A Pilot Study on Evaluation of Standardized Ayurveda formulation Ashwagandhadi lehya as Aphrodisiac and in treatment of Oligospermia

 

Manan Raval1, Manish Patel2, Kanan Gamit1, Kalapi Patel2, S. N. Gupta2

1Pharmacognosy and Phytochemistry Department, Ramanbhai Patel College of Pharmacy, Charusat,

At and Post – Changa, Dist. Anand, Gujarat, India. 388421

2J.S. Ayurveda College, College Road, Nadiad, Gujarat, India. 387001

*Corresponding Author E-mail: mananraval.ph@charusat.ac.in

 

ABSTRACT:

Ashwagandhadi lehya is traditionally used as aphrodisiac and spermatogenic. It is official in Ayurvedic Formulary of India. Root powder of Withania sominfera is incorporated as one of major ingredients in the formulation. The studies were aimed to evaluate Ashwagandhadi lehya, Clincially, as aphrodisiac and in treatment of Oligospermia. The duration of studies was 90 days and patients showed willingness to participate in studies were asked to submit blood and serum samples on 0, 45th and 90th day. Alteration in serum FSH, serum LH, serum testosterone as well as alteration in sperm concentration, % of normal sperms and % motile sperms in semen samples were selected as evaluation parameters to assess role of Ashwagandhadi lehya in treatment of oligospermia. Aphrodisiac potential of the formulation was evaluated by recording the responses of patients in a questioner, prepared on the basis of International Index of Erectile Function (IIEF- 5), included their experience in sexual desire, erection, penile rigidity, ejaculation and orgasm before and after treatment.  The results showed that mean serum concentration of testosterone was increased by 8.31 % after 45 days of treatment and 16.53 % after 90 days of treatment, without concurrent alteration in serum FSH and serum LH concentration. Semen analysis showed that, after 90 days of treatment, mean sperm concentration in semen was increased by 200%, while % of normal sperms and % motile sperms were found to be increased by 11% and 59 %, respectively. The results of the studies showed, statistically significant improvement in sexual arousal, penile rigidity, orgasm and ejaculation, after treatment. As, testosterone concentration was increased in absence of increased LH release, it was concluded that formulation might act directly on testis and stimulated the expression of the proteins responsible for testosterone synthesis. It was also proposed that, increased in serum testosterone level might be responsible for aphrodisiac potential of Ashwagndhadi Lehya

 

KEYWORDS: Ashwagandhadi lehya, Vajikaran, Oligospermia, Spermatogenic, Aphrodisiac, Withania somifera.

 

 


INTRODUCTION:

Infertility is believed to be one of the major reproductive health concerns. As per the WHO report, around 6080 million couples worldwide suffer from infertility and male factor contributed to 50% of these cases. Abnormalities in semen or in sperms were reported to be considered for male factor infertility1,2.

 

However, overall human male fertility has not declined, but there were evidences showed, deterioration in human semen parameters worldwide3,4,5.

 

Male sexual dysfunction (MSD) was suggested to be disorder associated with a wide range of physical and psychological conditions. The prevalence of sexual dysfunction was found in 31 % male respondents in a study. About 10 % of respondents confirmed trouble in achieving or maintaining erection, about 15 % were found to possess low sexual desire and approximately 8% had inability to achieve orgasm6,7,8,9.

 

