Assessment of a Polyherbal Ayurvedic Medicine for Sexual Activity in Rats

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1 [Indian Drugs (1999): (36), 9, ] Assessment of a Polyherbal Ayurvedic Medicine for Sexual Activity in Rats Kuram Ali Ahmed and Venkataraman, B.V., Department of Pharmacology, St. John s Medical College, Bangalore, India and Mitra, S.K.*, R&D Centre, The Himalaya Drug Company, Bangalore, India. [*Corresponding Author] ABSTRACT A polyherbal Ayurvedic Medicine (PAM-F) is widely used for treating male sexual dysfunction in the Indian subcontinent. In this animal model study, evaluation of acute and chronic effect of a newly formulated herbal product, PAM-R was assessed in sexually normal and sluggish male rats compared with PAM-F (Tentex forte). The preparation showed beneficial effects of various degrees with respect to the sexual behaviour of rats. The mean sperm count was also higher in the drug treated animals. Among the two herbal preparations, PAM-R (Tentex Royal) produces a more sustained increase of sexual activity. INTRODUCTION The search for drugs improving sexual behaviour dates back to ancient ages. Sexual behaviour is hormone dependent 1. Hence, sex hormones are popularly prescribed for enhancing sexual vigour. Hormones may control sexual activity by altering the monoamine transmitter activity 2-8. Modern medicine does not possess authentic drugs for improving the sexual function. Herbal preparations containing Orchis latifolia, Alpinia galanga, Mucuna pruriens, are used to improve sexual behaviour and to possess androgenic and anabolic actions. PAM-F is an Ayurvedic preparation useful in impotence in men 9. Each 250 mg of PAM-F contains: Hibiscus abelmoschus 8 mg, Withania somnifera 50 mg, Argyreia speciosa 24 mg, Mucuna pruriens 24 mg, Trivang 24 mg, Shilajeet 24 mg, Crocus sativus 15 mg, Strychnos Nux vomica (detoxified) 12 mg, Makardhwaj 12 mg, Orchis mascula 12 mg, Anacyclus pyrethrum 12 mg, Sida cordifolia 12 mg, Bombax malabaricum 12 mg and Piper nigrum 3 mg. Recently a new polyherbal formulation called PAM-R has been added to this list of sex tonics. Each 250 mg of PAM-R contains: Anacyclus pyrethrum 90 mg, Asparagus adscendens 90 mg, Aconitum napellus 25 mg and Piper betle 25 mg. PAM-R contains only 4 ingredients of which Asparagus adscendens and Aconitum napellus are not present in the PAM-F. PAM-R is also found to be useful for reversing the deleterious effects of alcohol on the male reproductive system in rats 10. Therefore, in the present study PAM-R was tested for the effects on various aspects of sexual function and compared with PAM-F. MATERIALS AND METHODS Drugs All the drugs were polyherbal formulations derived from Ayurveda and herbs in each drug were procured from authentic sources and were identified by botanists. PAM-F, an established polyherbal drug was taken as a standard for PAM-R. Diluting medium for sperm count was prepared by dissolving 50 g of sodium bicarbonate in 10 ml of 40% formalin.

