Fertaric acid amends bisphenol A-induced toxicity, DNA breakdown, and histopathological changes in the liver, kidney, and testis

Table 1 Protective effect of fertaric acid on body weight, food and water intake, urine volume, and urinary protein, albumin, and albumin/protein ratio in rats exposed to bisphenol A

Parameter	Group
	Control	Paraffin oil	FA	BPA	FA + BPA
Initial body weight (g)	132.5 ± 14.5	133.1 ± 15.6	131.8 ± 14.2	134.8 ± 13.9	135.0 ± 15.3
Final body weight (g)	185.0 ± 16.8	187.1 ± 17.2	186.3 ± 18.64	87.8 ± 12.5d	183.75 ± 16.7b
Initial food consumption (g/d)	11.5 ± 1.3	11.3 ± 1.0	11.4 ± 1.2	11.7 ± 1.1	11.6 ± 1.4
Final food consumption (g/d)	14.1 ± 1.1	14.3 ± 1.4	14.2 ± 1.3	5.7 ± 1.0d	14.0 ± 1.2b
Initial water intake (mL/d)	12.2 ± 1.5	12.4 ± 1.3	12.1 ± 1.2	12.3 ± 1.4	12.5 ± 1.0b
Final water intake (mL/d)	15.4 ± 1.6	15.2 ± 1.3	15.3 ± 1.0	6.8 ± 0.9d	15.1 ± 1.2b
Urine volume (mL/100 g/8 h)	0.98 ± 0.07	0.96 ± 0.09	0.99 ± 0.08	1.26 ± 0.15c	1.01 ± 0.08a
Urinary protein excretion (g/dL)	4.06 ± 0.24	4.04 ± 0.21	4.07 ± 0.26	5.29 ± 0.19c	4.08 ± 0.25a
Urinary albumin excretion (g/dL)	2.30 ± 1.4	2.28 ± 1.1	2.32 ± 1.3	4.13 ± 1.5d	2.31 ± 1.2b
Urinary albumin/protein excretion ratio	0.57 ± 0.05	0.56 ± 0.03	0.57 ± 0.04	0.78 ± 0.02c	0.57 ± 0.03a

^aP ≤ 0.05 compared to bisphenol A (BPA).

^bP ≤ 0.01 compared to BPA.

^cP ≤ 0.05 compared to control.

^dP ≤ 0.01 compared to control. Number of animals = 6 rats/group. Initial body weight, food consumption, and water intake = body weight, food consumption, and water intake at the first day (day 0) of the experiment. Final body weight, food consumption, and water intake = body weight, food consumption, and water intake at the final day of the experiment. Values are expressed as the mean ± SD. FA: Fertaric acid; BPA: Bisphenol A.

Table 2 Protective effect of fertaric acid on liver toxicity in rats exposed to bisphenol A

Parameter	Group
	Control	Paraffin oil	FA	BPA	FA + BPA
Serum AST (U/L)	121.8 ± 2.64	122.4 ± 3.26	124.0 ± 2.73	175.2 ± 0.72d	130.4 ± 4.17b
Serum ALT (U/L)	60.9 ± 2.37	61.9 ± 2.18	63.1 ± 2.59	86.7 ± 1.96d	65.1 ± 2.9b
Serum ALP (U/100 mL)	14.9 ± 1.27	14.7 ± 1.46	16.2 ± 1.56	5.65 ± 0.82d	13.7 ± 1.22b
Serum ACP (U/100 mL)	17.0 ± 2.79	16.2 ± 2.48	15.8 ± 2.75	6.86 ± 2.01d	12.15 ± 2.34b
Serum γGT (U/L)	8.42 ± 1.32	8.52 ± 1.25	9.90 ± 1.37	13.6 ± 2.30d	10.2 ± 1.57b
Serum LDH (U/L)	261.0 ± 43.6	257.9 ± 35.8	257.4 ± 36.8	755.9 ± 53.17d	277.8 ± 52.8b
Serum bilirubin (mg/dL)	0.53 ± 0.08	0.55 ± 0.09	0.58 ± 0.06	0.82 ± 0.05d	0.54 ± 0.04b

^aP ≤ 0.05 compared to bisphenol A (BPA).

^bP ≤ 0.01 compared to BPA.

^cP ≤ 0.05 compared to control.