Ashwgandhadi lehya (AL) is used traditionally as aphrodisiac and in treatment of impaired functions of male reproductive system10. It is official in Ayurvedic Formulary of India. AL is a semi-solid poly herbal preparation, prepared with the addition of sugar or sugar candy to the plant powders assimilated with butter oil by constant heating and stirring. It was reported to contain equal proportion, each of powdered roots of Withania somnifera (WS), roots of Hemidesmus indicus, fruits of Cuminum cyminum, roots of Smilax glabra, dried fruits  of Vitis vinifera. The standards to ensure quality of the AL are mentioned in Ayurvedic Formulary of India11. Amount of  withaferin -A(WA) was estimated from AL using HPTLC. Physical and chemical standards for AL were also evolved12. AL showed Rasayana effect in elderly patients, included beneficial effect on selected aging parameters in elderly subjects13,14. One of the major ingredients of AL is dried powdered roots of Withania sominfera (WS)15,16. Apart from other uses, WS is used as aphrodisiac, traditionally, and in treatment of seminal debilities17,18. Hydroalcoholic extract of WL root powder improved semen parameters, when evaluated in albino rats19. The aqueous extract of the roots of WS showed marked impairment in libido, sexual performance, sexual vigour and penile erectile dysfunctionin rats20. WS, when administered in form of extract protected swimming-induced reproductive endocrine dysfunctions in male rats21. Aqueous extract prepared from the roots of WS, stimulated spermatogenesis in rats. The spermatogenic potential was thought to be attributed to increased interstitial cells stimulating hormone and testosterone-like effects22. Controlled clinical studies in oligospermic patients showed that, WS root extract significantly improved deteriorated semen parameters and serum testosterone level as compared to the placebo treated patients23. WL inhibited lipid peroxidation and improved sperm count in infertile men. It also recovered the seminal levels of antioxidant enzymes24.

 

Current main line therapy of western medicine offers few remedies for treatment of oligospermia as well as to enhance the suppressed libido in male, but not without major side effects, and with arguable efficacy25-30. Many medicinal plants; mineral drugs and formulations, thereof, including AL are used traditionally in India, as aphrodisiac and in treatment of seminal debilities. Many of them have been subjected to pre-clinical evaluation, but few such plants or formulations were subjected to clinical evaluation on male reproductive functions. Though, AL was subjected to physical and chemical standardization, it has never been subjected to clinical evaluation as aphrodisiac and in treatment of oligospermia. The research activities, thus, were planned to evaluate the potential of AL as aphrodisiac as well as in treatment of oligospermia, in limited number of patients.

 

MATERIAL AND METHODS:

Procurement of AL:

AL was prepared (as 10 kg batch on our request) under expert supervision in Sunder Ayurveda teaching pharmacy of J. S. Ayurveda College, Nadiad. This manufacturing facility functioned as per the norms of GMP for manufacturing of Ayurveda products.

 

Procurement of Chemicals:

All the chemicals used were of AR Grade and procured from  Loba Chemie LTD, India. WA was received as gift sample from  Pharmanza Herbals PVT LTD, Dharmaj, Gujarat, India.

 

Standardization of AL:

AL was subjected to standardization process, by adopting procedure given in compendia11 (Ayurvedic Formulary of India, 2000).

 

Development of Chromatographic Fingerprint of AL and Estimation of WA:

Preparation of Sample Solution:

25 g AL was loaded on 100 g silica (Silica for Column Chromatography, 60-120). The mixture was extracted for 2 hr at 60°C using 200 ml hexane in a soxhlet extractor. Hexane extract was discarded. Dried marc was extracted using soxhlet extractor, with 200 ml methanol at 65°C for 4 hr. Methanol extract was collected, filtered   and concentrated using rotary vacuum evaporator at 40°C. Methanol was evaporated from concentrated mass on water bath. Semisolid, black- brown extract was preserved in desiccator at room temperature.

 

Preparation of Standard Solution:

Accurately weighted, 100 mg WA was dissolved in 50 ml methanol using laboratory sonicator. The solution was diluted up to 100 ml using methanol to produce standard stock solution. Six different aliquots of standard stock solution were diluted using methanol to produce different dilutions of WA, covered the range of 1 µg/ml to 200 µg/ml WA.

 

Construction of Calibration Curve and estimation of WA from sample:

Calibration range selected for the set of experiments was 1µg/ml to 200 µg/ml. Each concentration was injected for six times (n=6). Calibration curve was constructed by plotting average peak area versus corresponding concentration. The line equation was constructed using least square regression analysis. Amount of WA present in AL was calculated from line equation using area of peak corresponded to WA.