2 After adding 5 ml of a saturated aqueous solution of Gentian violet, final volume of 1000 ml was made up with distilled water. Estrogen (german Remedies, India), progesterone (German Remedies, India) and chemicals of analar grade were used. Animals Wistar male and female rats of 3 months old were used for acute experiments, whereas for the chronic studies one month old rats were selected. All the rats were procured from Indian Institute of Science, Bangalore. Female rats were housed in groups of two and males were housed singly in polypropylene cages with free access to standard feed [Brook Bond Lipton (India) Ltd.] and tap water. A reversed twelve hour light-dark cycle was employed with fluorescent ceiling light. A dim red light was provided during dark cycle 11. All the rats were allowed 2 weeks to adjust to the environment prior to the experiment. Preparation of females Female albino rats were overectomized at 80 days of age and allowed 10 days to recover from surgery. After ovariectomy, the animals were in diestrous stage. To bring them to estrous, they were given 2 µg/mkg oestrogen orally 48 h before test and 500 µg/kg of progesterone 4-6 h before copulatory test. Training of males Males were trained individually with active females in oestrus in a transparent mating arena (60 x 30 x 18 cm) for 5 min. A male was considered sexually active when it attempted to mount any active female introduced into the cage. Only such males were used for subsequent experiments. The average mountings in a normal male were found to be about 8-10 in 5 min. The males, which mounted 3-5 times in 5 min, were considered as sexually sluggish. Treatment The male rats were divided into 7 groups of 6 each. Aqueous suspensions of PAM-F and PAM-R were freshly prepared every day and were given 1 g/kg orally in the volume of 1 ml/100 g. A. treatment: After recording 6 control readings over a 10 day period, rats were treated with PAM-F and PAM-R for 12 days. Test readings were taken 1 h after the drug treatment on the 4, 8 and 12 days. Group 1: Normal copulators PAM-F Group 2: Normal copulators PAM-R Group 3: Sluggish copulators PAM-F Group 4: Sluggish copulators PAM-R B. Chronic treatment: Drug schedule started in one-month-old rats for 66 days. When the rats reached 70 days of age, they were trained for sexual behaviour and tested on the days 92, 94 and 96. Group 5: Normal copulators Saline Group 6: Normal copulators PAM-F Group 7: Normal copulators PAM-R The following parameters were observed 13

3 (a) (b) (c) (d) (e) Mounting frequency: Number of mounts in a series or number of mounts in a given period of time (5 min.) Mounting latency: Time taken for the first mount from the introduction of the male into the mating arena. Ejaculation frequency: Number of ejaculations in a given period of time (5 min.) Ejaculation latency: Interval between the first mount to first ejaculation. Post-ejaculation interval: Time taken after ejaculation before resumption of sexual activity. Mounting latency was significantly decreased (p<0.05, 0.01, 0.001) in all the six groups compared to the control (Table 2). Among the test groups, mounting latency had significantly decreased (p<0.01) in both acute and chronic PAM-R treatment. Table 1: Effect of acute and chronic treatment of PAM-F and PAM-R on mounting frequency in male rats (n=6; Mean ± SE) 0 day 4 th day 8 th day 12 th day Normal PAM-F 8.10 ± ± ± ± 0.94 Normal PAM-R 8.58 ± ± ± 0.80** ± 0.76 Sluggish PAM-F 0.80 ± ± ± ± 0.83 Sluggish PAM-R 0.97 ± ± ± 1.07** ± 1.15** Control 0.74 ± ± ± ± 1.47 PAM-F 1.66 ± ± ± ± 1.45 PAM-R 4.35 ± 1.70** ± 0.87** ± 1.41** ± 1.01** and PAM-R were significant (p<0.05, 0.01, not shown in the table). PAM-R compared to PAM-F: **p<0.01 (acute: 8 th and 9 th day; chronic: 92 nd, 94 th and 96 th days). Table 2: Effect of acute and chronic treatment of PAM-F and PAM-R on mounting latency (in seconds) in male rats (n=6; Mean ± SE) 0 day 4 th day 8 th day 12 th day Normal PAM-F ± ± ± ± 5.00 Normal PAM-R ± ± ± 14.27** ± 7.16** Sluggish PAM-F ± ± ± ± Sluggish PAM-R ± ± ± ± 6.54** Control ± ± ± ± 9.46 PAM-F ± ± ± ± 8.73 PAM-R ± ± 15.10** ± 9.57* ± 3.97** and PAM-R were significant (p<0.05, 0.01, not shown in the table). PAM-R compared to PAM-F: **p<0.01 (acute: 8 th and 9 th day; chronic: 92 nd, 94 th and 96 th days). Ejaculatory frequency was significantly increased (p<0.05; 0.01) in all the six groups as compared to their respective controls (Table 3). There was no ejaculation in sluggish groups

