Chapter 3

REVIEW OF LITERATURE

13 | P a g e
Literature review was focused on the developments in the following four aspects of RA.

1. RA animal models

2. Recent understanding of pathogenesis of RA

3. Current trends in the therapy

4. Medicinal plants evaluated for their effects in animal models of RA.

3.1 RA animal models:

Animal models are developed to evaluate the potential therapeutic drugs as well as understand

the pathogenesis of RA. The choice of the animal model depends on the following criteria (25).

1. Animal model should have morphological similarities with human RA

2. Should be able to assess the efficacy of various drugs.

3. Model should be easily developed

4. Data should be reproducible

5. The duration of the assessment period should be reasonable

Many models have been developed to study the anti- rheumatoid activity. At present there is no

ideal model that produces typical RA as seen in humans. However genetic animal models

develop RA closer to that in humans.

Currently used models are:

14 | P a g e
Rat models

1. Collagen (Type II) induced arthritis (26)

2. Fruend’s complete adjuvant induced arthritis (27)

3. Antigen induced arthritis (28)

4. Streptococcal cell wall induced arthritis (29)

5. Pristane-induced arthritis (30)

6. Carrageenan- induced joint inflammation (31)

7. Zymosan induced arthritis (32)

8. Kaolin induced arthritis (33)

9. Ankle joint urate arthritis (34)

Mouse models

1. Collagen (Type II)-induced arthritis (35)

2. Pristane-induced arthritis (36)

3. Proteoglycan-induced arthritis (37)

4. Zymosan-induced arthritis (38)

5. Immune complex arthritis (39)

6. G6PI-induced arthritis (40)

Genetic models

1. K/BxN (41)

2. HuTNF Tg (42)

3. Human TNF–transgenic mice (43)

15 | P a g e
Among these models RA induced in Rats/ Mice with FCA and type II collagen are considered

the most economic and appropriate models to assess the efficacy and safety of newer agents to

generate reproducible preclinical data.

As there are no reported differences in the immuno pathogenesis of arthritis induced in both the

models. FCA induced RA model is selected for the current study (44).

Freund's complete adjuvant induced arthritis in rats

It was first described by Newbold BB in 1963. FCA contains 10 mg of mycobacterium

tuberculosis bacilli suspended in 10 ml of heavy paraffin oil. FCA is found to release PGE2,

TNF-α and nitric oxide and increase the level of myeloperoxidase contributing to the

development of arthritis. It induces acute inflammatory reaction as well as immunological

changes approximately in 9 days. This model is sensitive to anti-inflammatory and immune

suppressive agents and considered relevant for the study of patho-physiological and

pharmacological control of inflammatory process as well as for the evaluation of analgesic drugs

(45).

Procedure

In this model, 0.1 ml of FCA is injected subcutaneously into the left hind paw of rats on zero

day. Clinical evidence of arthritis occurs from day 8-9 with swelling of hind paw (onset of

disease). After about 12 days it becomes difficult for the rats to move due to paw swelling. Drug

therapy is started either on day 0 (prophylactic model) or day 9 (therapeutic model) (25).

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3.2 Recent understanding of pathogenesis of RA

Faulty immune response is identified as the cause for RA. The complex interaction between

environmental and genetic factors induces an antigenic response stimulating the CD4+ T cells

present in the synovium. Activation of CD4 T cells results in two major responses 1) activation

of B cells resulting in the production of plasma cells which release immunoglobulins such as RF

and anti-CCP. 2) Differentiation of CD4 Th cells into Th-1 and Th-2. Another type of Th cell,

Th-17 has been identified recently which is found to play an important role in the pathogenesis

of RA. Inflammatory mediators released by Th-1, Th-2 and Th-17 have been given in the Figure-

1. All these pro-inflammatory cytokines and inflammatory mediators initiate the inflammatory

response in the joint resulting in neutrophil infiltration, macrophage and fibroblast activation (46,

47).

TNF-α is the main inflammatory inducer that drives pathogenesis and development of RA. It has

been clinically confirmed that TNF blockade can effectively control the severity of RA.

Activated T cells and macrophages can produce not only TNF- α but also IL-1, IL-6 and IL-17,

which cause chronic inflammation in the synovium by different mechanisms (48). TNF-α can

degrade the articular cartilage and bone by stimulating osteoclast activity and the activity of

enzymes such as MMP, leading to joint deformity. RANKL which is a target for TNF is found to

be highly expressed in RA and is associated with the bone loss (49). Additionally, the

rheumatoid factor released by B cells may form an immune complex with self-antigens and

trigger inflammation through activation of the complement system. Similarly antibodies

developed against citrulline also stimulate T-cells and initiate inflammatory process (20). The

17 | P a g e
abnormality in the normal programmed cell death may be also involved in joint inflammation in

RA. Thus multiple mechanisms are involved in the pathogenesis of RA.

