Codeine-induced sperm DNA damage is mediated predominantly by oxidative stress rather than apoptosis

Figures & data

Table 1. Effect of chronic codeine use on sperm parameters.

Variables	Control	Low-dose codeine	High-dose codeine
Sperm concentration (×106/ml)	67.14 ± 1.57^a	48.43 ± 4.42^b	43.00 ± 3.69^c
Sperm motility (%)	91.71 ± 2.36^a	67.57 ± 4.64^b	55.71 ± 5.99^c
Sperm viability (%)	85.43 ± 5.47^a	66.87 ± 6.09^b	52.71 ± 2.98^c

Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Table 2. Effect of chronic codeine use on sperm morphology.

Variables	Control	Low-dose codeine	High-dose codeine
Tailless head	3.86 ± 0.69^a	7.57 ± 1.39^b	10.43 ± 1.13^c
Headless tail	3.57 ± 0.97^a	7.43 ± 1.39^b	10.85 ± 0.89^c
Rudimentary tail	1.71 ± 0.95^a	7.28 ± 1.11^b	10.71 ± 1.38^c
Bent tail	5.86 ± 1.67^a	14.42 ± 0.97^b	15.00 ± 1.00^b
Curved tail	5.14 ± 1.21^a	12.28 ± 1.25^b	14.71 ± 0.95^c
Looped tail	2.00 ± 0.81^a	8.00 ± 0.81^b	10.71 ± 0.75^c
Bent midpiece	6.28 ± 1.49^a	13.00 ± 1.73^b	14.14 ± 1.21^b
Curved midpiece	7.71 ± 0.95^a	13.71 ± 1.70^b	15.28 ± 0.75^c
Percentage of abnormal sperm (%)	8.95 ± 1.02^a	23.46 ± 1.92^b	31.25 ± 2.05^c
Percentage of normal sperm (%)	91.02 ± 1.01^a	76.53 ± 1.92^b	68.75 ± 2.05^c

Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Table 3. Effect of chronic codeine use on sperm oxidative markers and anti-oxidants.

Variables	Control	Low-dose codeine	High-dose codeine
MDA (nmol/ml)	5.04 ± 0.14^a	7.08 ± −0.13^b	8.15 ± 0.17^c
MPO (U/mg)	54.52 ± 1.34^a	77.12 ± 2.39^b	86.25 ± 1.79^c
SOD (U/mg)	825.43 ± 4.31^a	786.43 ± 2.43^b	740.29 ± 3.40^c
GPx (U/mg)	420.00 ± 3.10^a	369.86 ± 6.86^b	320.10 ± 4.34^c

Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Figure 1. Effect of chronic codeine use on serum and testicular testosterone. Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Figure 2. Effect of chronic codeine use on sperm membrane integrity using HOST. Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Figure 3. Effect of chronic codeine use on sperm DFI, 80HdG, and caspase3. Same parameter carrying different alphabets (a,b,c) are statistically different at P < 0.05.

Table 4. Correlations observed between 8OHdG, caspase and sperm quality.

Variables	8OHdG	Caspase 3 activity	P value
Sperm concentration	−0.904	−0.836	0.000
Sperm motility	−0.934	−0.853	0.000
Sperm viability	−0.935	−0.857	0.000
Percentage of normal sperm	−0.961	−0.872	0.000
HOST at 30 min	−0.938	−0.879	0.000
HOST at 60	−0.942	−0.894	0.000
Sperm DNA Fragmentation	0.974	0.903	0.000

Table 5. Multivariate regression analysis to assess the association between oxidative DNA damage and apoptosis, and sperm DNA fragmentation.

	B	SE	Βeta	T	p
Sperm 8OHdG	3.405	0.464	0.835	7.343	0.000
Sperm caspase 3 activity	1.023	0.747	0.156	1.370	0.187

B: unstandardized regression coefficient; SE: standard error for the unstandardized regression coefficient; Beta (β): standardized regression coefficient; t: t test statistic; p: probability value.

Figure 4. Proposed mechanism of the effect of codeine on sperm cell.