4 before treatment. However, significant increase in ejaculatory frequency (p<0.01) was noticed after PAM-F and PAM-R treatment on 12 th day. Among treated groups either actual or chronic, difference in ejaculatory latency was not significant. Table 3: Effect of acute and chronic treatment of PAM-F and PAM-R on ejaculatory frequency in male rats (n=6; Mean ± SE) (Control) 4 th day 8 th day 12 th day Normal PAM-F 0.19 ± ± 0.21* 0.50 ± 0.22** 0.66 ± 0.21** Normal PAM-R 0.22 ± ± 0.21* 0.66 ± 0.21** 0.83 ± 0.16** Sluggish PAM-F ± 0.21** Sluggish PAM-R ± ± 0.21** Control ± ± 0.10 PAM-F ± 0.10* 0.33 ± 0.21* 0.50 ± 0.22* PAM-R ± 0.21* 0.50 ± 0.22* 0.83 ± 0.16* PAM-R and PAM-F compared to their predrug readings in acute experiments and control readings in chronic experiments: *p<0.05; **p<0.01 Significant increase (p<0.05; 0.001) in ejaculatory latency was seen in all the six groups as compared to the control (Table 4). Among acute test groups ejaculatory latency was significantly increased (p<0.01) in PAM-R groups. Among chronic test groups, significant difference in ejaculatory latency was observed in PAM-R group only on the last reading (96 th day). Table 4: Effect of acute and chronic treatment of PAM-F and PAM-R on ejaculatory latency (in seconds) in male rats (n=6; Mean ± SE) (Control) 4 th day 8 th day 12 th day Normal PAM-F ± ± ± ± 7.35 Normal PAM-R ± ± 8.00** ± 2.19** ± 2.54** Sluggish PAM-F ± Sluggish PAM-R ± 0.00** ± 4.56** Control ± ± 0.10 PAM-F ± ± 1.66 PAM-R ± ± ± 5.14* and PAM-R were significant (p<0.05, p<0.01 not shown) on testing dates. PAM-R compared to PAM-F: *p<0.05 (chronic: 96 th day); **p<0.01 (acute normal: 4 th, 8 th and 12 th days) and **p<0.01 (acute sluggish : 8 th and 12 th days) Significant decrease (p<0.05, p<0.001) in post ejaculation interval was seen in all the six groups compared to control (Table 5). Among the treated groups post ejaculation interval was significantly decreased (p<0.01) in both acute and chronic PAM-R groups. Table 6 enumerates the effect of PAM-F and PAM-R on body and sex organ weights (except liver) in male albino rats. Significant gain in body and organ weights (p<0.05; 0.01) were seen in both PAM-F and PAM-R treated groups compared to the control group. Body and sex

