Shilajit for Men Health
Share
Clinical evaluation of purified Shilajit on testosterone levels in healthy volunteers
Summary
Purified Shilajit, an Ayurvedic rasayana, was evaluated in healthy volunteers of age between 45 and 55 years for its effect on male androgenic hormone viz. testosterone in a randomised, double-blind, placebo-controlled clinical study at a dose of 250 mg twice a day. Treatment with Shilajit for consecutive 90 days revealed that it has significantly (P < 0.05) increased total testosterone, free testosterone and dehydroepiandrosterone (DHEAS) compared with placebo. Gonadotropic hormones (LH and FSH) levels were well maintained.
Introduction
Purified Shilajit (PS) is used in Ayurveda, indigenous system of Indian medicine, as a remedy for several diseases, particularly chronic diseases. Shilajit is a pale-brown to blackish-brown exudate that oozes from sedimentary rocks worldwide, largely in the Himalayas. Common people describe it from their knowledge as pahar-ki-pasina (sweat of mountains), paharki-khoon (mountain blood), shilaras (rock juice), asphalt, bitumen, etc. Shilajit is said to carry the healing power of these great mountains (David & Vasant, 2001). It is an important drug of the ancient Ayurvedic materia medica and it is to this day used extensively by Ayurvedic physicians for a variety of diseases. Early Ayurvedic writings from the Charaka Samhita (Sharma, 1998) describe Shilajit as a cure for all diseases as well as a Rasayana (rejuvenator) that promises to increase longevity. It is composed of rock humus, rock minerals and organic substances that have been compressed by layers of rock mixed with marine organisms and microbial metabolites (Ghosal, 1994).
Traditional uses of Shilajit primarily focus not only on diabetes and diseases of the urinary tract, but also on oedema, tumours, muscle wasting, epilepsy and even insanity. Modern indications extend to all systems of the human body with a significant number of additions in the reproductive and nervous system. Clinical research confirms many of the properties for which Shilajit has been used (Talbert, 2004). In Ayurveda, Shilajit is employed for the management of male reproductive disorders, and in particular, under the parlance of Vrisya (an aphrodisiac with special reference to spermatogenesis) (Sharma, 1998).
Several toxicological studies, both acute and subchronic, have already been performed by many scientists with Shilajit throughout the world. Oral LD50 was found to be >2000 mg kg−1 (Acharya et al., 1988; Ghosal et al., 1989), and Shilajit was proved to be safe at doses of 0.2–1.0 g per kg body weight when used chronically (Kelginbaev et al., 1973; Anisimov & Shakirzyanova, 1982; Fortan & Acharya, 1984; Al-Hamaidi & Umar, 2003). Clinical evaluation of spermatogenic activity of processed shilajit in oligospermia (Biswas et al., 2009) revealed that there was no alteration on objective features related to any systemic toxicities such as serum urea, uric acid, serum bilirubin, total protein, serum globulin, SGPT, SGOT and alkaline phosphatase (Biswas et al., 2009). Besides, it was also observed that there was a significant (P < 0.001) improvement in spermia (+37.6%), total sperm count (+61.4%), motility (12.4–17.4% after different time intervals), normal sperm count (+18.9%) and total testosterone (+23.5%) with concomitant decrease in pus and epithelial cell count compared with baseline value in 28 patients of oligospermia after 90 days of treatment with PS at a dose of 100 mg twice daily. An unpublished human safety study of purified Shilajit from Natreon, Inc., New Brunswick, NJ, USA by the present authors has demonstrated the safety of this product at 250 mg twice a day dosing (Jana, Utpalendu, Biswas, Tuhin; J.B.Roy Ayurvedic Medical College and Hospital, Kolkata, India, Project No.: JBR/Res/01/2007), and an unpublished animal study in rats with 100 mg/kg b.w. (equivalent to human dose of 850 mg) by Natreon showed a significant increase in testosterone levels. Thus, a dose of 250 mg twice daily was selected for this study.
Materials and methods
Preparation and analysis of the drug
Purified Shilajit (PrimaVie™, Kolkata, West Bengal, India), a patented (US 6,440,436, 6,969,612, 6,558,712, EP 1 387 614) standardised extract of native Shilajit from Natreon, Inc., was used for the study. Shilajit was standardised to contain not less than 60% w/w of total bioactives, which include not less than 50% w/w of fulvic acids (FAs), not less than 0.3% w/w of dibenzo α-pyrones (DBPs) and not less than 10% w/w of dibenzo-α-pyrone chromoproteins (DCPs), as quantified by HPLC using external standards (isolated from Shilajit extract by low pressure (Lobar) chromatography). An HPLC chromatogram of this product is shown in Fig. 1.
