Withania somnifera as immunomodulators:

Detail discussion on their traditional uses, mode

of action, and responsible chemical agent in
plant, doses, and toxicity.
PHG-830

SUBMITTED TO:
SIR SALMAN AHMED
SUBMITED BY:
NORIN MEMON

DEPARTMENT OF PHARMACOGNOSY
Table of contents
1 Introduction _________________________________________________________________ 2
2 Traditional uses of Withania somnifera ____________________________________________ 2
3 Withania somnifera as immunomodulator _________________________________________ 3
4 Mode of action _______________________________________________________________ 4
5 Phytochemistry _______________________________________________________________ 4
5.1 Chemical constituents ______________________________________________________ 4
5.1.1 Alkaloids: ____________________________________________________________ 4
5.1.2 Steroidal lactones: ____________________________________________________ 5
5.1.3 Steroids: ____________________________________________________________ 5
5.1.4 Salts: _______________________________________________________________ 5
5.1.5 Flavonoids: __________________________________________________________ 5
5.2 Chemicals responsible for immunomodulatory effects ____________________________ 5
6 Doses _______________________________________________________________________ 6
7 Toxicity _____________________________________________________________________ 6
8 References __________________________________________________________________ 7

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Withania somnifera as immunomodulators:
Detail discussion on their traditional uses,
mode of action, and responsible chemical
agent in plant, doses, and toxicity.
1 Introduction
Withania somnifera generally recognized as Ashwagandha or Indian Winter Cherry, a member
of family Solanaceae, is an important Ayurvedic medicinal plant because of its chemical
composition, Pharmacological and therapeutic uniqueness. It is indigenous to Mediterranean
countries of Africa, Southern Europe, Eastern Asian countries like Pakistan and India. In
traditional Indian Ayurvedic system of medicines W. somnifera is known as herbal Rasayana
(lengthening lifespan). It is used for the treatment of various ailments like fever, asthma,
arthritis, rheumatism, inflammation, tuberculosis, mental diseases, and other medical ailments.
Now a days the withania somnifera is popular because of its immunomodulatory effects i-e it
possesses such chemical constituents that cause modification of immune system like either they
cause stimulation of antibody formation or inhibition of WBC activity and this effects is known
as immunomodulation and the chemical substances that cause immunomodulation are called
Immunomodulators.

2 Traditional uses of Withania somnifera

In Asia W. somnifera roots are used for various diseases since 3000 years and the root extract
is used for the preparation of about 200 formulations in Indian folk medicine which are used
for various purposes like an aphrodisiac and for the cure of various ailments like Asthma,
inflammation, insomnia, anxiety, psoriasis, constipation, fatigue, weakness, impotence,
premature ageing, ulcers, and mental stress.

Traditionally plant roots are used to stimulate youthful vigour and strength, improve the vital
fluid production and blood cells which will further contribute in strengthening immune system.
The root extract also enhance cerebral health, improve brain ageing and locomotive syndrome.
It is also a good remedy for weakness, epilepsy, memory loss, and some neurodegenerative
disorders like spinal cord injury, Alzheimer, and Parkinson’s disease. Roots can be used as
Rejuvenating drug, tonic, Alternative pungent, astringent, Aphrodisiac, Phthisis according to
the Ayurveda system of medicine. According to the Siddha medicinal system it is effective in
fever, and inflammation while the Unaani system of medicine suggests that it is efficient in

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treating Asthma, bronchitis, leucoderma, Arthritis, emenagogue and Folklare system of
medicine suggest the roots effectiveness in cold, asthma, Tuberculosis and fever.

According to Ayurveda leaves are effective against Ulcers, painful swelling and tumors. In
Siddha system of medicine, leaves are effective against Fever, chest pain, sores, swelling.
Unani system suggest that leaves can be effective in External pains and as anti-inflammatory.
According to the folk medicine system, they are effective in treating cough, stress, nervous
exhaustion, memory loss, muscular weakness, syphilis, hemorrhoids, boils, eyesores and they
can be excellent diuretic and narcotic.