 

Optimized Chromatographic Conditions:

Chromatographic separation was achieved using Waters quadra gradient chromatographic system equipped with PDA detector. Optimized mobile phase was acetonitrile: water contained 4% v/v Glacial Acetic Acid (50:50 v/v). Column used was Phenomenex (250 X 4.6 mm, 5 µm) with flow rate set at 2 ml/min. Injection volume was 20 µl and chromatogram was recorded at 425 nm.  Data was collected, processed and integrated using Waters Empower software. The chromatogram of methanolic extract of AL with peak corresponded to WA is shown in Fig.1(a). The peak of WA from standard solution is shown in Fig.1(b).

 

Evaluation of AL:

Approval of Studies and Enrollment of Patients:

The research followed guidelines of the Declaration of Helsinki and Tokyo for humans, and the study protocol was approved by the human ethics committee of J. S. Ayurveda College – Nadiad. The approval number was JSAM/IECHR/19/03-2014. The subjects were introduced to the studies and counseled verbally before their enrolment. Written Informed Consent was collected from each subject before enrolling him in the studies.

 

Following were the set Inclusion–Exclusion criterions for the studies:

Inclusion Criteria:

1.    Infertile Male Patients aged between 21 - 45 Years

2.    Sperm concentration was less than 15 million Sperm/ ml31and not receiving any other treatment from outside and willingness to give written informed consent for participation in the study. Patients with complain of suppressed libido along with the above mentioned semen abnormality were preferred.

 

Exclusion Criteria:

1.    Any Concomitant disorder of vital organs.

2.    Received any infertility treatment for past one month and/or any other treatment expected to receive during the study period.

3.    Patients suffering from azoospermia, asthenospermia, necrozoospermia and teratozooaspermia

4.    Patients with history of mums, measles, small pox, chicken pox, tuberculosis or injury to genitalia in last 10 years.

5.    Patients diagnosed with diabetes, abnormal thyroid functions, malignancy, arthritis, sexually transmitted disease and/or consuming medication daily for any reason, were excluded from the studies.

6.    The subjects underwent any other surgical operative procedures which proved to be affecting reproductive function as well as diagnosed with varicocele were also excluded from the studies. 

7.    The patients with habit of consuming tobacco in any form, smoking or consuming alcohol were excluded.

 

The subjects opted to be enrolled for the studies were examined, before enrolment, for exclusion criterions. A total of fifty subjects with preliminary infertility and total sperm count less than 15 million/ ml and/or complain of suppressed libido showed willingness in Written Informed Consent with signature were included in the studies

 

 

 

Study Protocol:

Subjects who fulfilled the inclusion criteria were selected from the out-patient and in-patient department of P. D. Patel Ayurved Hospital attached with J. S. Ayurvea College, Nadiad, after receiving their consent. All enrolled patients were assigned to a group. Being single arm study, the group would serve as its own control; the comparison was made between before and after treatment period on single subject group. All the patients were instructed to take 10 g formulation twice daily before 1 h of principal meals, which would be continued for 90 days. All the patients assessed clinically before (initial level, 0 day), during the treatment (on 45th day) and on the day of completion of treatment (on 90th day).

 

Semen sample was collected from subjects after 3 days of sexual abstinence. The semen sample was collected in laboratory by masturbation and analyzed as per WHO guideline for semen analysis within 1 h time period. Venous blood sample was withdrawn from each subject on the same day. Following examinations were performed on 0-45th -90th day of treatment--

1.    Complete Blood Count (CBC), serum GPT, serum GOT, serum bilirubin and serum creatinine

2.    Serum testosterone, serum LH and serum FSH was estimated using Fully Automated Random Access Immune Assay Analyzer (model no.  AIA 360) manufactured by TOSOH – Japan. The instrument functioned with the Unit Dose Cup Test System (kit) provided by the manufacturer.

3.    Semen analysis32.

4.    Questionnaire based on international index for erectile function (IIEF)33, which was modified later 34.

 

The modified questionnaire was used to evaluate effect of therapy on sexual performance. Alteration in the mean score value, on 45th day and on 90th day as compared to those with on 0 day was considered to access effect of therapy as aphrodisiac.