Selection of healthy volunteers
The clinical trial was conducted between December 2012 and March 2014 at J. B. Roy State Ayurvedic Medical College and Hospital, Kolkata, India, after obtaining necessary permission from Institutional Ethics Committee (IEC) of J. B. Roy State Ayurvedic Medical College and Hospital, Kolkata, India. A schematic of the study design is shown in Fig. 2. Healthy volunteers aged between 45 and 55 years, irrelevant of religion, income status and occupation, were selected for the present purpose, and the distribution of patients was done by the method of double-blind randomised techniques. Initially, 145 volunteers were selected on the basis of primary assessment eligibility and 49 among them were excluded for various reasons (Fig. 2). A total of 96 volunteers were enrolled in the present trial and randomly divided into two equal groups as PS treated and placebo treated, each with 48 subjects. In the course of the study, 21 subjects discontinued for various reasons and 38 subjects in PS-treated group and 37 subjects in the placebo group completed the study (Fig. 2). Mean age of the volunteers was 49.44 years in the test drug group and 48.89 years in the placebo group, and thus, there is no bias due to the difference in mean ages. Volunteers were included in the study after taking their consent in trilingual (English, Bengali and Hindi) using prescribed format of WHO-Helsinki rules. History of volunteers was taken according to the standard protocol mentioning name, age, sex, religion, address, occupation, income status, history of past illness, family history, personal history, marital history, general examination, systemic examination, laboratory investigation, etc.
Dosage regimen
- Group – I: PS 250 mg BID (38 subjects)
- Group – II: Placebo 250 mg (microcrystalline cellulose 124 mg + lactose 124 mg + magnesium stearate 2 mg) BID (37 subjects)
Both the PS and placebo capsules were white opaque, Size 1, and looked identical.
Inclusion/exclusion criteria
Eligibility was based on the following inclusion criteria: Clinically examined normal healthy volunteers, devoid of any chronic, organic or dreadful diseases, irrespective of religion and income status with age between 45 and 55 years, not taking any supplements or vitamins and male subjects. The exclusion criteria were volunteers of age below 45 years and above 55 years, any chronic, organic or dreadful diseases, concomitant serous disorders, other drug treatment being received simultaneously on long basis which may affect the study results, any immunosuppressive drugs being received, poor nutritional status, subjects likely to fail compliance with the trial protocol, alcoholism, smoking, using antipsychotic drugs and steroids/male contraceptives.
Observation criteria
Screening and diagnosis
Ageing men often present with a variety of vague nonspecific symptoms that may be associated with testosterone deficiency (Nandy et al., 2008). Subjects were, therefore, screened on the basis of the St Louis University ADAM questionnaire (Table 1) (Morales et al., 2000). Besides the ADAM questionnaire, normalcy of the subjects was established after receiving acceptable ranges of different haematological and biochemical parameters such as fasting glucose, serum urea, creatinine, ALT, AST, Hb%, total RBC, total and differential counts of WBC, RBC/WBC morphology, ESR and routine stool examination including occult blood test. Biochemical investigations were performed by means of auto-analyser (Beckman Coulter, CA, USA), and haematological investigations were done by means of automated cell counter (Medonic CA 530 16; Oden (Merck, Germany). Routine stool examination was carried out by means of conventional methods.
Estimation of testosterone, free testosterone, LH, FSH and DHEAs
Both total and free testosterones, LH, FSH and DHEAs were estimated from fasting blood of each volunteer on days 0 (baseline), 30, 60 and 90. LH and FSH were estimated by means of ADVIA Centaur XP Immunoassay Systems (SIEMENS, Berlin, USA) with ADVIA Centaur LH ready pack primary reagent; testosterone and DHEAs were estimated by means of Access Immunoassay System (Beckman Coulter) with Access Testosterone Reagent Pack and Access DHEAs Reagent Pack respectively. Free testosterone was estimated by means of AccuBind ELISA Microwells Test system (Monobind Inc., Lake Forest, CA, USA) with AccuBind free testosterone reagent pack.
Statistical analyses
Statistical analysis of data of the two treatment groups collected at different intervals of study was performed by paired Student's ‘t-’test using spss11.5 version software (Chicago, USA).
