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Male antifertility activity of Azadirachta Indica in mice.
Azadirachta Indica A Juss (Nimba) is a plant that grows throughout the greater part of India and Burma.' The medicinal properties of Azadirachta Indica have been described in Ayurveda, by Sushruta as well as in ancient books like Rasarathasamucchaya, Sarangadhara, Bhavaprakasha and Bhisagya Ratnavali etc. The bark and leaves are used as bitter tonic and astringent. It is considered as an antiseptic, blood purifier and useful in skin disorders. The fruit is used as emolient and purgative. Azadirachta Indica is also considered beneficial in the treatment of a wide range of disorders like cough, nausea, vomiting, fever, jaundice, gonorrhoea, urinary tract infection, intestinal worm infestation and leprosy.[1], [5] The juice of fresh green leaves of Azadirachta Indica is believed to suppress 'Kam Vasana'. It was consumed by sanyasees in shrines and the pupils studying in Gurukul for the same purpose. The antifertility activity of Azadirachta Indica has been mentioned previously.[2] ,[4], [6] The anti-ovulatory activity is studied by the contraceptive testing unit of Indian Council of Medical Research.[2] The results are negative. There is no documented evidence referring to the male antifertility activity of Azadirachta Indica. It was therefore of interest to investigate the male antifertility activity of the plant.
Adult healthy male and female mice weighing between 25 and 30 gms were included in the study. Twenty five mice were divided into two groups. The control group containing 10 animals received normal saline in the dose of 10 ml/kg., orally every day for one month. The Azadirachta Indica group containing 15 animals received freshly prepared, water extract of crushed green leaves of Azadirachta Indica in the dose of 1 ml per mouse orally every day for one month. Fifty grammes of fresh green leaves were crushed in the homogeniser after addition of 100 ml of glass-distilled water. The juice thus obtained was filtered through a gauze piece and used for oral feeding. At the end of one month, five animals from each group were sacrificed and the testes were subjected to the histological examination. The rest of the animals were mated with adult healthy nonpregnant females. Prior to mating the females were isolated for one month to rule out pre-existing pregnancy. Three females and one male were placed in one cage. The females were examined every day. The presence of vaginal plug was taken as day one of pregnancy. The number of pregnant females and litter size was noted in the control and Azadirachta Indica treated groups. The males treated with Azadirachta Indica were separated after one month. Fifteen days after this i.e. after a total drug-free interval of 12 month, the animals were again mated with healthy, adult, nonpregnant females and the number of pregnancies and the litter size was noted. Total number of animals used in the study were 88, 25 being males and 63, females.
These are presented in [Table 1] , [Table 2], [Table 3] The weights of the animals in control and Azadirachta Indica group were comparable before as well as after the treatment [Table 3]. It was observed that control mice showed 100%, fertility rate. In Azadirachta, Indica treated animals the number of pregnancies as well as the litter size was reduced. After one and half month of drug free interval, the male mice showed a normal reproductive function as indicated by the number of pregnancies. Histological examination of testes revealed no evidence of inhibition of spermatogenesis.
It can be seen from the results that Azadirachta Indica possesses reversible male antifertility activity. The antifertility effect can be produced in the male by interference at the following steps: (1) Secretion and release of FSHRF and LHRF from the hypothalamus. (2) Secretion and release of FSH and LH from the pituitary. (3) Production of spermatozoa and androgenic hormone. (4) Transport of sperms into the epididymis with simultaneous maturation of spermatozoa. (5) Passage of spermatozoa through vas deferens to ampulla. (6) Suspension of sperms in seminal plasma during ejaculation. (7) Passage of ejaculate through cervix. (8) Ascent of sperms through the uterus and oviducts. (9) Penetration of ovum by spermatozoa. Spermatogenesis and functional maturation are under the influence of pituitary gonadotrophins. Evidence is now available that both FSH and LH are required for the development and maintenance of testicular function.[3] The histological picture shows that Azadirachta Indica has no effect on the spermatogenesis. Infertility can occur without histological changes with the sperms remaining fully motile. This is designated as functional sterility.[8] Azadirachta Indica therefore is thought to have the effect on the steps 4, 5, 6, 7, 8 and 9 of reproduction. This is a preliminary report on the antifertility activity of Azadirachta Indica in male mice, which indicates that the antifertility activity is not due to inhibition of spermatogenesis. However, extensive detailed studies on large number of animals and of different species will only confirm this observation.
We thank Dr. V. R. Deshpande, Dean, Medical College, Aurangabad for allowing us to carry out this work. Kind help given by the Heads of the Departments of Anatomy and Pathology is appreciated.
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