Ethanolic extract of propolis and vitamin E attenuates metal-induced testicular necrosis: time-related study on male reproductive system in albino mice

Figures & data

Table I. Serum Follicle stimulating hormone (FSH ng/mL), Luteinizing hormone (LH ng/mL) and testosterone (ng/mL) of male albino mice in Control (I), Al (II), vitamin E (III), propolis (IV), Al+vitamin E (V), Al+propolis (VI), Al+vitamin E+propolis (VII) groups after 1^st, 2^nd and 3^rd samplings

Parameters	Sampling	I	II	III	IV	V	VI	VII
FSH (ng/mL)	1^st	90.16 ± 3.77	72.22 ± 2.78	102.11 ± 2.49+#	100.11 ± 3.70^+##	103.31 ± 3.03^+##	70.44 ± 3.68^##	64.16 ± 1.62^**##
	2^nd	78.05 ± 3.88	61.71 ± 1.23	72.18 ± 3.16	73.67 ± 2.46	72.48 ± 2.10	69.38 ± 2.61	75.66 ± 6.01
	3^rd	78.40 ± 2.01	48.38 ± 2.88^**@+	98.18 ± 0.74^#	93.62 ± 3.71^##	90.18 ± 0.38^**##	63.78 ± 2.30^##	77.56 ± 2.79^##
LH (ng/mL)	1^st	26.38 ± 2.88	3.46 ± 0.18^**@@	16.38 ± 0.88	14.93 ± 0.35	4.52 ± 0.19	3.40 ± 0.20	3.84 ± 0.31
	2^nd	26.63 ± 0.47	0.84 ± 0.17	11.36 ± 0.24	13.80 ± 0.16	2.58 ± 0.63	2.63 ± 0.53	9.27 ± 0.62
	3^rd	23.18 ± 1.56	0.07 ± 0.01^*++@@	14.49 ± 0.30^*++	14.09 ± 0.38^*++	2.88 ± 0.33^*++@@	3.20 ± 0.09^*++@@	5.89 ± 0.33^*+@@
Testosterone (ng/mL)	1^st	1.14 ± 0.1	2.16 ± 0.16*	1.88 ± 0.08	1.90 ± 0.19	2.36 ± 0.13	2.13 ± 0.14	1.81 ± 0.04
	2^nd	1.82 ± 0.09	0.84 ± 0.03	1.74 ± 0.12	2.43 ± 0.04	2.05 ± 0.05	2.22 ± 0.09	2.67 ± 2.67 ± 0.03
	3^rd	1.60 ± 0.02	0.581 ± 0.03^**++@@#	2.08 ± 0.13^+##*@	2.05 ± 0.13^+##*@	2.27 ± 0.12^+##@	2.171 ± 0.10^+#	1.85 ± 0.03^+@@##

* = versus Control 1 W1; @ = versus Control W3; + = versus AlCl₃ W1; # = versus AlCl₃ W3.

*, @, +, # = p < 0.05; **, @@, ++, ## = p < 0.01.

Table II. Time-related effects of different treatments on absolute and relative weight of male reproductive organs after 1^st, 2^nd and 3^rd samplings