 

Statistical Analysis:

Statistical analysis was performed using MedCalc15.4 and MS excel. Variables were summarized as mean with standard deviation. The normality of distributions was checked with Kolmogorov-Smirnov test. All continuous variable data sets received on 45th day and on 90th day were compared with those obtained on the day of the treatment commencement, individually, using the student’s paired t-test to ascertain the statistical significance. p ≤ 0.05 (rejecting H0 – Null Hypothesis) was considered as statistically significant alteration. Categorical (Ordinal) variables in case of evaluation of aphrodisiac activity (scores of selected attributes) were assessed using Wilcoxon Test (Paired Test).

 

RESULTS AND DISCUSSION:

The formulation was standardized by adopting the procedures given in compendia11. Acid insoluble ash value for AL and extractive values for AL was found within the set limits. Other physical parameters e.g. water soluble ash value, alcohol soluble extractive value, water soluble extractive value, were also found within the limits. The results confirmed that, the raw materials used to manufacture the formulation were processed appropriately and amount of earthy matter was controlled. The microbial content was also tested and found within the limit set for respective micro organism. The tests conducted to estimate amount of heavy metals (Lead, Arsenic, Mercury, Tin) using atomic absorption spectroscopy, revealed that the concentration was lesser than sensitivity of instrument, so instrument could not be able to detect the presence of metals tested.

 

The chromatographic conditions were optimized to get sharp and Gaussian peak of WA as well as to resolve peak of WA from nearby peaks in HPLC. The chromatogram, as shown in Fig 1(a) shows peak of WA.

 

 

Fig 1 (a). Peak of WA in Methanolic  Extract of AL

 

 

Fig 1 (b). Peak of WA in working standard solution

 


Table. 1:  Serum Concentration of Selected Hormones at Set Time Points of Treatment

Mean Serum Testosterone (ng/dL) ± SD

Mean Serum FSH (mIU/ml) ± SD

Mean Serum LH (mIU/ml) ± SD

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

568.09 ± 170.68

615.32 ± 173.52*

662.00 ± 194.12 **

9.00 ± 3.84

9.50 ± 3.75

10.20 ± 3.84

6.11 ± 1.82

5.90 ± 1.92

6.05 ± 1.75

n=39. *p ≤0.05, **p≤0.01, ***p≤0.001

 

Table. 2:  Determination of Selected Semen Parameters at Set Time Points of Treatment

Mean Sperm Count (million/ml)

(Mean± SD)

Motility ( % Motile) (Mean ± SD)

Morphology (% Normal)

(Mean± SD)

Negative Fructose Content 

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

 0 Day

45th Day

90thDay

14.0 ± 6.27

37.20 ± 30.70**

42.05±

20.57***

38.22

± 23.25

54.17± 25.52**

61.09 ± 22.52*

80.09 ± 21.35

85.92 ± 11.25

89.19 ± 10.98**

09

06

03

n=39. *p ≤0.05, **p≤0.01, ***p≤0.001

 


This chromatogram might serve as chromatographic fingerprint for AL and possibly be useful as one of identifying parameters for AL. The amount of WA was determined using line equation after placing the value of area of WA peak obtained in chromatogram after injecting AL sample solution. The mean amount of WA was found to be 0.028 ± 0.001 mg % w/w in AL. Moisture  content was found  10.46 ± 0.10 % in AL. AL was jiggery based poly herbal matrix. The amount of WS was 16.5 % on weight basis in formulation, when calculated from recipe. This might be the reason for low concentration of WA. As, WA has never been evaluated for spermatogenic or aphrodisiac potential, it was considered as chemical marker for AL.

 

There were fifty patients enrolled for the studies. Thirty nine patients continued studies, these thirty nine patients were considered, while assessing the results. Out of thirty nine enrolled patients, fifteen patients were self employed while remaining patients had job. Eleven patients were graduated in respective fields and nine patients passed 10th standard examination. 4 patients were illiterate.

 

The studies involved, determining hematological parameters, which included, % Hb, RBC, WBC and differential counts along with estimation of serum creatinine, serum GPT analysis and serum random blood sugar. The studies were performed to monitor probable effects of the formulation on hematological, liver and kidney functions. The studies revealed no notable alteration in selected parameters, except minor alteration in serum creatinine and serum blood sugar level, ruled out possibility of formulation affecting hematological, kidney and liver functions.

