Summary of Effects:

1.  Antimicrobial activity against  fungi Aspergillus flavus, Fusarium oxysporum, F. verticilloides and a range of bacteria including Salmonella typhimurium.

 3.Anti-inflammatory effects in animals including effectiveness against arthritis in animals

4. Antioxidant effects Animal studies have demonstrated withanolides inhibit cyclooxygenase enzymes, lipid peroxidation, and proliferation of tumor cells

5  Memory enhancement effects

Historical and Traditional Use:

 Ashwagandha is also alleged to be able to treat the following conditions:

·        Tumors

·        Cancer

·        Fevers

·        Inflammatory conditions

·        Arthritis

·        Asthma

·         Hypertension

·        Infections

·        Cholesterol-lowering

·        Cardioprotective

Three main effects have been extensively studied in animals, which include anti-cancer effects, anti-inflammatory effects and neurological/memory enhancing effects. There is also published material on its antimicrobial effects. Some animal studies also exist on the effects of ashwagandha on stress in animals and antioxidant effects. Human studies are lacking. Summaries of the main effects studied include:

1. Anti- stress:

 Researchers Archana and Nivasivayam from the Department of Physiology, University of Madras, India  stressed adult Wistar albino rats using a cold water swimming test and compared those pre-treated with Ashwaganda with untreated rats and found those pretreated with Ashwagandha showed fewer physiological signs of stress.( 2. Archana and Nivasivayam 1999)

Bhatnagar and colleagues found Ashwandha effective in reducing stress induced gastric ulcers  in rats  ( Bhatnagar et al 2005)

Dhuley demonstrated a reduction in the stress-induced rise in lipid peroxidation in experimental rabbits and mice treated with ashwagandha supporting its anti-stress properties.(.Dhuley 1998)

2. Anti-inflammatory Activity.

Rasool and Varalakshmi investigated the immunomodulatory effects of an aqueous suspension of the powdwered root of Ashwagandha both in vitro and in vivo. It showed potent inhibitory activity towards the complement system, mitogen induced lymphocyte proliferation and delayed-type hypersensitivity reaction. Administration of Ashwagandha root powder did not have a significant effect on humoral immune response in rats. Their results demonstrated immunosuppressive effects of Ashwagandha root powder, suggesting it could be a candidate for developing as an immunosuppressive drug for inflammatory diseases(  Rasool and Varalakshmi 2007).

 Researchers implanted cotton pellets in rat paws and assessed the inflammatory response with and without application of methanolic extract of the aerial parts of  ashwagandha. Findings demonstrated antiinflammatory activities comparable to that of hydrocortisone sodium succinate in Ashwagandha treated rats. ( al-Hindawi et al 1992)

Ashwaganda has also been shown effective in the treatment of experimentally-induced gouty arthritis in rats.{.Rasool and Varalakshmi 2006)

3. Anti-Cancer  Effects and Adjuvant Role in Cancer Treatment

From the many studies published it can reasonably it can be concluded that in animals ashwagandha reduces tumor cell proliferation while increasing overall animal survival time.

Devi and colleagues demonstrated a regression in sarcomas in rats treated with ashwagandha injections at doses of 500 to 750mg/ kg. (9 Devi 1992)  Prakash and colleages demonstrated a reduction in DMBA induced squamous cell skin cancers in mice when treated with maximally tolerated doses of Ashwaganda for 24 weeks.(.Prakash et al 2002)

Devi demonstrated that an alcoholic extract of the dried roots of Ashwagandha as well as withaferin A isolated from the extract showed significant antitumor and radiosensitizing effects  in experimental tumors inChinese hamsters in vivo, without any noticeable systemic toxicity. (.Devi 1996)

The administration of an extract of Withania somnifera was found to significantly reduce leucopenia induced by cyclophosphamide treated experimental animals. Administration of an extract from the powdered root of the plant Withania somnifera  enhanced the levels of Interferon gamma (IFN-gamma) (75.87 pg/ml), Interleukin-2 (IL-2) (14.16 pg/ml) and Granulocyte macrophage colony stimulating factor (GM-CSF) (49.22 pg/ml) in normal Balb/c mice. (12.Davis and Kuttan 1999) Administration of Withania somnifera extract (Solanaceae) was found to significantly reduce leucopenia(low white cell count) induced by cyclophosphamide (CTX) treatment. The total WBC count on the 12th day of the CTX-treated group was 3720 cells/mm3 and that of CTX along with Withania was 6120 cells/mm3. Treatment of Withania along with CTX was found to significantly (P < 0.001) increase the bone marrow cellularity (13.1 x 10(6) cells/femur) compared to CTX alone treated group (8 x 10(6) cells/femur)(13. Davis and Kuttan 1998). In a further study the administration of Ashwagandha(20 mg/dose/animal) for five days in conjunction with cycloposphamide was associated with a reduction in urotoxicity.( 14. Davis and Kuttan 2000)

 The steroidal lactone withaferin A displayed significant antitumor and radiosensitizing effects, inhibiting tumor growth and increasing survival in Swiss mice inoculated with Ehrlich ascites carcinoma (14.Devi et al 1995; 15.Sharad et al 1996). The administration of methanolic extract of Ashvagandha was found to significantly increase the WBC count in normal Balb/c mice and reduce leucopenia induced by a sublethal dose of gamma radiation. ), Interleukin-2 (IL-2) (14.16 pg/ml) and Granulocyte macrophage colony stimulating factor (.Kuttan 1996)

Ichikawa and colleagues investigated the anti-carcinogenic effects in animal and cell cultures by decreasing the expression of nuclear factor-kappaB, suppressing intercellular tumor necrosis factor, and potentiating apoptotic signalling in cancerous cell lines.