Role of different cytokines in RA (19):

TNF-ɑ causes :

 Activation of monocytes, release of PG

 Increased WBC and granulocytes priming

 Apoptosis of T-cell , TCR dysfunction

 Increased expression of endothelial cell adhesion molecules

 Decreased collagen synthesis

 Increased matrix metalloproteinase

IL-1:

 Increase in the release of synovial fibroblast and monocyte cytokine, chemokine, MMP

and PG

 Production of reactive oxygen intermediate

 Activation of osteoclasts

 Expression of endothelial cell adhesion molecules

IL-6:

 Activation of osteoclasts

 Recruitment of neutrophils

18 | P a g e
  VEGF production

 Formation of pannus

 Proliferation of B-cell and production of antibodies

 Proliferation of T-cell and differentiation

IL-17:

 Infiltration and activation of T-cells

 Monocytes, fibroblasts and neutrophils recruitment

 Increased production of chemokines at the site of inflammation

 Increased IL-6 and immune response

 Osteoclastogenesis and cartilage destruction

 Integration of IL-1b, TNF-a and IFN-γ activities

 VEGF production, angiogenesis and pannus formation

All these mediators contribute to the progressive pathological changes that occur in RA

19 | P a g e
 Inflammatory pathway in Rheumatoid arthritis

Genetic predisposition Triggering factors

Synovial CD 4 T cell activation

B cells activation Differentiation to Th1 and Th2

IgM, RF, Anti-CCP Ab

Th1→ IFN-γ, TNF-α, Th-2→ IL-4, 5, 6, 10 &

13

Th17- IL-6, 17, 21, 22, 26,

TNF α & GMCSF

Macrophages- IL 1, IL 21, IL 23
Synovial fibroblast- IL 18, RANKL

Release of inflammatory mediators like PGs,

Inflammation with neutrophil infiltration into
the joint - release of MMP, cartilage damage

Figure-1: The pathogenesis of RA is schematically given above.

Thus, controlling the key cellular and humoral immune responses and the dominant pro-

inflammatory cytokine production may be the possible therapeutic targets for treatment of RA.

20 | P a g e
3. 3 Current trends in the therapy

The treatment is aimed at targeting these mediators relieving pain, improving mobility and

arresting articular damage. Anti-inflammatory analgesics and immunomodulators are the main

groups of drugs used and these include NSAIDs, DMARDs and Immunosuppressives.

NSAIDs & steroids are commonly used initially in the treatment of RA, Both act at different

stages in inflammatory pathway.

PGs are the main inflammatory mediators. The production of PGs from arachidonic acid is

mediated by the enzyme cyclo-oxygenase (COX). NSAIDs inhibit COX enzyme there by the

production of PGs. COX exists in two isomeric forms COX-1 and COX-2. COX-1 is a

constitutive enzyme present in all normal tissues especially platelets, stomach and kidney. COX-

2 is produced at the site of and during inflammation. COX-1 is mediates the release of PGs in the

stomach which causes mucus production, improves blood flow and thus offers protection against

gastric ulceration. In the kidney prostaglandin production by COX-1 maintains renal blood flow

and in platelets inhibits their aggregation. When non selective COX inhibitors such as

indomethacin, ibuprofen and diclofenac are used, though they offer relief from joint swelling and

pain, they cause adverse effects such as peptic ulcer, nephro-toxicity and bleeding diathesis as a

result of COX-1 inhibition. Hence selective inhibitors of COX-2 such as rofecoxib, valdecoxib

which act only at site of inflammation were introduced so that these adverse effects can be

avoided but COX-2 inhibitors caused lethal cardiac toxicity hence withdrawn from use (50).

Because of the adverse effects of NSAIDs, glucocorticoids like predinisolone and methyl

prednisolone are used either alone or along with NSAIDs so that the dose of both the drugs can

be minimized. Glucocorticoids act at a higher level in the inflammatory process by inhibiting the

21 | P a g e
production of arachidonic acid from cell membrane phospho lipid. The immunosuppressive

effects of glucocorticoids further help the treatment of RA. But unfortunately long term use of

glucocorticoids leads to several systemic side effects such as peptic ulcer, hypertension, immune

dysfunction and osteoporosis (51). Hence DMARDs was introduced.

