ABSTRACT- Background: Plant Cassia tora has been used in traditional and modern medicines for different pharmacological activities.
Objectives: The present investigation has been taken to observe and evaluate effects of Cassia tora on the reproduction functions of male rats in search a safe, orally effective and reversible fertility regulating agent.
Materials and Methods: Fifty percent ethanolic extract of Cassia tora was prepared and administered orally in male Wistar rats at the doses of 50, 100 and 200 mg/kg.b.wt./rat/day dose levels respectively for a period of 60 days and some of the treated rats were kept 30 days for recovery of fertility to assessed reversibility effects. Hematological indices, serum clinical investigations were also performed to assess toxic effects if any caused in rats by treatment. Proteins, cholesterol, glycogen, ascorbic acid, sialic acid and fructose level were analyzed in rats. Serum FSH, LH and Testosterone levels were measure. Rats were castrated to evaluate effects on reproductive functions of hormones and mode of action of the Cassia tora treatment. For histopathological observations tissues were fixed in Bouin’s fluid, dehydrated, sectioned and stained with Hematoxylin and Eosin.
Results: Treatment of Cassia tora significantly reduced the weights of testes and accessory sex organs. Sperm density and motility were declined high significantly. Levels of Testosterone and FSH hormone were significantly decreased in rats. The protein, sialic acid, fructose, ascorbic acid and glycogen contents of reproductive accessory sex organs were decreased significantly. Germinal epithelium of testes degenerated and number of spermatocytes, spermatids and spermatozoa in lumen of seminiferous tubules reduced.
Conclusions: The decreased testes and accessory sex organs weights, sperm motility, density and testosterone level in rats might be due to androgen suppression effects of Cassia tora treatment cause inhibition of spermatogenesis resulted reduction of fertility in treated male rats.
Key words: Cassia tora, Contraception, Fertility, Sperm motility, Sperm density, Male rat

INTRODUCTION
The population explosion creates immense pressure on our natural and non-renewable resources leading to social economic imbalances and political tension [1-2]. Fertility regulation has therefore, become a major global concern [3-5]. Although, there are several types of contraceptive methods are available for fertility control in male and females, but all of these were find one or more side effects. Survey of literature enumerate that the use of plants products as antifertility agent cause minimal or no side effects as compared to currently available conventional contraceptive methods especially oral. Literature reveals that the plants and their products were used for fertility regulation since hazards of uncontrolled population were visualized [6-9].
World Health Organization have already been taken significant steps to carry out research aimed at finding new and effective antifertility agents from traditional medicinal plants [10-11].
The results obtained are encouraging and thus hoped that in near future an easily administrable and reversible oral contraceptive of plant origin would be available to common people. The present investigation is planned to evaluate contraceptive efficacy of Cassia tora fruits products to search a cheap, orally effective and reversible contraceptive from indigenous medicinal plants with emphasis of mode of action and side effects in rats with the objective to develop a cheap, safe, easily administrable, orally effective and reversible male fertility regulating agents from traditional medicinal plants.

MATERIALS AND METHODS
The Plant- The plant Cassia tora is belongs to family- Fabaceae and also known as Cassia obstusifolia, Foetid Cassia, Sickle senna, Wild senna, Charota, Chakvat, Chakvat senna tora etc [12] have selected for the present study on the basis of literature survey of ethanomedicinal properties. The fruits of the plant were collected in the moths of September and October of 2010 and study was carried out at Department of Zoology, University of Rajasthan Jaipur, India.
Cassia tora possess various medicinal and pharmacological activities these includes antihepatotoxic [13], antiallergic [14], antimutagenic [15]., antifungal [16], radical scavenging and antimicrobial [16-25] .The chemical constituent of the plant Cassia tora are rubrofusarin triglucoside, non rubrofusarin gentobioside, demethyl-flavasperone, genitobioside, torachrysone gentibioside, torachrysone tetraglucoside, tora chrysone apioglucoside, torachrysone, toralactone aloemodin, rhein, emodin, naphthalene, anthraquinone, methicillin-resistant [23-26].

Preparation of Test material- Plant Cassia tora (RUBL20672) had been identified by deposit herbarium specimen stem at the Department of Botany, University of Rajasthan Jaipur, India. The fresh fruits of the plant Cassia tora were collected around the Jaipur district in the months of September and October of 2010.washed, shade dried and crushed to make fine powder for further use. The 50% alcoholic extract of fresh fruits of this plant was prepared according to WHO protocol CG-04 [27].