Results
Subject screening
All subjects responded ‘yes’ to questions 1 and 7 or any three other questions according to the ADAM questionnaire were primarily selected. Besides, they also fulfil all inclusion criteria. Fasting glucose, renal investigations like serum urea and creatinine, hepatic investigations like ALT and AST, haematological parameters and stool investigations were found within normal range during primary screening for the inclusion of the volunteers.
Treatment efficacy
A total of 21 subjects discontinued the study, and any data from these subjects were not included in the calculations as these subjects discontinued at different time points of the study and for various reasons.
Testosterone and free testosterone estimation
It was observed that in PS-treated group, there was an increase in testosterone levels (ng ml−1) on days 30 (6.82%), 60 (3.09%) and 90 (20.45%) with respect to day ‘0’. The increment of testosterone levels on day ‘90’ was significant (P < 0.05) with respect to the values of day ‘0’. In placebo-treated group, there was a significant (P < 0.05) decreasing trend of testosterone level. The level of testosterone in PS-treated group on day 90 was found to be significantly (P < 0.05) better than the values of placebo-treated group in same day (Table 2). The level of free testosterone (pg ml−1) in PS-treated group on day 90 (19.14%) was significantly better (P < 0.05) than 0 day value and maintain parity with the testosterone level. Free testosterone level of PS-treated group on day 90 was also found to be significantly higher (P < 0.05) with the values of placebo-treated group on same day (Table 2).
Discussion
Testosterone is an anabolic steroid synthesised primarily by the Leydig cells in the testes in males, the ovaries in females and adrenal glands in both sexes. It is synthesised from cholesterol, with androstenedione, androstenediol, dehydroepiandrosterone sulphate (DHEAs), progesterone and pregnenolone acting as some of the intermediate substrates. Testosterone production is regulated by hormonal secretions from the hypothalamus and the pituitary gland in the brain via hypothalamus–pituitary–testicular axis. The process begins as the hypothalamus secretes gonadotropin-releasing hormone (GnRH) in generative pulses. In response to these steady intermittent bursts of GnRH, the pituitary gland releases luteinising hormone (LH) and follicle-stimulating hormone (FSH), which act directly on the testes. FSH activates the Sertoli cells that produce sperm (spermatogenesis). LH stimulates the Leydig cells to secrete testosterone in a daily rhythm characterised by peak levels in the morning and low levels in the evening. Once it reaches high levels, testosterone production generates negative loop feedback to the hypothalamus to downregulate LH release and diminish further testosterone production. In this way, testosterone inhibits its own secretion (Gingrich, 2010). In addition to hypothalamic influence, it has been found that testosterone has direct negative feedback effects on the anterior pituitary gland. Like most hormones, testosterone is supplied to target tissues in the blood where much of it is transported bound to a specific plasma protein, sex hormone-binding globulin (SHBG) (Brooks, 1975). Most circulating testosterone is bound by SHBG and albumin; approximately 2% of total testosterone is free (not bound to protein). SHBG-bound testosterone is so tightly bound that it is not biologically active. Both free and albumin-bound testosterones are biologically active and together are referred to as the bioavailable fraction (Bhasin et al., 2010). It is reported that the total, free and bio-available testosterone varies widely in the age group between 18–69 and 70–89 (Rosner et al., 2007). In the current study, total and free testosterones were estimated to establish the efficacy of PS for its role on testosterone stimulation and secretion property considering the reference range 4–30 pg ml−1 for free testosterone and 1.75–7.81 ng ml−1 for testosterone (total testosterone). DHEAs, the main precursor of testosterone, was also estimated to rationalise the role of PS on testosterone synthesis considering reference range 70–495 μg dl−1 between the age group of 41–50 years and 38–313 μg dl−1 between the age group of 51–60 years. The present research work reveals that PS may be able to increase both total testosterone and free testosterone with respect to baseline value indicating its potentiality on testosterone secretion level. The significant betterment of DHEAs with the treatment of PS signifies its role on testosterone synthesis. Other two gonadotropic hormones, viz. LH and FSH, were studied in this present work, to rationalise the hypothalamo-pituitary–testicular axis, where both of these hormones were in maintained levels indicating their initial role of triggering of testosterone production. This was followed by downregulation of LH and FSH on one hand and maintenance of the hypothalamo-pituitary–testicular axis by means of elevated level of testosterone on 30, 60 and 90 days on the other hand. All these modus operandi of PS on synthesis and stimulation of testosterone are found to be better than placebo-treated group in healthy male volunteers in age group of 45–55 years, who may undergo andropause in normal course. Placebo, composed with microcrystalline cellulose, lactose and magnesium stearate, has neither stimulation nor inhibiting role on testosterone secretion or synthesis.
