Effect of smoking on semen quality, FSH, testosterone level, and CAG repeat length in androgen receptor gene of infertile men in an Indian city

Figures & data

Table 2. Characteristics of semen, reproductive hormones for participants according to level of current smoking.

Seminal characteristics	Non-smoker	Light smokers (1-20 cigarettes/day)	Heavy smokers (>20 cigarettes/day)
Seminal phenotype
Normozoospermic	76	49	12
Aspermic	2	14	18
Oligozoospermic	11	6	16
Severe oligozoospermic	8	4	4
Teratozoospermic	24	16	24
Asthenozoospermic	5	9	6
Total	126	98	80
Sperm motility
Nonmotile	5	9	6
Slow progressive motile	40	35	38
Rapid progressive motile	71	36	14
Total	116	80	58
Sperm morphology
Normal	76	36	15
Head defect	12	7	22
Tail defect	11	9	7
Multiple defect	17	28	14
Total	116	80	58
Sperm DNA integrity
Normal (> 50%)	88	46	33
Damaged DNA	28	34	25
Total	116	80	57

Table 3A. Logistic regression analysis of the confounding effect of age and BMI on seminal phenotype and smoking.

Predictor	Coef	OR (95%CI)	P value
Logit 1: (5/0)
Smoking	0.7091	2.03(1.04-3.98)	0.038
Age	−0.00471	1.00(0.89 − 1.12)	0.936
BMI	−0.14478	0.87(0.75 − 1.00)	0.050
Logit 2: (4/0)
Smoking	0.7844	2.19(1.42 − 3.38)	0.000
Age	0.00563	1.01(0.93 − 1.08)	0.881
BMI	0.00520	1.01(0.92 − 1.10)	0.907
Logit 3: (3/0)
Smoking	0.1020	1.11(0.53 − 2.33)	0.788
Age	0.06141	1.04(0.92 − 1.17)	0.528
BMI	0.07929	0.89(0.76 − 1.04)	0.142
Logit 4: (2/0)
Smoking	1.0448	2.84(1.64 − 4.92)	0.000
Age	−0.00482	1.00(0.91 − 1.09)	0.918
BMI	0.00530	1.01(0.90 − 1.12)	0.924
Logit 5: (1/0)
Smoking	1.4228	4.15(2.26 − 7.63)	0.000
Age	−0.01342	0.99(0.90 − 1.08)	0.781
BMI	−0.17059	0.84(0.74 − 0.95)	0.007

Phenotypes: 0(Reference Event)normozoospermic, 5 asthenozoospermic, 4 teratozoospermic, 3 severe oligozoospermic, 2 oligozoospermic, 1 azoozoospermic

Table 3B. Logistic regression analysis of sperm motility, morphology, and DNA integrity with smoking habit.

Logit/Parameter	Predictor	Coeff.	p-value	OR (95%CI)
Sperm motility	Constant Smoking habit	−1.1493 −0.7045	0.000 0.000	0.49(0.36 − 0.68)
Sperm morphology	Constant Smoking habit	−1.4897 0.8467	−0.000 0.000	2.33 (1.66-3.27)
DNA integrity	Constant Smoking habit	−1.4939 0.4638	0.000 0.008	1.59 (1.15-2.21)

Reference level: normo

Figure 1.  Assessment of human sperm DNA integrity by acridine orange staining (40X). DNA integrity was assessed using acridine orange staining following sperm fixation. The percentage of green (normal DNA integrity) and orange red (abnormal DNA integrity) spermatozoa per 200 spermatozoa in each sample was calculated. An abnormal integrity of sperm DNA was considered as more that 30% denaturation (orange-red).

Figure 2.  FSH, LH, and testosterone levels among non-smokers and smokers. Follicle stimulating hormone (FSH), leutanising hormone (LH), and testosterone concentrations were determined in the serum of smokers (severeoligo/azoospermic) and nonsmokers (normozoospermic). Values are mean and SD from two independent experiments. mIU, mili International Unit

Figure 3.  CAG repeat length in the AR genes of subjects. The CAG repeat segment of the AR gene was amplified by PCR with primers and conditions described in Materials and Methods. The representative products analyzed by agarose gel electrophoresis are shown. L1 and L2 represent repeat numbers 33 and 16, respectively. M, 500 base pair DNA size marker; AR: androgen receptor

Table 4. Distribution of length of CAG repeats in the androgen receptor gene.

ID	Age	BMI (kg/m^2)	FSH (mIU/ml)	LH (mIU/ml)	Testosterone (ng/ml)	CAG Repeat in AR gene
N287	34	22.2	5.8	5.2	5.6	14
N40	40	23.1	7.2	6.8	5.2	22
N7	39	19.8	6.8	7.7	6.2	22
N24	38	25.5	7.7	6.2	4.2	29
N160	36	24.5	5.5	5.8	6.2	22
N304	38	26.7	7.4	6.6	5.5	22
N251	41	22.1	7.2	5.8	6.7	16
N216	35	19.9	6.5	6.6	5.6	16
N139	36	22.2	8.2	7.8	4.8	14
N128	40	20.5	5.6	5.2	5.4	14
Mean ± SD	37.7 ± 2.35	22.6 ± 2.3	6.79 ± 0.9	6.37 ± 0.9	5.54 ± 0.7	19.1 ± 5.04
A70	39	20.1	10	9.8	1.6	29
A82	42	17.6	11.5	10.6	1.9	22
A8	40	20.2	12.3	10.2	1.5	29
A30	36	19.2	12	11.2	3.2	16
A146	39	19.7	10.6	9.2	1.0	33
A113	42	20.4	11.2	10.6	2.2	22
A238	39	23.1	11.5	10.5	1.5	32
A298	43	28.5	9.8	7.8	1.2	29
A171	34	18.5	10.4	8.8	1.8	29
A195	41	20.2	10.5	7.8	2.0	19
Mean ± SD	39.5 ± 2.7	20.75 ± 3.07	10.98 ± 0.84	9.65 ± 1.2	1.79 ± 0.6	26 ± 5.8
			P < 0.0001	P < 0.0001	P < 0.0001	P = 0.033

N: non-smoking group

A: smoking group

Table 1. Demographic status of the population included in the present study.

				P value
Parameters	Non smokers (n = 126)	Light smokers (n = 98)	Heavy smokers (n = 80)	NS/LS	NS/HS	LS/HS
Age (Mean ± SD)	35 ± 2.1	40 ± 5.2	41 ± 1	0.001	0.001	0.0921
BMI	25.92 ± 4.77	25.68 ± 5.92	21.31 ± 4.22	0.737	0.001	0.001
No. of cigarette/day	0	1 - 20	> 21
Sperm count x10^6 /ml	130 ± 50	76 ± 50	50 ± 20	0.001	0.001	0.001

NS: Non smoker

LS: Light smoker

HS: Heavy smoker