Animal model- Colony-bred, healthy adult fertile male Wistar rats (Rattus norvegicus) weighing between 150-200gm about 50-60 days aged were used for the present study. The animals were kept in polypropylene cages, measuring 430×270×150 mm and housed under controlled environment conditions with provision of 12hrs light: 12hrs dark conditions. The animals were fed with platted standard rat chow (Ashirwad, Chandigarh), supplemented with soaked gram and wheat, water was provided ad libitum. Body weight of each animal in all groups was measured weekly to see the possible changes in body weight throughout the experiment.

Experimental design and Protocol- Rats of similar body weight, size age were grouped as under. Whole study was divided into two experiments. Following experiments were carried out during the course of study, to observe antifertility effect and to observe mode of action/effects nature of the extract and reversibility effects.
The animals were divided into 5 treatment groups each consisting of 10 animals

Group-A: The animals were given sterile DW alone orally; serve as vehicle treated controls.
Group-B: The animals of this group were treated with Cassia tora (50%EtOH) at 50mg/kg.b.wt/day.
Group-C: The animals of this group were treated with Cassia tora (50%EtOH) at 100mg/kg.b.wt/day.
Group-D: The animals of this group were treated with of Cassia tora (50%EtOH) at 200mg/kg.b.wt/day.
Group-E: The animals of this group were treated with Cassia tora (50%EtOH) 100mg/kg. wt/day for 60 days were kept for a recovery period of 30 days.

Body organs weights- The initial final body weights of the animals were recorded. Testes, epididymis, seminal vesicles ventral prostrate were dissected out, freed from adherent tissues weighed to the nearest milligram on an electronic balance.

Tissue Biochemistry- The testis, epididymis, seminal vesicles ventral prostrate were dissected out, freed from adherent tissues weighted at nearest milligram balance. Protein [28], glycogen [29], cholesterol [30], sialic acid [31], ascorbic acid [32], fructose [33] were estimated in right side of testis other accessory reproductive organs.

Fertility Test- Male rats were introduced to female, 200-300 gm (male: female ratio, 1:2) at 17:00 h after 55 days of treatment. The mated females were allowed to complete the gestation. The number of pups was recorded litter size percent fertility was calculated [27].

Sperm Motility and Density- For sperm motility density, 50 mg of cauda epididymis was minced in 1 ml of physiological saline, immediately within 5 min after scarification; 1 drop of evenly mixed sample was applied to a glass slide under a cover glass. The percent motility was determined by counting both motile immotile spermatozoa per unit area. After that cauda epididymal sperm density was made by routine procedure express as millions/mm3 suspension [34].

Hormone Assay- Blood samples were also collected for serum separation to estimate FSH, LH and testosterone by radioimmunoassay. Serum samples separated by standard procedures stored at -20°C for subsequent analysis. Serum levels of testosterone were assayed in duplicate using radioimmunoassay kit [35].

Histopathological Study- Contra lateral side of the testis, epididymis, vas deferens, seminal vesicle, ventral prostate, kidney, heart liver were fixed in Bouin's fluid, dehydrated in graded ethanol, cleared in xylene and free from adherent tissue and embedded in paraffin wax (Melting point 55o-62oC). Sections were made at 6 µ was stained with Harris's hematoxylin and eosin to observe histopathological changes.

STATISTICAL ANALYSIS
Statistical analysis is based on biological statistics. All the values of body organ weights, biochemical estimations histometery were averaged expressed as Mean ± Standard error (S.E.). Data are expressed as mean ± S.E. analyze for statistical significance by using student's “t” test. The data considered as significant and highly significant at p<0.01 and p < 0.001, respectively [36].

Ethical Aspects- The study was carried out under the supervision of ethical committee of the Department of Zoology, University of Rajasthan, Jaipur [Vide Letter No.Rs/98/10/7454 dated 19/8/2010] and CPSEA [37] guidelines were followed to maintain the experimental animals.

RESULTS
Changes in blood and serum profile- Cassia tora extract treatment in rats caused a non significant change in serum serum proteins, Phospholipids, cholesterol, triglyceride, HDL, LDL and VLDL, acid phasphatase and alkaline phasphatase, LDH, SGOT, SGPT level after the treatment at different dose levels as compared to control rats. Cassia tora extract treated rats did not show any remarkable changes in blood and serum biochemistry and no significant alteration in hematological parameter (data not shown).

Effect on body and reproductive organ weight- Oral administration of extract of Cassia tora (fruit) for 60 days, caused no adverse effect on body weight; whereas the weight of testes and accessory sex organs decreased significantly (P<0.001) might be due to decreased level of androgens required to maintain the growth and development of reproductive organs. The weight reduction was dose dependent i.e. high dose 200mg/kg.b.wt. (Group D) treated group drastically reduced followed by less in low dose (Group C) 100mg/kg.b.wt. group. Significant changes were observed in vas deferens, seminal vesicle and ventral prostrate (Table 1).