^dP ≤ 0.01 compared to control. Number of animals = 6 rats/group. Values are expressed as the mean ± SD. FA: Fertaric acid; BPA: Bisphenol A; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; ALP: Alkaline phosphatase; ACP: Acid phosphatase; γ-GT: γ-Glutamyl transferase; LDH: Lactate dehydrogenase.

Table 3 Protective effect of fertaric acid on kidney toxicity and serum electrolytes in rats exposed to bisphenol A

Parameter	Group
	Control	Paraffin oil	FA	BPA	FA + BPA
Serum urea (mg/dL)	26.5 ± 2.83	27.2 ± 2.54	25.8 ± 2.59	34.0 ± 2.59c	28.2 ± 2.74a
Serum creatinine (mg/dL)	0.75 ± 0.08	0.76 ± 0.06	0.74 ± 0.07	0.98 ± 0.09c	0.76 ± 0.09a
Serum uric acid (mg/dL)	8.21 ± 0.43	8.16 ± 0.52	8.28 ± 0.64	17.65 ± 0.82d	9.16 ± 0.58b
Blood urea nitrogen (mg/dL)	16.25 ± 1.54	16.47 ± 1.39	16.18 ± 1.52	34.42 ± 1.80d	17.18 ± 1.71b
Serum sodium (mmol/L)	154.60 ± 3.29	155.28 ± 3.61	153.81 ± 3.50	110.24 ± 4.16d	144.9 ± 3.28a
Serum potassium (mmol/L)	5.62 ± 0.19	5.71 ± 0.15	5.59 ± 0.24	2.71 ± 0.28d	5.43 ± 0.18a
Serum chloride (mmol/L)	102.83 ± 2.19	103.20 ± 2.34	102.36 ± 2.75	81.47 ± 1.85d	101.26 ± 2.39a

^aP ≤ 0.05 compared to bisphenol A (BPA).

^bP ≤ 0.01 compared to BPA.

^cP ≤ 0.05 compared to control.

^dP ≤ 0.01 compared to control. Number of animals = 6 rats/group. Values are expressed as the mean ± SD. FA: Fertaric acid; BPA: Bisphenol A.

Table 4 Protective effect of fertaric acid on testicular toxicity in rats exposed to bisphenol A

Parameter		Group
	Control	Paraffin oil	FA	BPA	FA + BPA
Serum Ts (ng/mL)	5.98 ± 0.42	6.00 ± 0.51	5.96 ± 0.62	3.14 ± 0.49c	5.89 ± 0.68a
Serum LH (mIU/mL)	18.28 ± 1.85	18.31 ± 1.64	17.96 ± 1.93	33.30 ± 2.21d	18.15 ± 5.14b
Serum FSH (mIU/mL)	1.05 ± 0.13	1.03 ± 0.19	0.98 ± 0.15	2.32 ± 0.22d	1.02 ± 0.18b
Serum DHEA-SO4 (µg/dL)	197.50 ± 23.29	198.25 ± 26.12	195.74 ± 21.84	154.25 ± 13.56c	193.72 ± 19.71a
Serum SHBG (nmol/L)	6.65 ± 0.49	6.67 ± 0.62	6.63 ± 0.51	9.06 ± 1.84c	6.71 ± 4.95a
Testis G6PD (U/g tissue)	11.92 ± 0.66	11.94 ± 0.86	11.89 ± 0.73	5.64 ± 0.43d	10.86 ± 1.52b
Testis 3βHSD (U/g tissue)	4.46 ± 0.86	4.48 ± 0.75	4.43 ± 0.62	2.15 ± 0.36d	4.35 ± 0.88b
Testis Chol (mg/g tissue)	130.67 ± 8.16	132.17 ± 6.90	128.86 ± 7.48	192.53 ± 8.44d	129.21 ± 7.51b
Testis protein (mg/g tissue)	290.6 ± 14.23	288.9 ± 16.51	292.19 ± 13.64	187.60 ± 15.18d	289.45 ± 11.59b

^aP ≤ 0.05 compared to bisphenol A (BPA).

^bP ≤ 0.01 compared to BPA.

^cP ≤ 0.05 compared to control.

^dP ≤ 0.01 compared to control. Number of animals = 6 rats/group. Values are expressed as the mean ± SD. FA: Fertaric acid; BPA: Bisphenol A; Ts: Testosterone; LH: Luteinizing hormone; FSH: Follicle stimulating hormone; DHEA-SO₄: Dehydroepiandrosterone sulfate; SHBG: Sex hormone binding globulin; G6PD: Glucose-6-phosphate dehydrogenase; 3βHSD: 3β-hydroxysteroid dehydrogenase; Chol: Cholesterol.