Padmavathi and colleagues studied the protective effects of Ashwaganda in experimentally induced skin and stomach tumours in mice. They demonstrated Ashwagandha roots inhibited phase I and activated phase II and antioxidant enzymes in the liver. Further, in a long-term tumorigenesis study, Withania inhibited benzo(a)pyrene-induced forestomach papillomagenesis, showing up to 60 and 92% inhibition in tumor incidence and multiplicity, respectively. Similarly, Withania inhibited 7,12-dimethylbenzanthracene-induced skin papillomagenesis, showing up to 45 and 71% inhibition in tumor incidence and multiplicity. In both studies, Withania showed no apparent toxic effects in mice as monitored by the body weight gain profile.( Padmavathi et al 2005)

Ashwagandha treatment significantly downregulated the gene and protein expression of proinflammatory cytokines IL-6, IL-1β, chemokine IL-8, Hsp70 and STAT-2, while a reciprocal upregulation was observed in gene and protein expression of p38 MAPK, PI3K, caspase 6, Cyclin D and c-myc. Furthermore, Ashwagandha treatment significantly modulated the JAK-STAT pathway which regulates both the apoptosis process as well as the MAP kinase signaling. These studies outline several functionally important classes of genes, which are associated with immune response, signal transduction, cell signaling, transcriptional regulation, apoptosis and cell cycle regulation and provide insight into the molecular signaling mechanisms that are modulated by Ashwagandha, thereby highlighting the use of this bioflavanoid as effective chemopreventive agent relevant to prostate cancer progression.

Simultaneous administration of Withania extract (122 +/- 10 tumour nodules) and Withanolide (126 +/- 9 lung tumour nodules) could significantly (p < 0.001) inhibit the metastatic colony formation of the melanoma in lungs of mice..(20. Leyon and Kuttan)

 Ashwagandha root extract enhanced the levels of Interferon gamma (IFN-gamma) (75.87 pg/ml), Interleukin-2 (IL-2) (14.16 pg/ml) and Granulocyte macrophage colony stimulating factor (GM-CSF) (49.22 pg/ml) in normal Balb/c mice. (12.Davis and Kuttan 1999)

 Ashwagandha has also been studied as an adjuvant agent in minimizing adverse effects associated with anti-cancer agent cyclophosphamide:.

 Administration of Ashwagandha extract (Solanaceae) was found to significantly reduce leucopenia(low white cell count) induced by cyclophosphamide (CTX) treatment. The total WBC count on the 12th day of the CTX-treated group was 3720 cells/mm3 and that of CTX along with Ashwagandha was 6120 cells/mm3. Treatment of Withania along with CTX was found to significantly (P < 0.001) increase the bone marrow cellularity (13.1 x 10(6) cells/femur) compared to CTX alone treated group (8 x 10(6) cells/femur)(13. Davis and Kuttan 1998). Its use as an adjuvant treatment to prevent radiotherapy associated adverse effects was further studied by Kuttan.

Administration of a 75% methanolic extract of the plant was found to significantly increase the total WBC count in normal Balb/c mice and reduce the leucopenia induced by sublethal dose of gamma radiation. Treatment with Ashwagandha was found to increase the bone marrow cellularity significantly, the percentage increase being 146.3. Treatment with Ashwagandha had normalised the ratio of normochromatic erythrocytes and polychromatic erythrocytes in mice after the radiation exposure. Major activity of ashwagandha.  seemed to be in the stimulation of stem cell proliferation.(Kuttan 1996) In a further study the administration of Ashwagandha(20 mg/dose/animal) for five days in conjunction with cycloposphamide was associated with a reduction in urotoxicity.( 14. Davis and Kuttan 2000)

Antioxidant

An aqueous suspension of root extract of Ashvagandha prevented the rise of experimentally induced lipid peroxidation in rabbits and mice (19. Dhuley 1998). An extract of ashwagandha produced an increase in the levels of superoxide dismutase, catalase and glutathione peroxidase in rat brain, all of  which are potent antioxidant enzymes in the human body. This supports reports that it is an antioxidant with immunomodulant and antiinflammatory activity (20. Bhattacharya et al 1997).

A glucose tolerance test  performed in the group treated with Ashwaganda, showed a significant improvement in glucose tolerance.  Insulin sensitivity was assessed for both peripheral insulin resistance and hepatic insulin resistance. Ashwagandh treatment significantly improved insulin sensitivity index (K(ITT)) that was significantly decreased in NIDDM control rats. There was significant rise in homeostasis model assessment of insulin resistance (HOMA-R) in NIDDM control rats whereas WS treatment significantly prevented the rise in HOMA-R in NIDDM-treated rats. Our data suggest that aqueous extract of WS normalizes hyperglycemia in NIDDM rats by improving insulin sensitivity (27. Anwer et al 2008)

Historically the roots and berries are used medicinally. The fresh roots may be boiled in milk prior to drying in order to leach out any undesirable constituents. Recommended dose is 3–6 grams of the dried root, taken each day in capsule or tea form. To prepare a tea, 3/4–1 1/4 teaspoons (3–6 grams) of ashwagandha root are boiled for 15 minutes and cooled; 3 cups (750 ml) may be drunk daily. Alternatively, tincture 1/2–3/4 teaspoon (2–4 ml) three times per day, is sometimes recommended  The berries are used in cheese-making to coagulate milk.

21.The Tumor Proteasome Is a Primary Target for the Natural Anticancer Compound Withaferin A Isolated from "Indian Winter Cherry"