Table 1: Effect on Body and organs weight of rats treated with Cassia tora extract

Treatment	Initial b.wt. (gm)	Final b.wt. (gm)	Testes mg/100gm b.wt.	Epididymides mg/100gm b.wt.	Vas deferens mg/100gm b.wt.	Seminal vesicle mg/100gm b.wt.	Ventral prostate mg/100gm b.wt.
Group A Control	120.00±2.35	156.50±1.67	1405.79±12.58	595.26±10.70	148.26±2.10	492.42±3.31	76.50±1.72
Group B Cassia tora 50 mg /kg.b.wt.	154.50±1.38	163.50±0.76	1399.51±18.63ns	574.76±9.93 ns	145.75±2.13ns	481.45±5.58 ns	73.28±0.95 ns
Group C Cassia tora 100mg/kg.b.wt	153.00±1.11	67.50 ±0.83	1381.06±5.86 ns	574.30±3.88 ns	142.83±2.30ns	479.26±7.80 ns	72.74±1.17 ns
Group D Cassia tora 200mg/kg.b.wt	114.00±2.08	166.50 ±0.76	1272.46±6.16**	472.74±6.26**	119.54±0.95**	427.80±3.02**	63.86±0.67**
Group E Cassia tora 100 mg / kg. b.wt. recovery	112.00±2.00	160.50±1.17	1374.63±9.70ns	581.88±8.36 ns	45.08±1.17ns	488.95±5.53 ns	75.81±1.17 ns

Data exposed as Mean ±S.E, ns = non-Significant,^* Significant (P=0.01),^** Highly Significant (P=0.001)

Biochemical changes- The ethanolic extract treatment of Cassia tora in rats were shown significantly decreased (P=0.001) level of protein, sialic acid, fructose, glycogen, cholesterol contents in reproductive organs. In recovery group (Group-E) after 30 days of withdrawal of treatment, it was altered up to non significant level (Table 2).

Table 2: Changes in level of protein, sialic acid, cholesterol, glycogen, ascorbic acid and fructose level in reproductive organs of rats following Cassia tora extract treatment

Treatment	Protein (mg/gm)		Sialic acid (mg/gm)		Cholesterol (mg/gm)	Glycogen (mg/gm)	Ascorbic acid (mg/gm)	Fructose (mg/gm)
	Testes	Cauda	Testes	Cauda	Testes	Testes	Adrenal	Seminal vesicle
Group A Control	226.19±3.41	223.08±3.23	5.20±0.25	5.44±0.41	16.06±0.47	2.17±0.08	2.90±0.22	4.89±0.31
Group B Cassia tora 50 mg/kg.b.wt	221.75±2.69 ns	219.53±3.52 ns	4.72±0.28ns	4.68±0.28ns	15.62±1.78 ns	2.16±0.06 ns	2.85±0.18ns	4.84±0.22 ns
Group C Cassia tora 100 mg / kg.b.wt	219.53±4.55 ns	216.42±3.63 ns	4.72±0.22 ns	4.60±0.14ns	15.69±0.48 ns	2.11±0.10 ns	2.81±0.12ns	4.36±0.23 ns
Group D Cassia tora 200 mg/kg.b.wt	186.65±3.86**	191.98±3.91**	4.08±0.23**	4.00±0.15*	14.12±0.63**	1.62±0.09**	2.43±0.16ns	3.61±0.26**
Group E recovery Cassia tora 100 mg / kg.b.wt	216.86±4.01 ns	219.09±5.05 ns	5.12±0.22 ns	4.60±0.23ns	15.69±0.66 ns	2.14±0.08 ns	2.80±0.18ns	4.81±0.20 ns

Data exposed as Mean ±S.E, ns = non-significant, ^* Significant (P=0.01), ^** Highly significant (P<0.001)

Effect on Sperm motility and density- Sperms motility and density in cauda epididymdes decreased significantly (P=0.001) in rats following extract treatment (Group B-D) in comparison to control rats (Group-A). Although, it was recover up to normal level in rats recovery (Group- E) (Fig. 1).