Table 5 Liver DNA content in different groups

Control				BPA				FA + BPA
Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index
All	111	100.0%	1.000	All	106	100.0%	2.374	All	107	100.0%	1.923
5cER	0	0.0%	-	5cER	39	36.79%	2.948	5cER	11	10.28%	2.815
< 1.5c	24	21.62%	0.679	< 1.5c	0	0.0%	-	< 1.5c	0	0.0%	-
1.5c-2.5c	73	65.77%	1.031	1.5c-2.5c	24	22.64%	1.045	1.5c-2.5c	36	33.65%	1.181
2.5c-3.5c	13	11.71%	1.359	2.5c-3.5c	10	9.43%	1.558	2.5c-3.5c	17	15.89%	1.595
3.5c-4.5c	1	0.90%	1.779	3.5c-4.5c	33	31.13%	2.033	3.5c-4.5c	43	40.19%	2.273

Number of animals = 6 rats/group. The results are presented as a frequency histogram on the monitor generated by plotting the DNA content against the number of nuclei calculated. 2c: Diploid cells containing two copies of DNA; 3c: Proliferation index (S-phase cells containing three strands of DNA); 4c: Tetraploid cells containing four copies of DNA; > 4c: Cells with more than 4c DNA content; < 1.5c: Cells containing less than 1.5c DNA content. FA: Fertaric acid; BPA: Bisphenol A.

Table 6 Kidney DNA content in different groups

Control				BPA				FA + BPA
Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index
All	107	100.0%	1.000	All	106	100.0%	1.628	All	109	100.0%	1.136
5cER	0	0.0%	-	5cER	22	20.76%	2.716	5cER	8	7.34%	-
< 1.5c	13	12.15%	0.650	< 1.5c	0	16.04%	0.623	< 1.5c	0	8.26%	0.663
1.5c-2.5c	78	72.90%	0.984	1.5c-2.5c	21	19.81%	0.984	1.5- 2.5c	63	57.80%	1.008
2.5c-3.5c	16	14.95%	1.364	2.5c-3.5c	33	31.13%	1.364	2.5c-3.5c	32	29.36%	1.411
3.5c-4.5c	0	0.0%	-	3.5c-4.5c	30	28.30%	1.988	3.5c-4.5c	6	5.51%	1.842

Number of animals = 6 rats/group. The results are presented as a frequency histogram on the monitor generated by plotting the DNA content against the number of nuclei calculated. 2c: Diploid (cells containing two copies of DNA; 3c: Proliferation index (S-phase cells containing three copies of DNA); 4c: Tetraploid cells containing four copies of DNA; > 4 c: Cells with more than 4c DNA content; < 1.5 c: Cells containing less than 1.5 c DNA content. FA: Fertaric acid; BPA: Bisphenol A.

Table 7 Testis DNA content in different groups

Control				BPA				FA + BPA
Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index	Range	Total cells	% cells	DNA index
All	113	100.0	1.000	All	109	100.0	2.614	All	111	100.0	1.951
5cER	0	0.0	-	5cER	41	37.62	2.953	5cER	13	11.71	2.726
< 1.5c	23	20.35	0.662	< 1.5c	0	0.0	-	< 1.5c	0	0.0	-
1.5c- 2.5c	75	66.37	1.071	1.5c- 2.5c	26	23.85	1.805	1.5c- 2.5c	41	36.94	1.250
2.5c- 3.5c	14	12.39	1.412	2.5c- 3.5c	12	11.01	1.741	2.5c- 3.5c	25	22.52	1.803
3.5c- 4.5c	1	0.89	1.503	3.5c- 4.5c	30	27.52	2.019	3.5c- 4.5c	32	28.83	1.948

Number of animals = 6 rats/group. The results are presented as a frequency histogram on the monitor generated by plotting the DNA content against the number of nuclei calculated. 2c: Diploid cells containing two copies of DNA; 3c: Proliferation index (S-phase cells contained three copies of DNA); 4c: Tetraploid cells containing four copies of DNA; > 4 c: Cells with more than 4c DNA content; < 1.5 c: Cells containing less than 1.5 c DNA content. FA: Fertaric acid; BPA: Bisphenol A.