 

Hormonal and semen analysis was performed to evaluate probable effect of AL on spermatogenesis. The result of hormonal analysis and semen analysis performed on 0, 45th and 90th day was shown in Table 1 and Table 2, respectively.

 

It was revealed from the results that, serum testosterone concentration was increased as treatment progressed; as compared to it was on the day of commencement of treatment. The alteration was found statistically significant in case of serum testosterone concentration, while it was not in case of serum FSH and serum LH concentration. The results of semen analysis showed that, as the treatment progressed, the mean of sperm concentration, % motile sperm and % of normal sperms in semen samples were found increased. The alteration was found statistically significant. There were nine patients, having absence of fructose in semen. At the end of treatment, out of those, 6 patients recovered and semen analysis showed positive fructose content. Aphrodisiac potential was evaluated by considering the rating assigned by the patients to the questions in questioner. The results, as shown in Table 3, revealed that, except orgasm related questions (except on 45th day), the mean value of score was found increased as compared to that on commencement of treatment, in statistically significant manner. 

 

As low concentration of FSH, LH and testosterone resulted in impaired spermatogenesis as well as oligospermia, concentration of these hormones were selected as parameters to evaluate action of AL35.  The results of the studies, represented in graphical form as shown in Fig 2, showed that mean serum concentration of testosterone was increased by 8.31 % after 45 days of treatment and 16.53 % after 90 days of treatment, compared to mean concentration of serum testosterone, on the day of commencement of the treatment. The alteration in testosterone concentration was statistically significant. The alteration was not statistically significant in case of serum LH and serum FSH level. It was reported in another independent study that, WS root powder administered in a dosage of 5 g/day for 90 days to normozoospermic infertile man resulted in significant increase in serum testosterone level. The results of our studies were in accordance with the results obtained for WS alone23,24.

 

LH is secreted from anterior pituitary under the influence of GnRH. It binds to membrane receptors expressed on Leydig cells and promotes synthesis of proteins in Leydig cells36which ultimately results in an increased conversion of cholesterol to pregnenolone by Leydig cells. Pregnenolone is the intermediate for biosynthesis of testosterone in Leydig cells37, The studies showed that, concentration of serum LH was not increased significantly, but testosterone concentration was increased. This might be attributed to activation of negative feedback mechanism due to increased concentration of serum testosterone. As, testosterone concentration was increased in absence of increased LH release, it was proposed that formulation might act directly on the Leydig cells and stimulated Leydig cells to express the proteins responsible for testosterone synthesis. Though, not included in evaluation parameters, but the possibility of stimulation to expression of Androgen Binding Protein (ABP) and LH receptor on leydig cell should also not to be overruled.

 

When mean value for key semen parameters, determined on 45th day and 90th day of treatment, compared to that on the day of commencement of treatment, alteration in parameters found statistically significant. Alteration in value of selected semen parameters after the treatment is shown in Fig. 2. Semen analysis showed that, after 90 days of treatment, mean sperm concentration in semen was increased by nearly 200%. Mean value of percentage of motile sperm was increased by 59.16% at the end of treatment. The results also showed that percentage of normal spermatozoa in samples was increased by 11.34% during this period.

 

Sertoli cells facilitate the progression of germ cell to spermatozoa through direct contact and by offering controlled environment necessary for their growth38. The series of experiments suggested that, during the process of spermatogenesis, conversion of step 1 to 7 having round spermatids was maintained, even the concentration of testosterone was lower than normal, provided normal concentration of FSH was maintained. Conversion of step 7 to 8, of spermatogenesis, proceeded at reduced efficiency, if testosterone concentration in reduced. Apart from that, it was also observed that, testosterone affected formation of a junction area, termed as ectoplasmic specialization (ES).