Fig. 1: Effetcts of Cassia tora extract treatment on sperm motility and density

Changes in hormone levels- The level of testosterone, LH and FSH analysis shows the decreased level of these hormones in Cassia tora extracts treated rats (Group B-D) in dose dependent manner at dose levels of 50 mg/ kg b.wt.,100 mg/ kg b.wt.,200 mg/ kg b.wt., however, in recovery group (Group- E) it return up to normal level (Fig. 2-4).\

      

Fig. 2: Effetcts of Cassia tora extract on testosterone level            Fig. 3: Effetcts of Cassia tora extract on LH level



Fig. 4: Effetcts of Cassia tora extract on FSH Level

Effect on spermatogenesis Photomicrographs of the testes of Cassia tora treated rats at the dose levels of 50 mg/ kg. b.wt., 100 mg/ kg. b.wt. and 200 mg/ kg. b.wt. caused degenerative changes. Germinal epithelium of seminiferous tubules was ruptured and wavy due to loosened connective tissue. Spermatogonia and sperm number were significantly reduced, cellular debris can be seen in lumen, inter tubular stroma was increased. However spermatogenesis alters up to normal level in rats of recovery group (Photomicrograph 1-5).



Photomicrograph-1, Group-A (X 100 H.E.) testis of control rats showing a normal histoarchitecture, well developed germinal epithelium, seminiferous tubules filled with spermatocytes, spermatids and spermatozoa
Photomicrograph-2, Group-B (X 100 H.E.) testes of rat treated at 50 mg/kg b.wt. showing degenerated epithelium, irregular and incomplete spermatogenesis
Photomicrograph-3, Group-C (X 100 H.E.) testes of rat treated at 100 mg/kg b.wt. showing decreased spermatocytes and sperms in seminiferous tubules
Photomicrograph-4, Group-D (X 100 H.E.) testes of rat treated at 200 mg/kg b.wt. showing incomplete spermatogenesis. Primary stages appear to be normal but later stages are absent
Photomicrograph-5, Group-E (X 100 H.E.) testes of rat treated with recovery showing normal semniferous tubule. Spermatocytes, spermatid and spermatozoa are clearly visible in the lumen. Sertoli cell and Leydig cell are also clearly seen

DISCUSSION
Maintenance of structure and functional integrity of accessory reproductive organs requires continuous supply of androgen [8,25, 38-41]. Oral administration of alcoholic extract of Cassia tora (fruit) for 60 days brought a decrease in the weight of reproductive organs indicating that the circulating level of androgen was not enough to maintain the weights of reproductive organs, however the weights of vital organs were not affected. [42-46].
Seminal vesicles are androgen-dependent and this property may be used as biological marker of androgen activity [47-49]. Protein level is directly correlated with the secretory activity of the epididymis, which in turn depends on the androgen levels [50-51]. The low levels of testicular protein are usually indicative of inhibition of spermatogenesis [52-54]. The decreased sialic acid level in testes and accessory organs of male rats and the epididymis can be a causative factor for impaired sperm function [39,55-56]. A decrease in glycogen content of the testis reduces the energy source for spermatogenic activity which could affect protein synthesis [42,57]. Decrease in ascorbic acid cause hypo functioning of testis and the degeneration of the germinal epithelium [58]. Fructose serves as source of energy for sperm, reduction in the fructose might be due to decreased secretory activity of seminal vesicle [59-60].
Sperm density and motility directly correlates with fertility chances and therefore decreased sperm density [50-54 and motility of spermatozoa might reduced fertility of rats followed extract treatment [61-70]. A large number of metaphasic cells in the germ epithelium of treated animals might be caused by cell cycle blockage or [46,71] arrest at the spermatid level in the form of degenerative changes in the germinal cells together with few fragmented sperms in the lumen and acquired a thick, irregular basement membrane [72-74].
Spermatogenesis requires LH and FSH for initiation and maintenance in male rats. LH, through specific receptors found on the surface of Leydig cells, controls the production and secretion of testosterone. Normal testicular function is dependent on FSH and testosterone is absolutely required for normal spermatogenesis [75-80]. FSH establishes a quantitatively normal Sertoli cell population, whereas androgen initiates and maintains sperm production, thus both hormones co-operate via independent functions to enable maximal spermatogenic output [81-85].

CONCLUSIONS
It can concluded that treatment of Cassia tora fruits extract in male rats reduced levels of protein, fructose, ascorbic acid, glycogen, and sialic acid contents might be responsible to caused degenerative changes in germinal epithelium of seminiferous tubules of testes. The decreased level of testosterone further support anti-androgenic effects of the treatment resulted decreased sperm density and motility reduced fertility of extract treated rats.places.

ACKNOWLEDGMENT
Authors are grateful to The Head and Coordinator, Centre for Advanced Studies, Department of Zoology, University of Rajasthan, Jaipur for laboratory facilities, State DST Govt. of Raj, Jaipur, and UGC, New Delhi for partial financial support.