DMARDs are methotrexate, azathioprine, sulfasalazine, hydroxychloroquine, gold salts, D-

penicillamine and leflunomide. These agents decrease the inflammatory process and its

consequent destructive changes in RA by their immuno-modulatory effects. But long term use of

DMARDs is known to produce myelosuppression, stomatitis, alopecia, diarrhoea and

thrombocytopenia (50).

The next groups of drugs used are biologic DMARDs, which include etanercept, adalimumab,

infliximab, certolizumab and golimumab. They act by inhibiting TNF, one of the main

inflammatory mediators in RA (52).

The drugs which inhibit cytokines like IL-1, 6, TNF-α inhibit the initiation of inflammation.

Drugs which inhibit the cytokines are the biologicals. There are two groups of cytokines. One

which facilitates the inflammatory process is called pro-inflammatory cytokines and others

which inhibit inflammation are the anti-inflammatory cytokines. Both together maintain

homeostasis. When the activity of pro-inflammatory cytokines is increased inflammation occurs.

Hence the biologicals are used to inhibit pro-inflammatory cytokines or facilitate the production

of anti-inflammatory cytokines.

The first direct and selective IL-1 receptor antagonist, anakinra is used alone or in combination

with any of the DMARDs, except etanercept and infliximab because of the risk of development

of neutropenia. In addition to cytokine modulators drugs are also produced which can act against

22 | P a g e
other proteins involved in co-stimulation in inflammation such as cytotoxic T lymphocyte

associated protein (CTLA4), monoclonal antibodies against CD4 lymphocytes as well as IL

receptor proteins.

For example abatacept is a CTLA 4 fusion protein and is a selective co-stimulation modulator

which inhibits T-cell activation.

Similarly rituximab is anti CD 20 antibody; toclilizumab is anti IL-6 antibody and tofacitinib

Janus kinase inhibitor are often combined with methotrexate or other DMARDs to improve

efficacy (53).

All these biological DMARDs can also cause adverse effects such as hypotension, severe

infections, organ toxicity and others.

Thus the treatment of RA is incomplete and one has to use many drugs to control the disease

process at different levels. Multiple drug therapy can cause multiple organ toxicity and other

long term side effects.

23 | P a g e
3.4 Medicinal plants evaluated for their effects in animal models of RA.

Among the plants screened for their effects in RA, 10 plants used in FCA induced arthritis have

been selected for review.

1. Calotropis procera

V. L. Kumar and S. Roy had studied the effect of methanolic extract of Calotropis procera in

FCA induced arthritis. FCA caused the release of PGE2, TNF-α and nitric oxide and increased

the level of myeloperoxidase contributing to the development of arthritis. Calotropis procera

extract of 500 mg/kg produced marked reduction in the inflammatory mediators and response to

the same extent as that of standard drug rofecoxib (100 mg/kg) (54).

2. Hemidesmus indicus

The anti-arthritic activity of four different extracts, hydroalcoholic extract (450 mg/kg)

chloroform extract (60 mg/kg), ethyl acetate (75 mg/kg) and residual fraction (270 mg/kg) of

Hemidesmus indicus root in adjuvant induced arthritis in rats was evaluated by Mehta A. The

ethyl acetate extract was found to be better than other fractions and it contained the phyto-

chemicals terpenes, sterols and phenolic compounds which could have contributed to the anti-

arthritic activity (55).

3. Tridax procumbens

The anti-arthritic activity of T. procumbens was studied in FCA induced arthritis in rats in the

doses of 250 and 500 mg/kg. This study had shown dose dependant anti-arthritic effect is

24 | P a g e
comparable to that of Indomethacin. T. procumbens in addition inhibited the increase in liver

markers induced by FCA (56).

4. Gymnema sylvestre

The anti arthritic effects of petroleum ether extract (300 mg/kg) and aqueous extract (300 mg/kg)

of Gymnema sylvestre were evaluated in FCA induced arthritis in rats. Paw volume and blood

parameters, TC DC, ESR, Hb were measured. The authors have concluded that both the extracts

of Gymnema sylvestre exhibited better anti-arthritic activity compared to untreated group.

Tannins and saponins are the main phytochemicals present in Gymnema sylvestre and are known

to possess anti-arthritic activity (57).