 


Fig 2. % Alteration in Selected Parameters 

Table. 3:  Assessment of Aphrodisiac Potential

Sexual Desire

(Mean ± SD)

Erection (Mean ± SD)

Penile Rigidity

(Mean ± SD)

Ejaculation  (Mean ± SD)

Orgasm (Mean ± SD)

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

0 Day

45th Day

90th Day

3.10 ±

 0.78

3.40 ±

0.82 **

4.12 ±

 0.75 ***

3.02 ±

 0.88

3.31 ±

 0.86 **

4.00 ±

0.79***

1.00 ±

0.86

1.78  ±

 0.78**

2.25 ± 1.18***

3.25 ±0.70

3.53 ±

 0.79*

4.00 ±

0.83***

4.11 ±

 0.95

3.07 ±

 0.97*

4.58 ±

0.62**

n=39. *p ≤0.05, **p≤0.01, ***p≤0.001

 


It was proved that, at low concentration of testosterone, process of spermatogenesis was proceeding at a reduced rate, primarily due to compromised attachment of spermatids with Sertoli cell processes, due to improper formation of ES and compromised attachment of spermatids at junctional area39. The present studies did not show statistically significant increase in serum FSH concentration, overruled the possible role of FSH in getting higher concentration of sperm in semen after treatment. Concurrent increase in serum testosterone level as well as sperm concentration in semen after the treatment, thus, suggested that AL might stimulate sperm number and percentage of normal sperms in semen samples, due to its ability to increase mean serum concentration of testosterone in patients during treatment period. A multivariate analysis suggested that, total motile spermatozoa to be the most clinically relevant semen parameter and considered  to be the best predictor of fertility in the general male population40. Seminal peptides were found responsible, at least partly, for motility of sperm. Seminal plasma neutral endo peptidase (NEP/CD10) and amino peptidase N (APN/CD13) were shown to adversely affect sperm motility. Bradykinin (BK), and other components of the kallikrein-kinin system found increased sperm motility when used at sub nanomolar concentrations. It was also demonstrated that reduced concentration of one endogenous NEP/APN inhibitor peptide called opiorphin resulted in reduced sperm motility41. It was, thus, proposed here that, AL might act on expression of such peptide/s synthesis too, and thereby, stimulated the number of motile sperms.

 

Aphrodisiac potential of formulation was evaluated by assessing the response received from the patient in a questioner on the day of commencement, on 45th and 90th day of treatment. The patients graded their own sexual desire, erection, penile rigidity, ejaculation and orgasm, by considering the conditions given in the questioner, at each selected time point. A number was associated with each choice, thus, the response was measured from 1 (the lowest) to 5 (the highest). The data of score obtained for each category on 45th day and 90th day was compared with that of obtained on 0 day. The results of the studies are shown in Table 3 and in form of % alteration, are shown in Fig. 2.

 

The results of the studies showed, marked improvement in sexual arousal, penile rigidity, orgasm and ejaculation, after treatment. It is believed that, NO functions as a neurotransmitter and NOS is present in the regions of the brain that regulate sexual functions. Testosterone increases nitric oxide synthase in the medial preoptic area (MPOA).  Administration of testosterone to castrated male rats increased the number of NO synthase- labeled neurons in MPOA. NO was found  capable of stimulating dopamine release in the MPOA, which in turn stimulated penile erection42,43. This mechanism might constitute a basis for explaining the role of testosterone in sexual arousal. Previous studies performed with testosterone supplement and withdrawal in eugonad men suggested that, men who have gone through normal puberty and who have not been affected by aging, testosterone played an important role in their sexual interest and associated sexual arousability. The controlled studies with testosterone replacement on male subjects with hypogonadism showed that sexual interest was reduced during testosterone withdrawal, consistent with testosterone being necessary for normal levels of arousal in male35.The studies on effects of testosterone withdrawal and replacement on genital response (erection) to sexual stimuli revealed that, testosterone replacement caused significantly more rigid and longer duration of erectile responses44. These observations suggested that, the improvement in the serum testosterone level might be responsible for the aphrodisiac potential of AL, as evaluated by measuring sexual arousal and penile rigidity. Human male arousal mechanism though has not been understood fully, but it was showed that central dopaminergic mechanism has important role in mediating sexual behavior and erection. Dopamine in the nigro striatal tract influences motor activity. It activates numerous motivated behaviors, including copulation in mesolimbic state. Dopamine secretion in the MPOA controls genital reflexes and sexual motivation. Serotonin is another neurotransmitter important in male arousal. Serotonin has inhibitory role in erection, except 5-HT2C activity. Serotonin binding with 5-HT1A facilitates ejaculation. Oxytocin too believed to possess facilitator role on sperm and egg transport by increasing smooth muscle contractility in the reproductive tracts45. The improvement in ejaculation component of questioner, might be due to ability of the formulation to interact with oxytocin synthesis and/or release. The improvement in orgasm score after treatment might be attributed to interaction of contents of AL with serotoninergic system. As, the studies did not involve evaluation of the components interacted directly or indirectly with oxytocic or serotoninergic system, these two possibilities could not be tested.