REFERENCES

1. Mali PC. Control of fertility in male Wistar rats treated with hydro alcoholic extract of Withania somnifera fruits. Int J Pharm Bio Sci, 2013; 5: 13-21.
2. Sharma D K and Mali PC. Inhibition of Spermatogenesis with the Treatment of 50% Methanolic extract of Maytenus emarginata leaves in Albino Rats. Int J Pharm Sci Res, 2016; 7(8): 3379-87.
3. Chowdhury SR, Gupta CM, and Kamboj VP. Current status in fertility regulation: indigenous and modern approaches. Asn J Androl, 2001:99-119.
4. Mali PC, Ansari AS, and Chaturvedi M. Effects of chronically administered Martynia annua root extract on male rats. J Ethnopharmacology,2002; 82 (2-3): 61- 67.
5. Sharma DK, Luhadia G, Soni PK, and Mali PC. Traditionally used Indian medicinal plants exhibits contraceptive activities: A Review. Int J Pharmacol Bio Sci, 2015; 9:39-45.
6. Gupta RS, Kanwar M, Rehwani H, Verma SK, and Dobhaal MP. Contraceptive efficacy of Strychnos potatorum seed extract in male albino rats. Asn J Exp Sci, 2006; 20(1): 181-187.
7. Luhadia G, Sharma DK, Soni PK, and Mali PC. Exploration of traditional medicinal plants for antifertility effects: A Review. Adv Pharmacol Toxicol, 2015; 16: 65-71.
8. Soni, PK, and Mali PC.Oral administration of petroleum ether extract of Tecomella undulata leaves affects spermatogenesis and fertility of male rats. Europ J Biomed Pharmaceut Sci, 2016; 3: 339-344.
9. Kumar S,Dagar S,Kumar P,Singh J,Kumar S, and kumar D. Antifertility effect of hydroalcoholic extract of Pandanus odoratissimus L. leaves. Porto Biomed J, 2017; 2(5): 167-169.
10. Dahanukar SA, Kulkarni RA, and Rege NN. Pharmacology of medicinal plants and natural products. Ind J Pharmacol, 2000; 32: 81-118.
11. Chaudhary R, Ranjan A, and Mali PC.Reversible contraceptive efficacy and safety evaluation of ethanolic extract of Maytenus emargineta in male albino rats. J Pharmacy Research, 2011; 4(1): 213-216.
12. Stone BC. The flora of guam. A Manual for the identification of the vascular plants of the Island. Micronesica, 1970; 6 (1/2): 1-659
13. Wong SM, Wong MM, Seligmann O, and Wagner H. New antihepatotoxic naphtha-pyrone glycosides from the seeds of Cassia tora. Planta Med, 1989; 55:276-280.
14. Zhang Z, and Yu B. Total synthesis of the antiallergic naphtha-a-pyrone tetraglucoside, Cassia side C2, isolated from Cassia seeds. J Org Chem, 2003; 68:6309-6313.
15. Choi JS, Lee HJ, Park KY, Jung GO, and Kang SS. In vitro antimutagenic effects of naphthopyrone glucosides from Cassia tora. Planta Med, 1998; 64:100-104.
16. Kim YM, Lee CH, Kim HG, and Lee HS. Anthraquinones isolated from Cassia tora (Leguminosae) seed show an antifungal property against phytopathogenic fungi. J Agric food Chem, 2004; 52:6096-7000.
17. Ladell C, Mc Donald GM, and Friedman M. Composition of sticklepod (Cassia obtusifolia ) toxic weeds seeds. J Agri Food Chem, 1990; 38:2169-2175.
18. Choi JS, Lee HJ, and Kang SS. Alaternin, Cassia side and rubrofusarian gentiobioside, radical scavenging principles from the seeds of Cassia tora on 1,1-diphenl-2-prcrylhydrazyl (DPPH) radical. Arch Pharm Res, 1994; 17:462-466.
19. Hatano T, Mizuta S, Ito H, and Yoshida T. C-glcosidic flavonoids from Cassia occidentalis. Pytochemistry, 1999; 52(7):1379-1383.
20. Maity TK and Dinda SC. Purgative activity of Cassia tora leaf extract and isolated aloe-emodin. Int J Pharmaceu Sci, 2003; 65(1):93-95.
21. Patil UK, Saraf S, and Dixit VK. Hypolipidemic activity of seeds of Cassia tora (Linn.). J Ethnoparmacol, 2004; 90:249-252.
22. Zhenbaoa J, Feia T, Lingb G, Guanjuna T, and Xiaolin D. Antioxidant properties of extracts from juemingzi Cassia tora (Linn.) evaluated in vitro. Life Sci, 2004; 76(1): 1985-2001.
23. Shrivastava AK. Medicinal and herbal species, medicinal plants, APH Publishing corporation New Delhi, 2006;121:52-53.
24. EL Halawany AM, Chung MH, Nakamura N, Ma CM, Nishihara T, and Hattori M. Estrogenic and anti-estrogenic activities of Cassia tora phenolic constituents. Chem Pharm Bull, 2007; 55(10):1476-1482.