5. Syzygium cumini

The anti-arthritc activity of methanolic extract of Syzygium cumini seeds at the doses of 250

mg/kg and 500 mg/kg on FCA induced arthritis was evaluated by Eshwar kumar et.al. Treatment

Syzygium cumini showed significant effect in preventing PV and improved RBC count, Hb level

and ESR to normal level and significantly decreased the WBC count when compared to

untreated RA group. The effect was found to be equal to standard drug indomethacin (58).

6. Terminalia chebula

Ramani YR et.al, have reported the anti-arthritic activity of acetone extract of fruits of

Terminalia chebula in FCA induced arthritis in Wistar rats. Terminalia chebula at the doses of

160 and 320 mg/kg showed good reduction in ESR, paw edema and joint thickness which is

comparable to dexamethasone treated group. Terminalia chebula produced significant anti-

25 | P a g e
inflammatory and anti-arthritic activities at the dose of 320 mg/kg and the mechanism attributed

was inhibition of hyaluronidase and collagenase, preventing the degradation of articular cartilage

(59).

7. Ficus bengalensis

The methanolic extract of Ficus bengalensis bark was evaluated in three doses of 100, 200 and

300 mg/kg (i.p) in FCA induced arthritis, Formalin induced arthritis and Agar induced arthritis in

rats by Manocha N et.al. The extract had shown anti-rheumatic activity in a dose dependent

manner in all the models (60).

8. Vitex negundo. Linn

Cheng-Jian Zhenga et.al, screened the effects of Vitex negundo. Linn seeds extract (EVNS) on

FCA induced arthritis in rats. EVNS in 2 doses- 340 mg/kg and 85 mg/kg had significantly

inhibited the PV, reduced the arthritis score and decreased the inflammatory markers. EVNS had

potential therapeutic benefit in arthritis and the mechanism attributed was by reducing TNF-α,

IL-1β and IL-6 levels and elevating IL-10 in serum and in addition down-regulation of COX-2 &

5-LOX (61).

9. Strychnos potatorum Linn

Sanmugapriya Ekambaram et.al evaluated the effects of the aqueous extract and the whole seed

powder of Strychnos potatorum Linn seeds on the FCA induced arthritis in rat. The group treated

with extracts, showed significant reduction in PV. Hb, RBC, WBC, ESR, blood urea, serum

creatinine, total proteins and acute phase proteins were significantly brought back to near normal

26 | P a g e
level by the extract at the dose of 200 mg/kg/p.o. Further the histopathological and X-ray studies

confirmed the anti-arthritic activity of Strychnos potatorum (62).

10. Ncytanthes arbor-tristis

Brijesh Rathore et.al evaluated the impact of long term use of aqueous and ethanolic extracts of

the leaves, fruits and seeds of Nyctanthes arbor-tristis (NAT) on the modulatory effect of pro-

and anti-inflammatory cytokines in FCA induced arthritis in mouse. It was found that treatment

with extracts from leaves and fruits of NAT at the dose of 25 mg/kg had reduced the TNF-α, IL-

1β and IL-6 from day 14 while seed extracts were ineffective, suggesting that the active

component is in the hard covering of the fruit (63).

From the literature review it is inferred that

A) FCA induced arthritis in rats can be selected as the appropriate economic model for

evaluating potential therapeutic agents for RA.

B) Pro-inflammatory cytokines IL-1, IL-5, IL-6, IL-17, IL-21, IL-22 and IL-26, TNF-α, GMCSF

& IFN-γ and anti-inflammatory cytokines such as IL-4, IL-10, IL-11 and IL-13 are in

homeostasis to prevent inflammation in RA. Hence agents that facilitate the release and activity

of anti-inflammatory cytokines and which inhibit pro-inflammatory cytokines will be useful in

the treatment of RA.

C) Several medicinal plants have been studied for their effects in RA. In these studies FCA

induced arthritis model has been mainly used. The evaluated plants are all found to be effective

in FCA induced RA. These plants are individually evaluated but not in combination and they are

not domestically used plants in day to day life.

27 | P a g e
Hence the two plants OS and SI which are used domestically in day to day life, readily available,

easy to collect and less expensive have been selected for evaluation in FCA induced arthritis in

rats with the objectives of assessing their safety and efficacy individually and in combination as

well as their mechanism of action in comparison with standard drugs diclofenac and

methotrexate.

28 | P a g e
3.5 Ocimum sanctum
Introduction of plant:

Ocimum sanctum L (Holy basil) is one among the 60 species of the genus Ocimum and belongs

to the family Lamiaceae. It is indigenous to the tropical regions of Asia and America. OS is

wildly cultivated in India. A wide range of medicinal properties have been attributed to OS and

different parts of the plants such as leaves, flowers, stems, roots and seeds are extensively used

in Indian systems of medicine for different disorders such as fever, common cold, sore throat,

headache, eye diseases, dental and skin disorders, insect bites, renal stones and stress disorders

(64).