The seeds possesses anthelminthic, diuretic, narcotic and hypnotic properties and have ability
to removes corneal white spots. Since old times the fruits have been used as a topical cure for
tumors and tubercular glands, carbuncles, and skin ulcers.

3 Withania somnifera as immunomodulator

Besides all above therapeutic values of W. somnifera has a major effect on immune system as
it works as a general tonic to enhance the energy and protects body from diseases. In 1994
Grandhi et al performed a study that suggests, W. somnifera can be used as an immunological
adjuvant in the treatment of AIDs, cancer, and infection. Ziauddin et al performed an
observational study in 1996 upon cyclophosphamide, azathioprine and prednisolone treated
mice, in this study these animals were treated with W. somnifera extract and it was observed
that W. somnifera prevented myelosuppression which was primarily induced by above
immunosuppressive drugs, besides this a significant increase in the body weight, red blood cell
count, platelet count and HB value were also observed and these all observations give us a clear
clue about the immunomodulatory effects of W. somnifera extract. Another study suggests that
when cyclophosphamide treated animals are treated with W. somnifera extract then the chances
of cyclophosphamide induced leucopenia significantly reduced. One study suggests that
treatment of a mice with 20mg/kg of root extract of W. somnifera stimulated the immunological
activity. In year 2000 Davis and Kuttan analyzed mice treated with five doses of W. somnifera
extract and found that there was an increment in the bone marrow cellularity, WBCs count,
alpha, alpha esterase positive cell number, circulating antibody titer and number of plaque
forming cells in spleen and the extract had also inhibited the delayed type hypersensitivity
reaction and increased phagocytic activity of peritoneal macrophages. Singh et al suggest that
a polyherbal formulation named Immun-21 contains W. somnifera as its major constituent
possess significant immunomodulatory effects which may utilized for clinical benefits in case
of HIV Patients. Various research studies had suggested the immune-stimulatory potential of

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W. somnifera because of its actions on phagocytic activity of macrophages, hemolytic titers
and delayed type of hypersensitivity reactions. A study revealed that W. somnifera showed
immunomodulatory effects on cytotoxic lymphocytes production which resulted in sustained
tumor growth. In 2016 Rizvi et al performed a study where immunomodulatory effects of W.
somnifera were tested against Doxorubicin induced immunosuppression and it was observed
that W. somnifera caused an increase in WBCs count, absolute lymphocyte count and platelet
count after the suppression of bone marrow by Doxorubicin and it was also observed that bone
marrow cellularity and alpha esterase positive cell number were also increased. Rizvi et al
performed another study where they observed that the use of W. somnifera extract along with
antigen can cause an increase in the circulating antibody titer and plaque forming cells in
spleen.

4 Mode of action
Some studies suggest W. somnifera shows its immunomodulatory effects primarily by
increasing the levels of interferon (IFN)-γ, interleukin (IL)-2 and granulocyte macrophage
colony stimulating factor.

Another possible mode of action of Withania somnifera could be an increase in the expression
of Helper t cells 1, CD4, and CD8 counts and a dose dependent increment in natural killer (NK)
cell activity along with fastest recovery of CD4+ T cells in immune suppressed animals by
withanolides-A present in the roots of W. somnifera.

Some authors suggest that the stimulation of cell mediated immunity, IgM and IgG and
augmentation in proliferation and differentiation of lymphocytes according to surface markers
of T cells (CD3+, CD4+ and CD8+) and B cells (CD19+) could also be the main mode of
action of W. somnifera in immunomodulatory effects.

5 Phytochemistry
5.1 Chemical constituents
Phytochemical analysis of W. somnifera suggest that it possess different chemical constitutes
in different parts of plant and of which some major constitutes are discussed below. Studies
also suggest that a major portion of chemical constituents is based on more than 12 alkaloids,
40 withanolides and several sitoindosides.