25. Ghogar A, and Jiraungkoorskul W. Antifertility effect of Bougainvillea spectabilis or Paper Flower. Pharmacogn Rev, 2017; 11(21): 19–22.
26. Sala AV. Indian medicinal plants, Orient Longman Pvt. Ltd. New Delhi, 2002; Vol. II and V: 11, 19, 26: 409-411.
27. WHO. Annual technical Report 1998 Special programme of Research Development and Research Training in Human reproduction. World Health Organization, Geneva 1983.
28. Lowry OH, Rosenbrough NJ and Far AL. Protein measurements with the folin phenol reagents. J Biol Chem, 1951; 193: 265-275.
29. Montgomery R. Determination of glycogen. Arch Biochem Biophys, 1957; 73: 378-389.
30. Mann T. The biochemistry of semen and the male reproductive tract Mc Theun, London, 1964:239.
31. Warren L. Warren L. The thiobarbituric acid assay of sialic acid. J Biochem, 1959; 8: 1971-1975.
32. Roe JH and Kuther CA. Detection of ascorbic acid in whole blood and urine through the 2, 4, DNPH derivative of dehydroascorbic acid. J Biochem, 1943; 147: 399-401.
33. Foreman D. A modification of the Roe procedure for determination of fructose in tissues with increase specificity. Analyt Biochem, 1973; 56: 584-590.
34. WHO Protocol MB-50: A method for examining the effect of the plant extracts administration orally on the fertility of male rats (APF/ IP, 99914E) Geneva World Health Organization 1983.
35. WHO. Annual technical Report 1998 Special programme of Research Development and Research Training in Human reproduction, world Health Organization, Geneva 1999.
36. Gupta S. Sampling and test of significance In: Gupta S (eds.) Statistical Methods. Sultan chand and sons Publishers, New Delhi, 1978:58-76.
37. CPCSEA. Committee for the Purpose of Control and Supervision on Experiments on animals, New Delhi, 2006: Indian Council of Medical Research.
38. Gupta RS, Kachhawa JB, and Chaudhary R. Antifertility effects of methanolic pod extract of Albizzia lebbeck. Asian J Androl, 2004; 6: 155–159.
39. Mohan VR, Shajeela PS, Jesudas LL, and Tresina SP. Antifertility activity of ethanol extract of Dioscorea esculenta (Linn.) Schott on male albino rats. Int J Pharm Tech Res, 2011; 3(2):946-954.
40. Yakubu MT. Effect of a 60-day oral gavage of a crude alkaloid extracts from Chromolaena odoratum leaves on hormonal and spermatogenic indices of male rats. J Androl, 2012; 33(6):1199-1207.
41. Mali PC, Singh AR and Verma MK. Contraceptive effects of Withanolide- A in adult male albino rats). Adv Pharmacol Toxicol, 2015; 16: 31-44.
42. Bone W, Jones NG, Kamp G, Yeung CH, and Cooper TG. Effect of ornidazole on fertility of male rats. Inhibition of glycolysis and related motility pattern and zona binding required for fertilization in vitro. J Rep and Fertil, 2000; 118:127-135.
43. Kamischke A, Heuermann T, Kruger K, Von Eckardstein S, Schellschmidt I, Rubig A, and Nieschlag E (An effective hormonal male contraceptive using testosterone undecanoate with oral or injectable norethisterone preparations. J Clin Endocrinol Metab, 2002; 87: 530-539.
44. Venkatesh V,Sharma JD, and Kamal R. A comparative study of effect of alcoholic extracts of Sapindus emarginatus, Terminalia belerica, Cuminum cyminum and Allium cepa on reproductive organs of male reproductive organs of male albino rats. Asn J Exp Sci, 2002; 16(1&2):51-63.
45. Gupta RS, Bhatnager AK, Joshi YC, Sharma MC, Khushalani V, and Kachhawa JB. Induction of antifertility with lupeol acetate in male albino rats. Pharmacol, 2005; 75(2):57-62.
46. Raji Y, Akinsomisoye O, and Salman TM. Antispermatogenic activity of Morinda lucida extract in male rats. Asn J Androl, 2005; 7(4):405-410.
47. Gonzales G, and Villena A. True corrected seminal fructose level:a better marker of the function of seminal vesicles in infertile men. Int J Androl 24, 2001; (5): 255-260.