Parameters	Sampling	I	II	III	IV	V	VI	VII
Testis absolute weight (g)	1^st	0.2744 ± 0.02	0.2732 ± 0.01	0.2777 ± 0.01	0.2768 ± 0.01	0.2750 ± 0.01	0.2755 ± 0.01	0.2756 ± 0.01
	2^nd	0.2752 ± 0.01	0.2729 ± 0.02	0.2765 ± 0.01	0.2770 ± 0.01	0.2762 ± 0.02	0.2750 ± 0.01	0.2760 ± 0.01
	3^rd	0.2885 ± 0.01	0.2591 ± 0.02*	0.2748 ± 0.01	0.2754 ± 0.02	0.2778 ± 0.01^a	0.2743 ± 0.01^a	0.2895 ± 0.01^aa
Testis relative weight (g)	1^st	1.1409 ± 0.02	0.9076 ± 0.01	1.6192 ± 0.01	1.4794 ± 0.02	1.1574 ± 0.01	1.0715 ± 0.02	1.24200 ± 0.01
	2^nd	1.0576 ± 0.01	0.9000 ± 0.01	1.5134 ± 0.01	1.3454 ± 0.01	1.1000 ± 0.02	1.0654 ± 0.01	1.20000 ± 0.02
	3^rd	0.9584 ± 0.01	0.7750 ± 0.01*	1.1668 ± 0.03^a	1.0791 ± 0.01^a	1.0025 ± 0.02^a	0.8877 ± 0.01	1.1344 ± 0.01^a
Epididymis and vas deferens – absolute weight (g)	1^st	0.0867 ± 0.01	0.0789 ± 0.03	0.0815 ± 0.01	0.0875 ± 0.04	0.0787 ± 0.01	0.0804 ± 0.03	0.0810 ± 0.01
	2^nd	0.0865 ± 0.02	0.0749 ± 0.01	0.0817 ± 0.01	0.0870 ± 0.01	0.0781 ± 0.02	0.0810 ± 0.01	0.0823 ± 0.02
	3^rd	0.0864 ± 0.01	0.0733 ± 0.03	0.0822 ± 0.02	0.0863 ± 0.03	0.0790 ± 0.01	0.0827 ± 0.03	0.0850 ± 0.03
Epididymis and vas deferens – relative weight (g)	1^st	0.3604 ± 0.01	0.26212 ± 0.02	0.47521 ± 0.01	0.46766 ± 0.02	0.33122 ± 0.02	0.31271 ± 0.02	0.3650 ± 0.01
	2^nd	0.3500 ± 0.01	0.25431 ± 0.01	0.47200 ± 0.02	0.46500 ± 0.01	0.3132 ± 0.01	0.31170 ± 0.01	0.3600 ± 0.01
	3^rd	0.2877 ± 0.01	0.20242 ± 0.03	0.34904 ± 0.03^a	0.3381 ± 0.01^a	0.2850 ± 0.01	0.26763 ± 0.01^a	0.3330 ± 0.02*^aa
Seminal vesicle – absolute weight (g)	1^st	0.3857 ± 0.08	0.2770 ± 0.01	0.3842 ± 0.09	0.3456 ± 0.05	0.3265 ± 0.04	0.3331 ± 0.01	0.3764 ± 0.07
	2^nd	0.3868 ± 0.01	0.2781 ± 0.01	0.3820 ± 0.05	0.3567 ± 0.01	0.3231 ± 0.01	0.3212 ± 0.01	0.3700 ± 0.02
	3^rd	0.3736 ± 0.07	0.2536 ± 0.02**	0.3794 ± 0.01^a	0.3756 ± 0.04^a	0.3655 ± 0.03^a	0.3129 ± 0.01^a	0.3850 ± 0.01^aa
Seminal vesicle – relative weight (g)	1^st	1.6037 ± 0.07	0.92026 ± 0.01	2.240 ± 0.09	1.8471 ± 0.02	1.37415 ± 0.03	1.29560 ± 0.01	1.69625 ± 0.03
	2^nd	1.5600 ± 0.03	0.978 ± 0.01	2.143 ± 0.02	1.7356 ± 0.01	1.36400 ± 0.01	1.1789 ± 0.01	1.64567 ± 0.01
	3^rd	1.2411 ± 0.06	1.05011 ± 0.02*	2.0277 ± 0.08*	1.4717 ± 0.05	1.3190 ± 0.04	1.01262 ± 0.01	1.50862 ± 0.01*^a

* = versus Control; a = versus AlCl₃ # = p < 0.05; **, aa = p < 0.01; C (I), Al (II), vitamin E (III), propolis (IV), Al+vitamin E (V), Al+propolis (VI), Al+vitamin E+propolis (VII).

Table III. Comparison of sperm count and motility between the control and treatment groups after 1^st, 2^nd and 3^rd samplings

Parameters	Weeks	I	II	III	IV	V	VI	VII
Sperm count (× 10^16)	1^st	6.760 ± 0.835	6.598 ± 0.957	6.676 ± 0.754	6.659 ± 0.680	6.533 ± 0.571	6.635 ± 0.546	6.732 ± 0.765
	2^nd	6.730 ± 0.876	6.0678 ± 0.878	6.579 ± 0.654	6.630 ± 0.620	6.477 ± 0.587	6.367 ± 0.568	6.656 ± 0.76
	3^rd	6.859 ± 0.893	3.788 ± 0.678**	6.316 ± 0.557	6.534 ± 0.470	4.851 ± 0.790	4.511 ± 0.403	5.980 ± 0.654 ^s
Sperm motility	1^st	76.510 ± 8.212	60.954 ± 7.851*	74.157 ± 7.985	76.427 ± 7.256	74.558 ± 6.652	75.408 ± 7.901	76.332 ± 6.780
	2^nd	75.765 ± 0.786	58.98 ± 7.671	75.100 ± 7.876	75.456 ± 7.345	72.8764 ± 6.435	74.402 ± 7.657	75.200 ± 5.654
	3^rd	78.438 ± 0.934	50.238 ± 6.980**	77.157 ± 7.985	71.110 ± 5.650	69.163 ± 7.087	70.763 ± 9.643^a	77.442 ± 9.901^aa

* = versus Control; a = versus AlCl₃ # = p < 0.05; **, aa = p < 0.01; C (I), Al (II), vitamin E (III), Propolis (IV), Al+vitamin E (V), Al+propolis (VI), Al+vitamin E+propolis (VII).

