Lipid peroxidation and decline in antioxidant status as one of the toxicity measures of diazinon in the testis

Figures & data

Table 1. Diazinon toxicity impacts on body and organ weight

Groups	1 week	2 weeks	8 weeks*
	Body weight (g)
Control	151.83 ± 10.92	159.17 ± 7.66	204.67 ± 19.09
10 mg/kg	156.00 ± 7.51	160.50 ± 11.88	203.00 ± 12.08
15 mg/kg	151.67 ± 9.24	150.83 ± 7.80	175.67 ± 12.75
30 mg/kg	135.00 ± 7.87	159.50 ± 11.74	175.67 ± 8.57
	Testis index (%)
Control	1.94 ± 46.75	1.80 ± 0.13	1.75 ± 0.15
10 mg/kg	1.96 ± 29.59	1.95 ± 0.13	1.80 ± 0.11
15 mg/kg	1.98 ± 31.22	1.90 ± 0.10	2.19 ± 0.10
30 mg/kg	2.08 ± 21.00	1.88 ± 0.16	2.32 ± 0.10**

Results are expressed as mean ± SEM (n = 6).

* Significant (P < 0.05) increase or decrease of DZN time-dependent toxicity effect when compared to the one week exposure group.

** Significant (P < 0.05) increase or decrease of DZN dose-dependent toxicity effect when compared to the control.

Table 2. Effects of diazinon on testicle antioxidants and pro-oxidant (lipid peroxidation) level

Groups	1 week	2 weeks	8 weeks*
Reduced glutathione (μmol/g tissue)
Control	6.52 ± 0.02	6.49 ± 0.16	6.31 ± 0.08
10 mg/kg	6.00 ± 0.20	6.15 ± 0.06	5.08 ± 0.06**
15 mg/kg	5.80 ± 0.09**	5.75 ± 0.13**	5.37 ± 0.07**
30 mg/kg	5.71 ± 0.02**	5.62 ± 0.18**	4.18 ± 0.09**
Catalase (μmol H2O2 consumed/min/mg protein)
Control	0.21 ± 0.03	0.23 ± 0.03	0.57 ± 0.27
10 mg/kg	0.16 ± 0.02	0.21 ± 0.03	0.56 ± 0.03
15 mg/kg	0.13 ± 0.02	0.19 ± 0.03	0.48 ± 0.36
30 mg/kg	0.12 ± 0.03	0.17 ± 0.02	0.38 ± 0.03**
Lipid peroxidation (nmol MDA formed/g tissue)
Control	2.37 ± 0.19	2.17 ± 0.16	2.03 ± 0.04
10 mg/kg	2.88 ± 0.11	3.49 ± 0.16**	4.17 ± 0.08**
15 mg/kg	2.57 ± 0.16	4.12 ± 0.19**	4.83 ± 0.13**
30 mg/kg	3.15 ± 0.21	5.02 ± 0.19**	6.14 ± 0.04**

Results are expressed as mean ± SEM (n = 6).

* Significant (P < 0.05) increase or decrease of DZN time-dependent toxicity effect when compared to the one week exposure group.

** Significant (P < 0.05) increase or decrease of DZN dose-dependent toxicity effect when compared to the control.

Table 3. The influence of diazinon on serum testosterone synthesis and lactate dehydrogenase activity

Groups	Testosterone (ng/ml)
	1 week	2 weeks	8 weeks
Control	1.40 ± 0.02	1.65 ± 0.08	1.75 ± 0.04
10 mg/kg	1.33 ± 0.02	1.60 ± 0.09	1.39 ± 0.03*
15 mg/kg	1.59 ± 0.04*	1.54 ± 0.10	1.32 ± 0.04*
30 mg/kg	1.53 ± 0.03*	1.15 ± 0.10*	1.12 ± 0.03*
	Lactate dehydrogenase (mIU/ml)
	1 week	2 weeks**	8 weeks**
Control	146.97 ± 11.05	147.82 ± 10.21	151.54 ± 9.56
10 mg/kg	143.92 ± 11.94	209.63 ± 8.56*	214.74 ± 10.08*
15 mg/kg	144.71 ± 12.26	219.76 ± 11.96*	216.06 ± 12.68*
30 mg/kg	144.36 ± 8.32	232.85 ± 9.78*	231.09 ± 12.73*

Results are expressed as mean ± SEM (n = 6).