48. Barlow NJ and Foster PM. Pathogenesis of male reproductive tract lesions from gestation through adulthood following in utero exposure to Di(n-butyl) phthalate. Toxicol Pathol, 2003;31(4):397–410.
49. Barlow NJ, Mcintyre BS, and Foster PMD. Male reproductive tract lesions at 6, 12, and 18 months of age following in utero exposure to di(n-butyl) phthalate. Toxicol Pathol, 2004; 32:79-90.
50. Aladakatti RH, Suresh B, Jadaramkunti UC, and Hiremath MB. Effect of graded doses of nimbolide (A major componemt of Azadirachta indica leaves) on biochemical and sperm functional parameters in male albino rats. J Laborat Anim Sci, 2011; 1:24-30.
51. Ghodesawar MG, Usharani G, and Aladakatti RH. Carbohydrate nature of lyophilized leaf powder of Azadirachta indica induced inhibition of the activities of accessory reproductive ducts in male rats. Int Multidiscipli Res J, 2012; 2(10):17-26.
52. Vijaykumar B, Sangamma I, Sharanabasappa A, and Patil SB. Antispermatogenic and hormonal effects of Crotalaria juncea (Linn.) seed extracts in male mice. Asn J Androl, 2004; 6(1):67-70.
53. Chinoy NJ, Dilip T, and Harsha J. Effect of Carica papaya seed extract on female rat ovaries and uteri. Phytother. Res, 2006; 9(3): 169-165.
54. Thejashwini MS, Krishna RH, and Shivabasavaiah. Reversible antifertility effect of cyamposis psoralioides in male swiss albino mice. Int J Adv Biol Res, 2012; 2(3):416-424.
55. D'cruz SC and Mathur PP. Effect of piperine on the epididymis of adult male rats. Asn J Androl, 2005; 7(4):363-368.
56. Gupta RS, Kachhawa JBS, and Sharma A. Effect of methanolic extract of Dendrophthoe falcata stem on reproductive function of male albino rats. J Herb Pharmacother, 2007; 7(2):1-13.
57. Sethi J, Yadav M, Sood S, Dahiya K, and Singh V. Effect of tulsi (Ocimum Sanctum Linn .) on sperm count and reproductive hormones in male albino rabbits. Int J Ayurveda Res, 2010; 1(4):208-210.
58. Goyal S, Manivannan B, Ansari AS, Lohiya NK. Safety evaluation of long term treatment of methanol sub-fraction of seeds of Carica papaya as a male contraceptive with particular emphasis on carcinogenicity in albino rats. Int J Pharmacol. 2009; 5:114–25
59. Singh SK and Mishra RK. Antispermatogenic and antifertility effects of fruits of Piper nigrum (Linn.) in mice. Ind J Exp Bio, 2009; 47(9):706-714.
60. Valentine U. Antifertility effects of aqueous extract of Phyllanthus niruri in male albino rats. Int. Sci. Pub., 2011, 4(2), 1-14.
61. Sarkar M,Gangopadhyay P,Basak B,Chakrabarty K,Banerji J,Adhikary P, and Chatterjee A. The reversible antifertility effect of Piper betle (Linn.) on swiss albino male mice. Contraception, 2000; 62(5):271-274.
62. Ghosesawar MG, Ahamed NR, Ahmed M, and Aladdkatti RH. Azadirachata indica adversely affects sperm parameters and fructose levels in vas deferens fluid of albino rats. Basic and Clin Physiol and Pharmacol, 2003:387-395.
63. Patil SB, Seetharam YN, Sujeeth H, Jyothishwaran G, Barad A, Sharanabasappa G, Umareddy B, and Vijaykumar MB. Antifertility effect of ethanolic extract of Amalakyadi churna in male albino mice. Asn J Androl, 2003; 5:247-250.
64. Noarkes DE, Parkinson TJ, England GCW, and Arthur GH. Normal reproduction in male animals. In: Arthur’s Veterinary Reproduction and Obstetrics. 8th ed. Saunders Publishers, Edinburgh, 2004:673-694.
65. Liu YD and Baker HWG. High frequency of defective sperm–zona pellucida interaction in oligozoospermic infertile men. Hum Rep, 2004; 19(2):228-233.
66. Choi EM and Hwang JK. Investigations of anti-inflammatory and antinociceptive activities of Piper cubeba, Physalis angulate and Rosa hybrid. J Ethnopharmacol, 2003; 89:171 – 175.
67. Fukushima M, Koh E, Choi J, Maeda Y, Namiki M, and Yoshida, A. Reevaluation of azoospermic factor c microdeletions using sequence-tagged site markers with confirmed physical positions from the GenBank database. Fertil Steril. 2006; 85: 965–97168.
68. Yinusa R, Akinsomisoye O S, Salman T M. Antispermatogenic activity of Morinda lucida extract in male rats.Asian J Androl, 2005; 7:405-410.
