 1550.8±294a,b 1850.3±250a,b,c 1820.4±116.4a,b,c
Glucose (IU/L) 191.5±61 220.27±13.07 251.17±0.6 291.1±22.5a 351.1±18.3a,b 401.6±41.5a,b,c
Total Protein (g/dL) 8.6± 0.8 8.3±2.34 8.2±3.34 8.1±2.6 7.1±0.3ab 6.4±0.4a,b,c
Albumin (g/dL) 32.35±2.73 31.9±4.44 31.2±1.14 29.92±1.32 22.03±1.98a,b,c 21.33±1.43a,b,c
Creatinine (mg/dL) 0.46±0.22 0.47±8.91 0.48±6.45 0.48±0.15 0.38±0.17 0.35±0.14a,b,c
Urea (mg/dL) 14.8±3.3 14.5±4.12 14.3±2.44 14.8±1.8 13.2±8.3 11.3±2a,b,d

Incidence of exposure-related histopathologic lesions observed in oil control and experimental groups treated with monobutyl phthalate (MBP)

Parameters / Groups Control MBP (mg/kg bw a day)
25 50 100 200 400
Liver
Sinusoidal degeneration 0/6 0/6 1/6 4/6* 4/6* 6/6*
Congestion 1/6 2/6 2/6 4/6* 3/6 5/6*
Cytoplasmic dissolution 0/6 1/6 1/6 3/6 3/6* 3/6
Mononuclear cell infiltration 1/6 0/6 2/6 2/6 5/6* 5/6*
Kidney
Glomerulus degeneration 0/6 0/6 1/6 3/6 4/6* 5/6*
Congestion 1/6 2/6 2/6 4/6* 3/6 6/6*
Cytoplasmic dissolution 0/6 1/6 1/6 1/6 1/6 2/6
Mononuclear cell infiltration 1/6 0/6 2/6 1/6 1/6 1/6
Pancreas
Degeneration of pancreatic cells 0/6 0/6 1/6 2/6 5/6* 6/6*
Congestion 1/6 1/6 1/6 2/6 2/6 5/6*

Incidence of exposure-related histopathological lesions observed in oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Parameters / Groups Control MEHP (mg/kg bw a day)
25 50 100 200 400
Liver
Sinusoidal degeneration 0/6 0/6 1/6 4/6* 5/6* 6/6*
Congestion 0/6 1/6 1/6 5/6* 3/6 5/6*
Cytoplasmic dissolution 0/6 1/6 1/6 1/6 4/6* 2/6
Mononuclear cell infiltration 0/6 0/6 1/6 4/6* 5/6* 5/6*
Kidney
Glomerulus degeneration 0/6 0/6 1/6 3/6 4/6 6/6*
Congestion 1/6 2/6 2/6 4/6* 3/6 5/6*
Cytoplasmic dissolution 0/6 1/6 1/6 3/6 3/6 3/6
Mononuclear cell infiltration 1/6 0/6 2/6 1/6 2/6 1/6
Pancreas
Degeneration of pancreatic cells 0/6 0/6 1/6 4/6* 4/6* 6/6*
Congestion 1/6 1/6 2/6 4/6* 3/6 5/6*

Food and water consumption by rats in the oil control and experimental groups treated with monobutyl phthalate (MBP)

Groups Control MBP (mg/kg bw a day)
25 50 100 200 400
Food (g) 10.70±3.60 12.18±1.03 13.67±2.32 14.23±4.37 18.07±6.51a 23.10±5.72a,b,c,d
Water (mL) 9.82±2.76 10.01±0.54 10.34±0.33 10.23±2.95 12.53±1.39 13.53±3.81

Haematological parameters measured in the oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP)

