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

Karabulut, Gözde; Barlas, Nurhayat

doi:10.2478/aiht-2022-73-3617

Figures & Tables

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

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

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)
Parameters	Control	25	50	100	200	400
ALT (IU/L)	45.65±17.41	50.3±0.22	55.4±0.11	65.25±15.99^a,b	85.63±10.72^a,b,c	96.91±7.82^a,b,c,d
AST (IU/L)	110.1±101.6	115.2±15.12	120.45±22.12^d,e,f	139.8±88.9^a	182.6±36.9^a,b	268.3±110.6^a,b,c
Triglycerides (mg/dL)	945.8±704.6	1050.27±5.43	1210.12±3.54^a,d	1450.8±294.5^a	1650.3±250^a,b	1702.6±266.4^a,b,c
Glucose (IU/L)	191.5±61	201.27±13.07	205.17±0.6	211.1±33.5^a,e,f	251.1±19.3^a,b	301.6±43.5^a,b,c
Total protein (g/dL)	8.6±0.8	8.3±2.34	8.22±3.34	8.5±2.6	6.8±0.3^a,b,c,d	6.6±0.4^a,b,c,d
Albumin (g/dL)	32.35±2.73	29.8±4.44	29.8±1.14	27.93±1.34	21.03±1.88^a,b,c,d	18.66±1.53^a,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±2^a,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)
Groups	Control	25	50	100	200	400
Insulin (ng/mL)	1.43±0.48	1.35±0.12	1.32±0.1	1.24±1.03^a	0.77± 0.82^a,b,c,d	0.69±0.61^a,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)
Groups	Control	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±6.51^a,b,c,f	30.05±5.72^a,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)
Groups	Control	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±0.9^a,b	17.9±0.1^a,b	18.44±0.1^a,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±0.1^a,b
Relative weight (mg/g)	0.74±0.1	0.73±0.1	0.73±0.1	0.71±0.2	0.68±0.3^a,b,c,d	0.66±0.5^a,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)
Groups	Control	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±0.9^a	16.9±0.1^a	17.48±0.1^a
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±0.3^a,b,c	0.160±0.4^a,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±0.3^a,b,c,d	0.59±0.5^a,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)
Parameters	Control	25	50	100	200	400
ALT (IU/L)	45.65±17.41	47.26±0.22	50.62±0.11	55.25±14.99^a	76.63±19.72^a,b,c	82.91±6.82^a,b,c,d
AST (IU/L)	110.1±101.6	120.9±15.12	135.66±22.12	159.8±65.9^a	191.6±36.9^a,b	276.3±90.6^a,b,c,d
Triglycerides (mg/dL)	945.8±704.6	960.8±5.43	1100.9±3.54	1550.8±294^a,b	1850.3±250^a,b,c	1820.4±116.4^a,b,c
Glucose (IU/L)	191.5±61	220.27±13.07	251.17±0.6	291.1±22.5^a	351.1±18.3^a,b	401.6±41.5^a,b,c
Total Protein (g/dL)	8.6± 0.8	8.3±2.34	8.2±3.34	8.1±2.6	7.1±0.3^ab	6.4±0.4^a,b,c
Albumin (g/dL)	32.35±2.73	31.9±4.44	31.2±1.14	29.92±1.32	22.03±1.98^a,b,c	21.33±1.43^a,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±0.14^a,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±2^a,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)
Parameters / Groups	Control	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)
Parameters / Groups	Control	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)
Groups	Control	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±6.51^a	23.10±5.72^a,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)
Parameters	Control	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±0.17^a,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±0.11^a,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±2.89^a,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±0.22^a,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)
Groups	Control	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	±0.71^a,b,c,d	0.80±0.51^a,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)
Parameters	Control	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±1.89^a,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±0.93^a,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

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

Figures & Tables

Figure 1

Figure 2

Figure 3

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

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

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

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

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

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

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

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

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

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

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

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

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

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