Figure 1. (a) Control group during 1^st and 3^rd week of treatment showing normal structure at (400x)Spermatid population (Sp P), lumen (L), seminiferous tubules (S T), basement membrane (BM),spermatocytes (Sp), interstitial cells (I C), interstitial spaces (I S), tunica albuginea (T A). (b) Testes ofmice treated with AlCl₃ for 1 week showing damaged testicular tissues at (100x) Interstitial spaces (I S),lumen (L), blood vessels (B V), seminiferous tubules (S T), shrunken interstitial cells (S I C), damagedbasement membrane (D B M). (c) Cross section of testis of male albino mice treated with AlCl₃ for twoweeks showing damaged testicular tissues at (100x), (H & E). Tunica albuginea (T A), irregular shapedseminiferous tubules (Ir S T), interstitial spaces (I S), basement membrane (B M), interstitial cells (I C),Information Classification: Generaland spermatocytes (Sp). (d) Testes of mice treated with AlCl₃ for three weeks (400x) Vacuolation (V),damaged spermatocytes (Sp), lumen (L), interstitial cells (I C), irregular shaped seminiferous tubules (ir ST), basement membranes (B M). (e) Testes of mice treated with Vitamin E for 1 week (400x)Seminiferous tubules (S T), lumen (L), interstitial cells (I C), spermatocytes (Sp). (f) Cross section oftestes of vitamin E treated male albino mice for two weeks showing normal structure at (400x), (H & E).Spermatid population (Sp P), basement membrane (B M), interstitial cells (I C), lumen (L), spermatocytes(Sp). (g) Testes of mice treated with vitamin E for three weeks (400x), (H & E) Basement membranes(BM), interstitial cells (I C), lumen (L), spermatid population (Sp P). (h) Testes of mice treated for oneweek with propolis (400x)

Figure 1. (Continued). (i) Cross section of testes of male albino mice treated with propolis for twoweeks showing normal structure at (400x), (H & E). Spermatocytes (Sp), interstitial cells (I C), basementmembranes (B M), seminiferous tubules (S T). (j) Testes of mice treated with propolis for three weeks(400x), (H & E). Lumen (L), basement membrane (B M), interstitial cells (I C), interstitial spaces (I S),seminiferous tubules. (k) Testes of mice treated with AlCl₃ + vitamin E after 1 week (100x). Interstitialcells (I C), irregular shaped seminiferous tubules (Ir S T), damaged seminiferous tubules (D S T), bloodvessels (B V), interstitial spaces (I S), seminiferous tubules regular in shape (R S T), leydig cells (L C). (l)Cross section of testes of male albino mice showing effects after treatment with AlCl₃ and vitamin E fortwo weeks at (100x), (H & E). The structure of seminiferous tubules (S T), lumen of the S T (L),interstitial spaces (I S). (m) Testes of mice showing effects after treatment with AlCl₃ + vitamin E forthree weeks (100x). Seminiferous tubules (S T), basement membranes (B M), lumen having cells in it(L(c)), interstitial spaces (I S), interstitial cells (I C). (n) Testes of mice treated with AlCl₃ and propolisfor 1 week (100x). Intercellular spaces (I S), decreased lumen (De L), basement membranes (B M),=irregular seminiferous tubules (Ir S T), contracted interstitial cells (I C). (o) Cross section of testes ofmale albino mice showing effects after treatment with AlCl₃ and propolis for two weeks at (100x), (H & E). Tunica albuginea (T A), damaged seminiferous tubules (D S T), interstitial cells (I C), blood vessels(B V). (p) Testes of mice treated with AlCl₃ and propolis for three weeks (100x), (H & E). Seminiferoustubules (S T), interstitial cells (I C), blood vessels (B V), leydig Cells (L C)

Figure 1. (Continued). (q) Testes of mice treated with AlCl₃, vitamin E and propolis for one week at (400x). Interstitial spaces (I S), spermatocytes (Sp), seminiferous tubules (S T), tunica albuginea (T A). (r) Cross section of testes of male albino mice showing effects after treatment with AlCl₃, vitamin E and propolis for two weeks at (400x), (H & E). Spermatocytes (Sp), interstitial spaces (I S), leydig cells (L C), basement membranes (B M), seminiferous tubules (S T). (s) Testes of mice with AlCl₃, vitamin E and treated with propolis for three weeks (400x). Spermatid population (Sp P), spermatocytes (Sp), interstitial cells (I C), interstitial spaces (I S), basement membranes (B M), lumen (L), seminiferous tubules (S T)