Effect of Y virilin on male infertility.NN Rege, J Date, V Kulkarni, AR Prem, SV Punekar, SA Dahanukar
Department of Pharmacology, Seth G.S. Medical College, Parel, Mumbai, India., India
A prospective placebo controlled double blind study was conducted in patients attending male infertility clinics of our hospital to evaluate effects of a herbal formulation for male infertility--'Y-virilin'. In phase 1 forty patients with oligospermia with or without asthenospermia were randomly allocated to 2 treatment groups--Treatment Group A i.e. formulation under test and treatment Group B (Placebo). Therapy with these agents was given twice a day for 6 months. In phase 2, 12 patients with azospermia were administered either 'Y virilin' or the placebo (n = 6/Gp). In all patients along with semen analysis (sperm count, percentage of motile sperms and grade of motility) was done monthly for 6 months. Serum FSH levels were estimated before and at the end of therapy. A significant increase in sperm count was observed from 2-3 months in oligospermics receiving Y virilin as compared to basal values (p < 0.05). In Group B the follow-up sperm counts were either comparable to basal values or were lesser. However, the percentage and grade of motility did not differ in two groups at the end of respective treatment. No change was found in mean FSH value. During the therapy period incidence of conception was 20% in treatment Group A and 5% in Group B. Of the azospermic receiving 'Y-virilin' 50% showed a count of 10-20 millions/cmm while none from the placebo group. This study highlights the therapeutic potential of the tested formulation in the patients with infertility.
Keywords: Double-Blind Method, Follicle Stimulating Hormone, blood,Human, Male, Oligospermia, blood,drug therapy,Phytotherapy, Prospective Studies, Sperm Count, drug effects,
In 50% of infertile couples, the male factor is at least partly responsible for failure to conceive. In the recent years attention has been focussed on the evaluation of male partner and tests apart from semen analysis are being developed to identify the precise cause for infertility. However, success of various therapeutic regimens remains low.
In fact, since time immemorial, scarcity of drugs for male infertility has been the concern for physicians. A look at our ancient Ayurvedic literature reveals that the subject of 'Infertility and agents/methods promoting fertility' constitute a separate branch in this science. Various agents mentioned therein for male infertility and impotency are classified as a. Vrishya (those increase potency,) b. Vajikar (those impart sexual pleasure,,) and c. Shukral (those increase sperm counts and improve properties of seminal fluid).
Many formulations containing agents from these categories flood the Indian market even today. However, there is no scientific study carried out to validate their efficacy.
‘Y virilln' is one such formulation (marketed by Dr. Paranjape, Akshaya Pharmaceuticals) prescribed for male infertility and contains the ingredients shown in [Table:l].
It was decided to evaluate the claims put forth for this formulation by conducting a prospective clinical trial in male patients with infertility.
Initially the protocol was planned for patients with oligospermia (Phase 1). After analyzing the results of this phase it was extended to patients with azospermia (Phase 2).
After receiving an approval from the Ethics Committee of our Institution, a randomized double blind study was conducted on male patients attending Infertility Clinic. The selection criteria were patients with evidence of oligospermia with/without associated asthenospermia and showing no response to the standard modern medicinal therapy as judged from 3 consecutive sperm counts done at monthly interval.
Patients with azospermia, congenital testicular deformity, absent vas or those with surgically correctable obstructive lesions were excluded from the study.
After obtaining written informed consent the selected 40 patients were randomly allocated to two treatment groups - Group A and Group B. Patients (n=20) from group A were given Y-virilin in a dose of 1 capsule twice a day for 6 months. Patients from Group B received equivalent dose of placebo (n=20). Patients from both the groups were followed up monthly for a period of 6 months.
Before starting the therapy and at each follow-up semen analysis was carried out. A record of sperm count, percentage of motile sperms and grade of motility was maintained for each examination. Motility index was determined by multiplying percentage of motile sperms and grade. Assays for serum FSH were carried out before and after 6 months of therapy. This parameter was included as a probe to study effect of drug on hypothalamo-pituitary axis.
The data were analyzed statistically at the end of the study following which decoding was carried out. Paired 't' test was used to compare the change in sperm count. Changes in percentage of motile sperms and grade of motility were analyzed using nonparametric tests.