5 organ weights, after PAM-R treatment, were significantly (p<0.01) increased compared to PAM-F treatment. Table 5: Effect of acute and chronic treatment of PAM-F and PAM-R on post-ejaculatory interval (in seconds) in male rats (n=6; Mean ± SE) (Control) 4 th day 8 th day 12 th day Normal PAM-F ± ± ± ± Normal PAM-R ± ± 10.00** ± 4.76** ± 4.73** Sluggish PAM-F ± 5.00 Sluggish PAM-R ± 0.10** ± 5.46** Control PAM-F ± ± 4.33 PAM-R ± 7.50* ± 5.77** ± 5.00* and PAM-R were significant (p<0.05, p<0.01 not shown in the table). PAM-R compared to PAM-F: *p<0.05 (chronic: 92 nd day); **p<0.01 (acute normal: 4 th, 8 th and 12 th days) and (acute sluggish : 8 th and 12 th days) The effect of PAM-F and PAM-R on sperm count is enumerated in Table 6. Significant increase (p<0.01) in the number of sperms was seen in both PAM-F and PAM-R treated groups compared to the control group. PAM-R shows highly significant (p<0.01) effect on sperm count as compared to PAM-F. Table 6: Effect of chronic treatment of PAM-F and PAM-R on body, sex organs weights and sperm count in male rats (n=6; Mean ± SE) Groups Control PAM-F PAM-R Body weight ± ± ± 8.06 Testis 1.34 ± ± ± 0.08 Seminal vesicle ± ± 9.64 Ventral prostate ± Adrenal gland ± ± ± Cauda epididymis Liver 9.88 ± ± ± 0.06 Compared to control, PAM-F and PAM-R were significant (p<0.05; p<0.01 not shown in the table) PAM-R compared to PAM-F: **p<0.01. Sperm count million/ml ± ± ± 1.83 A significant improvement of these parameters on sexual activity shows its beneficial effects in male impotence and sterility. Increase in mounting frequency and mounting latency indicates that PAM-R along with increasing libido, probably increases the potency. The increase in ejaculatory latency shows that PAM-R prolongs the duration of coitus. The increase in ejaculatory latency as well as decrease in post-coital interval indicates that PAM-R intensifies sexual activity in a sustained manner. The earlier reports on the Unani preparations suggested that the improved sexual function could partially be due to androgenic activity 16. Mithra et al. (1996) found increased testosterone levels after PAM-F and PAM-R treated groups using RIA method. Histological examinations in their studies confirmed the higher cell count of the LH-FSH.

6 Weight of accessory organs In a separate set of animals, chronic treatment schedule was given as described in the groups 5-6 and the animals were weighed daily before treatment. At the end of the experimental period the animals were anaesthetized with pentobarbitone 40 mg/kg i.p. and testis, seminal vesicle, ventral prostate, cauda-epididymis and liver were dissected out, blotted and weighed on a torsion balance 14. Sperm count The cauda epidydymides was punctured and the fluid was collected in an RBC pipette and diluted to 10% with phosphate buffered saline (ph 7.1). The suspension was filtered through an 80-um stainless steel mesh to remove tissue fragments 15. A fraction of this suspension was diluted (1:9) with the diluting medium. The sperm count was done on a Neubauer s chamber 16. Statistical analysis Sexual behaviour activity on different days was analysed by paired t test to calculate the level of significance. Results of different groups on the same day was done by ANOVA. The level of confidence was fixed at 95%. Comparison of body weight, organ weight and sperm count were done by ANOVA. RESUTLS AND DISCUSSION Mounting frequency was significantly (p< ) improved after acute and chronic treatment of PAM-F and PAM-R in normal and sluggish copulators (Table 1). Among the treated groups, mounting frequency was significantly producing basophils with reduction in the population of ACTH producing cells in both the treated groups. In addition the improvement in sperm count indicates the increased activity of FSH producing cells. These herbal products probably act at LHRH and FSHR levels to bring about a rise in the hormonal levels. Further it was also proved that both herbal preparations reversed the inhibitory effect of alcohol on sexual behaviour. The hormonal effects are reversible once the drug is withdrawn 10. Thus, PAM-R can be used clinically for improving cases of depressed libido and potency, of both organic and psychogenic origin. It is also indicated for the management of premature ejaculation without fear of side effects. REFERENCES 1. Beach, F.A., Psychosom. Med., 1942, 4, Ahlenius, S. Engel, J., Eriksson, H. and Sodersten, P., J. Neuroal Transm., 1972, 33, Malmnas, C.O., Adv. Biochem. Psychopharmacol., 1974, 11, Meyerson, B.J., Acta Physiol. Scand., 1964a, 63, 241p. 5. Meyerson, B.J., Psychopharmacologia., 1964b, 6, Meyerson, B.J., Arch. Int. Pharmacodya., 1964c, 150, Zemlan, F.P., Ward, I.L., Crowley, W.R. and Margules, D.L., Science, 1973, 179,

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