 

CONCLUSION:

It was concluded from the studies that, AL could increase sperm concentration and percentage of normal sperms in semen sample of oligospermic patients, due to its ability to improve serum testosterone concentration. It was also suggested that, AL possibly interacted with testosterone biosynthesis mechanism and thus, could increase serum concentration of testosterone. It was also proposed that, AL might affect the testicular synthesis of peptides, which had effects on sperm motility. Improvement in score for sexual desire, penile rigidity and penile erection, after treatment, were attributed to rise in testosterone concentration. The improvement in ejaculation component of questioner and improvement in orgasm score after treatment could not be explained with the set of the studies performed. The set of studies were performed as pilot studies. It was further suggested to expand the   studies to investigate the effect of WS, alongside AL as aphrodisiac and spermatogenic. Incorporation of new parameters e.g. determination of serum dopamine concentration, serum cortisol concentration and serum serotonin along with detailed analysis of type of sperm motility affected was also proposed. Preclinical experiments were also performed and they shall be communicated for publication as separate studies. As, WS was present as one of the ingredients of AL, we did not rule out the possible contribution of other medicinal plants present in the formulation towards aphrodisiac and spermatogenic activity of AL.

 

CONFLICT OF INTEREST:

The authors declare no conflicts of interest. The work carried out as academic research. 

 

ACKNOWLEDGEMENT:

The authors are thankful to the Principal, J. S. Ayurveda College for kind consent to carry out these collaborative studies. The authors are also thankful to Dr. Lal Hingorani, CMD, Pharmanza Herbals Pvt Ltd for providing gift sample of WA for analytical study purposes. Our thanks, due to Ms. Ishita Champaneri, Ms. Bhumi Dave and Ms. Krina Desai, all were PG students at Ramanbhai Patel College of Pharmacy, for their critical help in compiling and analyzing the datasets.

 

FUNDING:

This work was financially supported by Charotar University of Science and Technology (CHARUSAT), Changa, India. (Grant No. CHA/ADM/EA/14/02/218- dated 08/02/2014)

 

 

 

GLOSSARY/ABBREVIATION:

Sr.

No

Word Used In Manuscript

Description

1

AL

Ashwagandhadi Lehya: Multi component formulation which was subjected to clinical evaluation

2

Aphrodisiac

(Roots in Greek Language)

A food, drink, or other thing that stimulates sexual desire.

3

HPLC

High Performance Liquid Chromatography: This  is a system employed to separate and estimate WA from AL

4

Rasayana

(Sanskrit Word)

Rasāyana is defined as a therapeutic measure which promotes the longevity, prevents aging, provides positive health and mental faculties, increases memory, and impart resistance and immunity against diseases.

5

Vajikarana

(Sanskrit Word)

Vājīkaraṇa is a special branch of Āyurveda which is practiced to promote the health and sexual life of the healthy men as well as of the sexually weak patients.

6

WA

Withaferin-A: It is a steroidal lactone isolated from WS and other plants

7

WS

Withaniasomnifera: Botanical name of the plant, which was present in AL. It is also known as Indian winter cherry

 

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Received on 08.01.2019          Modified on 10.02.2019

Accepted on 01.03.2019        © RJPT All right reserved

Research J. Pharm. and Tech. 2019; 12(5):2383-2390.

DOI: 10.5958/0974-360X.2019.00399.8