Twelve patients with azospermia were selected for the study. Testicular biopsy done of 11/12 patients revealed thickening of basement membrane (n=6), maturation arrest (n=4) and germ cell hypoplasia (n=l). The patients were randomly allocated to Group A (Y virilin) And Group B (placebo) (distribution is shown in [Table - 2]) and followed-up monthly for 6 months for changes in sperm counts.
The basal counts for patients in Group A and Group B were comparable [22.9 + 14.5 (range: 6-55) millions/cmm and 20.7 + 17.67 (range : 5-50) millions/cmm for Group A and B respectively].
Number of spermatozoa counted at each examination was compared with basal sperm count for each patient. The effects of 'Y virilln' and placebo on sperm count are depicted in [Figure - 1]. A significant increase in count was observed following one month therapy with 'Y virilin'. The maximum increase in count was reached after 2 or 3 months of treatment. Thereafter the counts either remained static or started decreasing. However they were significantly higher than the basal values (p < 0.05). On the other hand, in patients receiving placebo (Group B) no change in counts was observed at monthly examination as compared to basal values for first 4 months. The counts in last 2 months were significantly lower than basal values (p < 0.05).
The semen analysis of patients from Group A showed that grade of motility ranged from I-II and 10-50% sperms were motile. In 11/20 patients motility index was 60 or lesser (Normal: 80-240). The patients from Group B also exhibited same degree of motility (Grade I-II) and percentage of motile sperm was also the same at the time of inclusion (10-50%). In 12/20 patients motility index was 60 or less.
No significant change in either the grade of motility or percentage of motile sperms was observed following therapy with drug or placebo.
Serum FSH levels
Both the treatment groups were comparable with respect to the serum FSH levels at the time of inclusion (Group A : 15.04 + 5.41 mIU/ml and Group B : 15.6 + 9.6 mIU/ml). No change in mean FSH was observed after 6 months of therapy (Group A : 11.4 + 5.5 mlU/ml and group B: 13.49+ 7.75 IU).
Conception incidence was 20% for patients receiving ‘Y virilin’ (4/20). In the group B. the incidence was 5% (1/20).
In 3 of 6 azospermic patients taking 'Y-virilin' counts increased to 12-20 millions/cmm at the end of therapy [Figure - 2]. The basal testicular biopsy of these patients showed maturation arrest. On the other hand 5 of 6 patients receiving placebo remained azospermic and in one patient a count of 2-3 sperms was reported. The patients exhibiting rise in sperm counts did not report for repeat testicular biopsy. The basal FSH levels at the time of inclusion were 10.25 + 2.8 mIU/'ml in Group A and 10.6 + 6.19 mlU/ml in placebo group. No change in mean FSH was observed after 6 months of therapy in both the groups (8.38 + 2.36 mIU/ml and 7.85 + 7.8 mIU/ml in Group A and B respectively).
'Y virilin' in a given dose produced significant increase in sperm counts in all the patients with oligospermia with or without asthenospermia. The point to note is all these patients were non-responders to the modern medicinal therapy.
Improvement in sperm counts in oligospermic patients can increase chances of pregnancy. For any drug claimed to have pro-fertility effect, the end point should be incidence of conception. On this basis, a group receiving 'Y virilin' shows a trend towards increase in incidence of conception.
In spermatogenesis 74 days are required for spermiation and 10-14 days for epididymal passage of sperms. In this study a significant rise in sperm count was found after 2-3 months of therapy which indicates possible effect of drug on the spermatogenic process.
No change was found in serum FSH values following treatment with 'Y-virilin' which indicates that the rise in sperm counts induced by this agent was not mediated through modulation of hypothalamo - pituitary axis. This also suggests the possibility that the effect of ‘Y virilin’ may be due to the direct effect on the germinal epithelium.
To test this possibility the effect of drug was evaluated in the patients with azospermia. As seen from the results, in 50 % patients with azospermia the counts have increased. All these patients had maturation arrest. It therefore appears that the site of action may be the germinal epithelium where it influences spermatogenesis and maturation. Unfortunately, these patients did not report for post-treatment biopsy. Therefore, any direct evidence for such effect is lacking.
Nevertheless the efficacy of Y virilin in non-responding oligospermics and to some extent in azospermic have raised a new hope in the light of non-availability of effective agent in modern medicine. Further research is necessary to find out which of the plant/s in formulation is responsible for the effect and also the mechanism of its action.
[Figure - 1], [Figure - 2][Table - 1], [Table - 2]