Therapeutic and fertility restoration effects of Ionidium suffruticosum on sub-fertile male albino Wistar rats: effects on testis and caudal spermatozoa

Figures & data

Table 1. Effect of I. suffruticosum on body and testis weight and epididymal spermcounts in Carbendazim-induced subfertile male rats after 28 days of treatment.

	Body weight (g)
Experimental group	Initial ± SEM	Final ± SEM	Testis weight(mg/kg body wt) ± SEM	Caudal epididymalsperm counts × 10^6 sperm/ml ± SEM
Group 1: Control (Saline only)	164 ± 0.6	172 ± 0.7	1248 ± 1.4	40.50 ± 0.5
Control (extract only)	164 ± 0.7	174 ± 0.4	1258 ± 1.2	49.0 ± 0.7
Group 2: Carbendazim-induced subfertile male rats	158 ± 0.7	125 ± 0.7	851 ± 0.3	15.40 ± 0.3
Group 3: Carbendazim-induced subfertile male rats treated with I. suffruticosum	156 ± 0.4*	132 ± 0.6*	948 ± 0.4*	49.60 ± 0.5*
Group 4: Carbendazim-induced subfertile male rats treated with Clomiphene citrate	150 ± 0.7	120 ± 0.5	906 ± 0.3	45.50 ± 0.4

Each value is of Mean ± SEM of five animals.

* Significantly different (p < 0.05) from the control group by Dunnett’s test.

Scheffé multiple comparison tests used to identify the pairs of treatments are significantly different from each other. Control saline vs control extract, Group 2 vs Group 3 in initial body weight, control saline vs control extract in final body weight and in caudal epididymal sperm counts Group extract vs Group 3 are statistically insignificant (p > 0.01) and in testis weight all groups are statistically significant (p < 0.01) .

Table 2. Effect of I. suffruticosum on Cauda epididymal sperm motility and sperm pattern in Carbendazim-induced subfertile male rats after 28 days of treatment.

Experimental groups	Cauda epididymalsperm motilityand pattern (Min)
Group 1: Control (saline only)	55.17 ± 0.4
Control (extract only)	60.12 ± 0.7
Group 2: Carbendazim-induced subfertile male rats	10.16 ± 1.8
Group 3: Carbendazim-induced subfertile male rats treated with I. suffruticosum	56.20 ± 0.8*
Group 4: Carbendazim-induced subfertile male rats treated with Clomiphene citrate	57.11 ± 0.2

Each value is Mean ± SEM of five animals.

* Significantly different (p < 0.05) from the control group by Dunnett’s test.

Scheffé multiple comparison tests used to identify the pairs of treatments are significantly different from each other. Control saline vs Group 3, control saline vs Group 4, Group 3 vs Group 4 are statistically insignificant p > 0.01.

Table 3. Effect of Ionidium suffruticosum on Cauda epididymal sperm vitality and spermmorphology in Carbendazim induced subfertile male rats after 28 days of treatment.

Experimental groups	Sperm vitality		Sperm morphology
	Live sperms (%)	Dead sperms (%)	Normal sperms (%)	Abnormal sperms (%)
Group-1: Control (saline only)	83 ± 0.6	17 ± 0.5	93 ± 0.5	7 ± 0.8
Control (extract only)	91 ± 0.5	9 ± 0.3	97 ± 0.5	3 ± 0.3
Group 2: Carbendazim inducedsubfertile male rats	57 ± 0.4	43 ± 0.4	58 ± 0.5	42 ± 0.5
Group 3: Carbendazim induced subfertile male rats treated with Ionidium suffruticosum	85 ± 0.4*	15 ± 0.4*	90 ± 0.7*	10 ± 0.3*
Group 4: Carbendazim induced subfertile male rats treated with clomiphene citrate	83 ± 0.4	17 ± 0.3	87 ± 0.4	13 ± 0.7

Each value is of Mean ± SEM of 5 animals.

* Significantly different (p < 0.05) from the control group by Dunnett’s test.

Scheffé multiple comparison tests used to identify which of the pairs of treatments are significantly different from each other. Control saline Vs Group 4 in live sperms, Control saline Vs Group 3, Control saline Vs Group 4, Group 3 Vs Group 4, in dead sperms are statistically insignificant p > 0.01 and in normal and abnormal sperms counts all groups are statistically significant p < 0.01.

Table 4. Percentage of predominant abnormalities of sperm observed in different experimentalgroups.