Parameters Control MEHP (mg/kg bw a day)
25 50 100 200 400
Leukocytes (mm3) 2.55±1.43 2.50±0.22 2.53±0.11 2.98±2.34 2.63±1.11 1.19±0.17a,b,c,d,e
Lymphocytes (%) 68.50±16.17 68.50±15.12 69.45±22.12 70.83±21.34 65.42±8.39 78.41±16.90
Monocytes (%) 11.17±5.40 10.27±5.43 9.12±3.54 11.40±8.42 10.30±1.97 4.40±0.11a,b,c,d,e
Neutrophils (%) 19.27±11.06 18.27±13.07 20.17±0.6 17.73±13.12 24.28±6.89 17.48±12.79
Erythrocytes (mm3) 13.22±3.53 14.23±2.34 14.22±3.34 11.6±2.68 13.3±1.76 8.26±0.68
MCV (fL) 27.8±5.34 29.8±4.44 28.8±1.14 29.41±1.13 25.78±3.89 53.12±2.89a,b,c,d,e
Haematocrit (%) 35.27±8.18 33.17±8.91 34.56±6.45 33.6±6.06 27.1±6.29 39.27±1.62
MCH (pg) 7.87±4.52 8.77±4.12 9.01±2.44 8.83±3.38 11.6±11.24 14.9±1.24
MCHC (g/dL) 26.75±8.78 25.15±11.22 28.65±7.55 29.8±9.16 23.05±6.05 28.07±1.06
Haemoglobin (g/dL) 8.92±1.26 8.92±0.1 8.44±1.77 8.50±1.16 8.05±0.5 6.33±0.22a,b,c,d,e
Thrombocytes (mm3) 3287.83±1276.73 3367.83±1131.02 3266.83±1144.03 3299.33±2483.02 2265±1567.18 2764.17±1876.21
PCT (%) 3.92±1.45 4.62±1.76 4.12±1.22 4.5±2.31 3.96±1.72 4.01±1.42

Antibody levels measured in oil control and experimental groups treated with mono(2-ethylhexyl) phthalate (MEHP) and monobutyl phthalate (MBP)

Groups Islet cell antibody (ICA) Insulin antibody (IAA)
Control (-) (-)
MEHP (mg/kg bw a day)
25 (-) (-)
50 (-) (-)
100 (-) (-)
200 (-) (-)
400 (-) (-)
MBP (mg/kg bw a day)
25 (-) (-)
50 (-) (-)
100 (-) (-)
200 (-) (-)
400 (-) (-)

Insulin levels measured in oil control and experimental groups treated with monobutyl phthalate (MBP)

Groups Control MBP (mg/kg bw a day)
25 50 100 200 400
Insulin (ng/mL) 1.43±0.48 1.40±0.70 1.38±0.60 1.33±1.01 0.81 ±0.71a,b,c,d 0.80±0.51a,b,c,d

Haematological parameters measured in the oil control and experimental groups treated with monobutyl phthalate (MBP)

Parameters Control MBP (mg/kg bw a day)
25 50 100 200 400
Leukocytes (mm3) 2.55±1.43 2.30±2.14 2.55±0.84 2.58±2.34 2.83±1.11 2.19±0.39
Lymphocytes (%) 68.50±16.17 67.51±17.17 70.60±26.01 71.83±21.34 72.42±8.39 73.41±16.90
Monocytes (%) 11.17±5.40 12.17±4.11 11.21±4.39 12.20±9.41 11.30±1.97 9.33±0.10
Neutrophils (%) 19.27±11.06 18.11±9.87 18.44±8.77 18.73±14.22 19.19±6.89 18.48±12.79
Erythrocytes (mm3) 13.22±3.53 12.12±3.21 13.01±1.21 12.6±2.68 13.3±1.76 12.26±0.68
MCV (fL) 27.8±5.34 28.9±4.53 28.8±1.01 28.42±2.13 29.78±3.89 62.11±1.89a,b,c,d,e
Haematocrit (%) 35.27±8.18 34.10±1.21 35.09±5.23 34.7±6.06 37.1±6.29 38.27±3.65
MCH (pg) 7.87±4.52 8.17±2.33 8.41±1.43 7.93±3.39 9.6±11.24 8.9±1.24
MCHC (g/dL) 26.75±8.78 27.15±5.56 27.31±2.56 27.2±9.16 25.05±6.05 26.07±1.06
Haemoglobin (g/dL) 8.92±1.26 9.02±0.26 9.98±0.26 9.5±1.16 9.02±1.6 6.01±0.93a,b,c,d,e
Thrombocytes (mm3) 3287.83±1276.7 3311.83±1222.01 3317.02±1143.3 3309.33±2483.0 3266±1598.1 3765.17±1659.2
PCT (%) 3.92±1.45 3.81±1.11 4.01±0.22 4.7±3.34 3.87±1.89 3.51±1.42
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