69. Saba AB, Oridupa OA, Oyeyemi MO, and Osanyigbe OD. Spermatozoa morphology and characteristics of male wistar rats administered with ethanolic extract of Lagenaria Breviflora Roberts. Afr J Biotechnol, 2009; 8(7):1170-1175.
70. Kumar BS, Sathiyaraj K,Sivaraj A, Thirumalai T, Baskaran N, Vinothrasu K, and Inbasekar P. Antifertility activity of aqueous leaf extract of Andrographis paniculata in male albino rats. Int J Pharmaceut & Biol Arch, 2011; 2(4):1179-1182.
71. Kushwaha S, Agarwal M, Mutreja A, and Chauhan A. Impact of 50% ethanolic extract of Calendula officinalis (flower) on the reproductive function of male albino rats. Egtn J Biol, 2007; 9:42-46.
72. Khattab FKI. Histological and ultrastructural studies on the testis of rat after treatment with aluminium chloride. Autn J Basic and Applied Sci, 2007; 1(1):63-72.
73. Ahmed M, Ahmed RN, Aladakatti RH, and Deepthi KR. Reversible histoarchitecture study of testis and cauda epididymis and changes in cauda epididymal epithelial cell types on treatment with benzene extract of Ocimum sanctum leaves in albino rats. Oriental Pharm Exp Med, 2008; 8(2):111-124.
74. Singh A and Singh SK. Reversible antifertility effect of aqueous leaf extract of Allamanda cathartica (Linn.) in male laboratory mice. Andrologia, 2008; 40(6):337-345.
75. O'Donnell L, Robertson KM, Jones ME, and Simpson ER. Estrogen and spermatogenesis. Endocrinol Rev, 2001; 22:289-318.
76. McLachlan RI, O'Donnell L, Meachem SJ, Stanton PG, De Kretser DM, Pratis K, and Robertson DM. Identification of specific sites of hormonal regulation in spermatogenesis in rats, monkeys, and man. Recent Prog Horm Res, 2002; 57:149-179.
77. Haywood M, Spaliviero J, Jimemez M, King NJ, Handelsman DJ, and Allan CM. Sertoli and germ cell development in hypogonadal (hpg) mice expressing transgenic follicle-stimulating hormone alone or in combination with testosterone. Endocrinol, 2003; 144: 509-517.
78. Khouri NA and EL-Akawi Z. Antiandrogenic activity of Ruta graveolens (Linn.) in male albino rats with emphasis on sexual and aggressive behaviour. Neuroendo letters, 2005; 26(6):823-829.
79. Matthiesson KL, McLachlan RI, O'Donnell L, Frydenberg M, Robertson DM, Stanton PG, and Meachem SJ. The relative roles of follicle-stimulating hormone and luteinizing hormone in maintaining spermatogonial maturation and spermiation in normal men. J Clin Endocrinol Metab, 2006; 91:3962-3969.
80. O'Donnell L, Meachem SJ, Stanton PG, and McLachlan RI. Endocrine regulation of spermatogenesis. In: Knobil and Neill's physiology of reproduction, ed Neill JD Elsevier, San Diego, CA, 3rd ed 2006:1017-1069.
81. Chaturvedi M, Mali, PC and Dixit VP. Antifertility effects of the roots of Echinops echinatus (Roxb.) in male rats J Environ and Pollu 1995;2 (4): 153- 157
82. Morakinyo A, Adeniyi O, and Arikawe A. Effects of Zingiber officinale on reproductive functions in the male rat. Afr. J. Biomed. Res, 2008; 11: 329-34.
83. Handelsman DJ. Hormonal regulation of spermatogenesis: insights from constructing genetic models. Rep Fertil Dev, 2011; 23:507-519.
84. Onyeka AC, Aligwekwe AU, Olawuyi TS, Nwakanma AA, Kalu EC, and Oyeyemi AW. Antifertility effects of ethanolic root bark extract of Chrysophyllum albidum in male albino rats. Int J Appl Res Natur Prod, 2012; 5(1): 12-17.
85. Abel MH, Baker PJ, Charlton HM, Monteiro A, Verhoeven G, De Gendt K, Guillou F, and O'Shaughnessy PJ. Spermatogenesis and Sertoli cell activity in mice lacking Sertoli cell receptors for follicle-stimulating hormone and androgen. Endocrinol, 2008; 149:3279-3285.

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How to cite this article: Khan S, Mali PC: Reversible Antifertility Effect of Cassia tora Linn in Male Rats. Int. J. Life. Sci. Scienti. Res., 2017; 3(5): 1415-1423. DOI:10.21276/ijlssr.2017.3.5.26 Source of Financial Support:Nil, Conflict of interest: Nil