Experimental Groups	Predominant abnormalities (%)
	TFFAF	DTH	FAS	BMP	DTT	COF	TCOF
Group-1
Saline only	2 ± 0.3	3 ± 0.3	1 ± 0.3	0 ± 0	1 ± 0.4	0 ± 0	0 ± 0
Extract only	1 ± 0.3	0 ± 0	0.5 ± 0.1	0 ± 0	1 ± 0.3	0 ± 0	0.5 ± 0.1
Group-2	10 ± 0.7	13 ±o.7	9 ± 0.3	6 ± 0.5	2 ± 0.2	1 ± 0.3	1 ± 0.3
Group-3	2 ± 0.3*	4 ± 0.3*	0 ± 0*	1 ± 0.3*	1 ± 0.4*	0 ± 0*	1 ± 0.3*
Group-4	3 ± 0.4	5 ± 0.3	1 ± 0.3	2 ± 0.3	2 ± 0.3	0 ± 0	0 ± 0

Each value is of Mean ± SEM of 5 animals.*Significantly different (p < 0.05) from the control group by Dunnett’s test.

TFFAF: Sperm with head flexed, tip of the head facing away from the flagellum; DTH: Detached Head; FAS: Fusion/attachment of sperm at one or more points between two or more sperm. BMP: Broken middle piece; DTT: Detached Tail; COF: Coiling of the flagellum; TCOF: Twisting and coiling of the flagellum.

[Scheffé multiple comparison tests used to identify which of the pairs of treatments are significantly different from each other. Control saline Vs extract, Control saline Vs Group 3, Control saline Vs Group 4, Control extract Vs Group 3 in TFFAF, Control saline Vs Group 3, Group 3 Vs Group 4 in DTH, Control saline Vs extract, Control saline Vs Group 3, Control extract Vs Group 3, Group 3 Vs Group 4 BMB, Control saline Vs Group 2, Control extract Vs Group 2, Group 2 Vs Group 3, Group 2 Vs Group 4 in COF and all groups from DTT are statistically insignificant p > 0.01 and Control saline Vs Group 2, Control saline Vs Group 3, Control saline Vs Group 2, Control extract Vs Group 2, Group 2 Vs Group 3, Group 2 Vs Group 4 in FAS groups are statistically significant p < 0.01.

Figure 1. Effect of I. suffruticosum on epididymal sperm agglutination in Carbendazim-induced subfertile male rats after 28 days of treatment. (A) Sperms of Group of rats treated with I. suffruticosum (450×). (B) Sperms of Group of rats treated with Clomiphene citrate (450×). (C) Agglutination of sperms in carbendazim-treated group (450×). (D) Agglutination of sperms of group 3 rat treated with I. suffruticosum (450×). (E) Seminiferous tubule of a carbendazim-treated rat showing specific depletion of pachytenespermatocyte and total seminiferous tubular atrophy (450×). (F) Seminiferous tubules of a carbendazim-induced subfertile male rats treated with I. suffruticosum showing less damage and with a normal histology (100×). (G) Seminiferous epithelium of a carbendazim-treated rats showing sloughing off germinalepithelium (100×). (H) Single seminiferous tubules, of a group 3 rat treated with I. suffruticosum (450×) (I) Seminiferous tubules of a carbendazim-treated rat showing fibrosis (450×). (J) Single seminiferous tubules treated with I. suffruticosum showing normal histological restoration (450×) 1: Control; 2: Carbendazim-Treated; 3: I. suffruticosum treated; 4: Standard fertility drug Clomiphene citrate treated+

Figure 1. Continued

Table 5. Effect of Ionidium suffruticosum on malondialdehyde and antioxidant enzymelevels in Carbendazim induced subfertile male rats after 28 days of treatment.

Experimental groups	Enzyme levels
	MAD nmol/ml	CAT μmol/S/ml	SOD μ/ml
Group-1: Control (saline only)	27.9 ± 0.5	4.2 ± 0.6	136.2 ± 0.4
Control group (extract only)	28.2 ± 0.3	4.4 ± 0.3	138.3 ± 0.5
Group 2: Carbendazim induced sub fertilemale rats	42.3 ± 0.5	2.1 ± 0.1	88.7 ± 0.3
Group 3: Carbendazim induced subfertile male rats treated with Ionidium suffruticosum	33.6 ± 0.2*	3.8 ± 0.2*	131.3 ± 0.5*
Group 4: Carbendazim induced subfertile male rats treated with Clomiphene citrate	32.83 ± 0.3	3.6 ± 0.3	133.1 ± 0.5

MAD: Malondialdehyde; CAD: Catalase; SOD: Superoxide dismutase.

Each value is of Mean ± SEM of 5 animals.

*Significantly different (p < 0.05) from the control group by Dunnett’s test.

Scheffé multiple comparison tests used to identify which of the pairs of treatments are significantly different from each other. Control saline Vs Control extract, Group 3 Vs Group 4 in MAD, SOD are statistically insignificant p > 0.01 and in CAT Control saline Vs Group 2 and Control extracts Vs Group 2 are statistically significant p < 0.01.