5.1.1 Alkaloids:
Withanine, Withananine, Withasomnine, Somniferine, Tropeltigloate, Somniferinine,
Somninine and Nicotine.

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5.1.2 Steroidal lactones:

Withaferin-A, Withanone, Withanolide-E, Withanolide-F, Withanolide-A, Withanolide-G,

Withanolide-H, Withanolide-I, Withanolide-J, Withanolide-K, Withanolide-L, Withanolide-
M.

5.1.3 Steroids:
Cholesterol, B-Sitosterol, Stigmasterol, Diosgenin, Stigmastadien, Sitoinosides VII,
Sitoinosides VIII, Sitoinosides IX, Sitoinosides X.

5.1.4 Salts:
Cuscohygrine, Anahygrine, Tropine, Pseudotropine, Anaferine
5.1.5 Flavonoids:
Kaempferol, Quercetin.
5.2 Chemicals responsible for immunomodulatory effects
Till to date, various studies have been performed in order to find out the exact chemical
compounds which are responsible for immunomodulatory effects of withania somnifera plant
and it was concluded that Glycowithanolides, Sitoindosides IX & X, Withaferin A,
Withanolide E are the major chemical constituents responsible for its immunomodulatory
effects.

In a study, Bone evaluated Glycowithanolides and a mixture of Sitoindosides IX & X isolated

from W. somnifera for its immunomodulatory effects and concluded that at a dose of 50-
200mg/kg orally these constituents causes a significant movement and activation of peritoneal
macrophages, phagocytes and lysosomal enzymes.

In year 2003 Jayaprakasm et al studied the immunomodulatory effect of Withaferin-A and

suggest that it was observed that Withaferin-A showed an enhanced immunomodulatory
activity than doxorubicin in inhibiting growth of breast and colon cancer cell lines.

Aggarwal et al in 2012 suggested that Withaferin A and Withanolide E exhibited specific

immunosuppressive effect on human T and B lymphocytes and on mice thymocytes. They
further explained that withaferin A had non-specifically targeted both B & T lymphocytes
while Withanolide E had specifically targeted T lymphocytes.

A recent observational study has been performed in 2019 by Kumar et al on Balb/c Mice which
were intoxicated with Zinc Oxide nanoparticles along with W. somnifera extract and
Withanolide A for 28 days and it was found that the toxicity of zinc oxide nano particles has
been markedly reduced in the presence of W. somnifera extract and Withaferin A as they caused
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decrease in arginase gene expression and increased restoration of phagocytic activity of Balb/c
mice.

6 Doses
F. Malik et al in year 2007 performed a study to investigate the effectiveness and dose of a
chemically standardized aqueous alcoholic (1:1) root extract of W. somnifera (AGB) as
immunomodulator. In this study they investigated the effects of orally administered graded
doses of 10mg, 30mg, and 100 mg/kg (in 200μl of 1%gum acacia in saline) of standard extract
of W. somnifera on the immune system of SRBC immunized BALB/c mice for 15 days. At the
end of study they came with the finding that at a dose of 30mg/kg body weight W. somnifera
extract stimulated cell mediated immunity, increased IgM & IgG titers, enhanced proliferation
and differentiation of lymphocytes, it enhanced selectively type-1 immunity by increasing T
helper cell 1, cytokines interferon (IFN)-γ and interleukin (IL)-2 expression. So the study
suggested that a dose of 30mg/kg body weight of a standard extract of W. somnifera shows its
immunomodulatory effects.