*Significant (P < 0.05) increase or decrease of DZN time-dependent toxicity effect when compared to the control.

**Significant (P < 0.05) increase or decrease of DZN dose-dependent toxicity effect when compared to the one week exposure group.

Figure 1. Photomicrographs of testis sections of DZN-treated animals (1 week), (A) normal seminiferous tubules (L indicates lumen) in untreated animals, (B) shrinkage (*) of seminiferous tubules in 10 mg/kg b.w. DZN-treated group, (C) halo appearance or vacuoles (arrows) in the seminiferous tubules of 15 mg/kg b.w. DZN-treated group, and (D) more vacuoles (arrows) in the seminiferous tubules of 30 mg/kg b.w. DZN-treated group. Vacuoles were also observed in 2 weeks of DZN exposure – 10, 15, and 30 mg/kg groups. Slides were stained with haematoxylin-eosin dye. Scale, 200 µm.

Figure 2. Photomicrographs of testis sections of DZN-treated animals, (A) in 2 weeks of DZN exposure (15 and 30 mg/kg b.w.), treated animals showed disorganization and disintegration of cell, which caused secondary spermatocyte (arrows) moved into lumen (100×), (B) shrinkage (*) of seminiferous tubules and loss of cells (lc), (C) more immature cells appeared in the lumen of 30 mg/kg b.w. DZN-treated animals (arrows), and (D) abnormal formation of spermatozoa in the lumen. Slides were stained with haematoxylin-eosin dye. Scale of (A) and (B) – 200 µm, (C) and (D) – 100 µm.

Table 4. Diazinon induced gonadotoxicity effects

Groups	1 week	2 weeks	8 weeks*
Concentration (millions/ml)
Control	72.50 ± 2.14	67.50 ± 4.74	69.83 ± 4.23
10 mg/kg	68.00 ± 5.20	69.33 ± 4.56	95.67 ± 4.57
15 mg/kg	72.50 ± 5.86	57.67 ± 5.12	72.83 ± 5.59
30 mg/kg	64.50 ± 4.75	74.83 ± 4.78	87.00 ± 4.78
Abnormality (%)
Control	18.67 ± 0.94	14.75 ± 1.12	19.67 ± 0.56
10 mg/kg	22.08 ± 1.32	12.08 ± 1.11	24.17 ± 0.84**
15 mg/kg	16.75 ± 1.22	13.83 ± 1.17	28.33 ± 0.82**
30 mg/kg	21.67 ± 1.46	13.50 ± 1.37	29.83 ± 0.86**
Seminiferous tubular diameter (μm)
Control	520.51 ± 9.58	512.78 ± 12.33	548.95 ± 21.74
10 mg/kg	517.64 ± 12.31	539.53 ± 16.52	538.64 ± 13.94
15 mg/kg	513.92 ± 11.68	524.67 ± 15.23	533.90 ± 22.37
30 mg/kg	486.12 ± 8.23	502.46 ± 22.67	578.72 ± 20.89

Results are expressed as mean ± SEM (n = 6).

*Significant (P < 0.05) increase or decrease of DZN time-dependent toxicity effect when compared to the one week exposure group.

** Significant (P < 0.05) increase or decrease of DZN dose-dependent toxicity effect when compared to the control.

Figure 3. Photomicrographs show abnormal sperms, (A) clumping, (B) head deformity (arrow), (C) hookless (a) and coiled tail (b), (D) bent-tail (arrow, (E) broken-head (a) neck's defect (n), (F) broken head (a), broken tail (b) and bent tail (arrow), (G) double head (arrow). Slides were stained with haematoxylin-eosin dye. Scale, 100 µm.