Processed Shilajit (PS) containing biologically active component di-benzo-alpha-pyrone (DBP) is earlier reported to increase the spermatogenic activity in selected patients of oligospermia (Biswas et al., 2009), and the current study demonstrates that purified Shilajit increases the total and free testosterone levels in healthy volunteers.
Conclusion
The present study was conducted to evaluate the efficacy of PS for testosterone secretion and stimulation effects on normal healthy volunteers in the age group of 45–55 years. This effect was clarified by estimation of free and total testosterone on 0, 30, 60 and 90 days where the rise of these two androgenic markers was significant. Testosterone synthesis and secretion was supported by the maintenance levels of two gonadotropic hormones LH and FSH as well as elevation of testosterone precursor DHEAs.
Acknowledgements
We acknowledge Natreon Inc, Premise No: 02-360, Rishi Tech Park (Ground Floor), Action Area 1D, Newtown, Kolkata 700 156, for financial support to carry out the study.
References
- Acharya SB, Fortan MH, Goel RK, Tripathi SK, Das PK (1988) Pharmacological actions of Shilajit. Indian J Exp Biol 26: 775–777.
- Al-Hamaidi AR, Umar M (2003) Safe use of Shilajit during pregnancy of female mice. Online J Biol Sci 3: 681–684.
- Anisimov VE, Shakirzyanova RM (1982) Application of Mumie in therapeutic practice. Kazan Med Zh 63: 65–68.
- Bhasin S, Cunningham GR, Hayes FJ, Matsumoto AM, Snyder PJ, Swerdloff RS, Montori VM (2010) Testosterone therapy in men with androgen deficiency syndromes: An Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab 95: 2536–2559.
- Biswas TK, Pandit S, Mondal S, Biswas SK, Jana U, Ghosh T, Tripathi PC, Debnath PK, Auddy RG, Auddy B (2009) Clinical evaluation of spermatogenic activity of processed shilajit in oligospermia. Andrologia 42: 48–56.
- Brooks RV (1975) Androgens. Clin Endocrinol Metab 4: 503–520.
- David F, Vasant L (2001) The Yoga of Herbs. Lotus Press, Twin Lakes, WI. 250.
- Fortan MH, Acharya SB (1984) Pharmacological studies of Shilajit. Indian J Pharmacol 16: 45.
- Ghosal S (1994) The aroma principles of Gomutra and Karpuragandha Shilajit. Indian J Indg Med 11: 11–14.
- Ghosal S, Lal J, Sing SK, Dasgupta G, Bhaduri J, Mukhopadhyay M, Bhattacharya SK (1989) Mast cell protecting effects of Shilajit and its constituents. Phytother Res 3: 249–252.
- Gingrich JR (2010) Pathophysiology Course – Endocrine Module, Male Gonadal Disorders, Testicular Disorders & Clinical Conferences, December 6, 2010.
- Kelginbaev NS, Sorokina VA, Stefandu AC, Ismailova VN (1973) Treatment of long tubular bone fractures with Mumie and preparation in experimental and clinical conditions. Exp Surg Anesth 18: 31–35.
- Morales A, Heaton JP, Carson CC (2000) Andropause: a misnomer for a true clinical entity. J Urol 163: 705–712.
- Nandy PR, Singh DV, Madhusoodanan P, Sandhu AS (2008) Male andropause: a myth or reality. MJAFI 64: 244–249.
- Rosner W, Auchus RJ, Azziz R, Sluss PM, Raff H (2007) Position statement: utility, limitations, and pitfalls in measuring testosterone: an Endocrine Society Position Statement. J Clin Endocrinol Metab 92: 405–413.
- Sharma PV (1998) Charaka Samhita. Chowkhambha Orientalia Chikitsasthana, Varanasi, Karaprchitiya RasayanaPada, 4th edn. vol 2, verse no. 49–50. Chowkhambha Orientalia Chikitsasthana, Varanasi, U.P., India.
- Talbert R (2004) Shilajit: A Materia Medica Monograph, A paper submitted in partial fulfillment of the requirements for the degree of Clinical Ayurvedic Specialist at California College of Ayurveda, 1117A East Main Street, Grass Valley, California.