7 Toxicity
The acute and sub-acute oral toxicity investigation of hydro-alcoholic root extract of W.
somnifera at a dose of 2,000 mg/kg/ body weight were done in Wistar rats, after administration
of extract animals were watched for 14 days in case of acute toxicity, while in case of sub-
acute toxicity the extract was administered at 500, 1,000, and 2,000 mg/kg doses and noticed
for 28 days and at the end of study author concluded that there were no any significant changes
were seen in the body and organ weight and haematological parameters in the rats and even the
extract was found to be safe from death, structural deformities and abnormal body growth.
Author also indicated that a daily dose of 1100mg/kg of W.somnifera extract intraperitoneal
injection in Swiss albino mice doesn’t cause any death in 24 h of injection but, a little increase
in dose causes death with an LD50 of 1,260 mg/kg/body weight in acute toxicity study. So,
from above study it was concluded that W. somnifera extract is relatively safer in all age groups
for the treatment of various diseases below LD50 i-e 1260mg/kg.

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8 References
1. F. Malik, e. a., 2007. A standardized root extract of Withania somnifera and its major
constituent withanolide-A elicit humoral and cell-mediated immune responses by up
regulation of Th1-dominant polarization in BALB/c mice. Life Sciences, pp. 1525-1538.

2. Jitendra Kumal, M. D. M. A. H. G. K., 2019. Exploration of immunomodulatory and protective

effect of Withaniasomnifera on trace metal oxide (zinc oxide nanoparticles) induced toxicity
in Balb/c mice. Molecular biology Reports, 07 March.pp. 1-13.

3. Leemol Davis, G. K., 1999. Immunomodulatory activity of Withania somnifera. Journal of

Ethnopharmacology, pp. 193-200.

4. Madhuri Sharma, G. P., 2014. Studies on phytochemistry and toxicity of Withania somnifera.
International Journal of Animal, Veterinary, Fishery and Allied Sciences, pp. 12-16.

5. Md. Khorshed Alam, M. O. H. a. M. S. U., 2016. Therapeutic use of Withania somnifera

(Review). Asain Journal of Medical and Biological Research, pp. 148-155.

6. Namdeo, D. S. M. (. &. A. G., 2020. Pharmacological evaluation of Ashwagandha highlighting

its healthcare claims, safety, and toxicity aspects. Journal of Dietary Supplements, 03 April.pp.
1-45.

7. Nasreen Jahan, S. M., 2016. Medicinal properties, phytochemistry and pharmacology of

Withania somnifera: an important drug of Unani Medicine. Journal of Scientific and Innovative
Research, 20 july.pp. 156-160.

8. Nawab John Dar, A. H. M. A., 2015. Pharmacologic overview of Withania somnifera, The Indian
Ginseng.. Cellular and Molecular Life Sciences, 26 August .pp. 1-16.

9. Neeraj Tandon, S. S. Y., 2020. Safety and clinical effectiveness of Withania Somnifera (Linn.)
Dunal root in. Journal of Ethnopharmacology, pp. 1-13.

10. Neha Singh, D. A. R. J. S., 2019. Biochemical and Therapeutic Properties of Withania Somnifera
in Traditional Medicinal system. International Journal of Trend in Scientific Research and
Development (IJTSRD), 3(3), pp. 551-556.

11. Ruchi Tiwari, S. C. M. S. K. D. s. V. S., 2014. Ashwagandha (W. somnifera): Role in safeguarding
Health, Immunomodulatory effects, Combating Infection and Therapeutic application: A
Review.. Journal of Biological Sciences, pp. 77-94.

12. Sumaira Saleem, G. M. M. A. H. M. A. S. N. A. B., 2020. Withania somnifera L.: Insights into the
phytochemical profile,therapeutic potential, clinical trials, and future prospective.. Iranian
Journal of Basic Medical Sciences, 21 June .pp. 1501-1526.

13. Swagat Kumar Mallick, U. M. G. M., 2020. HEALTH BENEFITS OF WITHANIA SOMNIFERA (L.)
DUNAL: A REVIEW. An International Bilingual Peer Reviewed Refreed Research Journal,
December, 10(40), pp. 115-117.

14. Verma, S. K., 2011. Therapeutic uses of Withania somnifera (Ashwagandha) with a note
onwithanolides and its pharmacological actions.. Asian Journal of Pharmaceutical and Clinical
Research, july.pp. 1-4.

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