The different species of OS are used in food, drug development and perfumary (65). Holy basil

has also been described as Ocimum tenuiflorum or Ocimum gratissimum (66). OS is the most

commonly used name due to the wide range of its uses in religious and cultural traditions. It is

closely related to sweet basil (Ocimum basilicum), a commonly used herb in Europe and North

America.

OS is locally known as Tulsi. Three types of Tulsi are in use; Tulsi with green leaves is Sri

Tulsi or Rama Tulsi and is the commonest type. The plant with dark green to-purple leaves is

known as Krishna Tulsi and the third one is a wild variety called Vana Tulsi that often grows in

the forest.

Of these, the Rama and Krishna holy basil are most commonly grown in homes as well as

commercially produced for use in ayurvedic preparations. The plant is found mainly in

subtropical and tropical areas of Asia including India, China, Malaysia, Sri Lanka, and Thailand

29 | P a g e
in addition to Australia and Africa, at altitudes of up to 1800 meter in India to sandy dry

conditions in China (67).

Morphology

OS is an erect shrub, with many branches and growing to about 30-60 cm height with green or

purple leaves placed opposite on hairy stems. Leaves are about 5 cm long, usually somewhat

toothed. Flowers are purple in color in elongate racemes arranged in whorls.

Taxonomy of Ocimum sanctum

Kingdom Plantae

Division Magnoliophyta

Class Magnoliopsida

Unranked Asterids

Order Lamiales

Family Lamiaceae

Genus Ocimum

Species O. sanctum

Binomial name Ocimum sanctum L.

30 | P a g e
Vernacular names of Ocimum sanctum

English Holy basil

Hindi Tulsi

Sanskrit Tulasi, Surasa, Gramya

Tamil Thulasi

Kannada Tulasi

Telugu Tulasi

Malayalam Trittavu

Images of Ocimum sanctum

A) B) C)

Image-1: A) Rama tulsi, B) Krishna tulsi and C) Vana tulsi

Source: http://www.astrogle.com/images/2015/08/tulasi_holy_basil.jpg Accessed on 20.05.2016

http://www.khichdionline.com/wp-content/uploads/2015/04/136.jpg Accessed on 20.05.2016

http://www.herbalremediesadvice.org/images/Vana-Tulsi.jpg Accessed on 20.05.2016

31 | P a g e
Phytochemicals present in OS

Os contains many nutrients and active biological compounds, the ratio of which will be

different among different strains and even among strains cultivated in same area which may be

due to variation in the way of growing, harvesting, processing and storage (68).

OS is found to have many phytochemicals such as tannins, saponins, flavonoids, steroids,

terpenoids, cardiac glycerides, eugenol, euginal (also called eugenic acid), ursolic acid,

rosmarinic acid, carvacrol, linalool, limatrol, caryophyllene, methyl carvicol, cyclohexane,

cyclopentane, benzene methanamine and octadecane, palmitic acid, stearic acid, oleic acid,

linoleic acid and α –linolenic acid (69, 70).

Nutritional content

Vitamin C and vitamin A, calcium, zinc and iron, chlorophyll and many other nutrients are

present in OS. The Protein content is 4.2 g; carbohydrate 2.3 g; fat: 0.5 g; calcium: 25 mg;

phosphorus 287 mg; iron: 15.1 mg and edible portion contains 25 mg vitamin C per 100 g (71).

Pharmacological actions of OS

Anti-inflammatory and analgesic activity: Ethanolic extract of leaves of OS was evaluated

for anti-inflammatory activity in carrageenan induced paw edema in rats and analgesic activity

using hot plate method in mice by Hannan, J. M. A., et al. and they have reported that OS at the

dose of 250 mg/kg and 500 mg/kg had caused significant reduction of PV and an increase in

reaction time to pain establishing its anti-inflammatory and analgesic activity (72).

32 | P a g e
Antioxidant and wound healing: Shetty et.al. had evaluated the in-vitro antioxidant & wound

healing activity of ethanolic and aqueous extracts of OS leaves in Wistar albino rats. 400 and

800 mg/kg of both the extracts had shown increased levels of hydroxyproline, hexuronic acid,

hexosamines & wound breaking strength and decreased percentage of wound contraction. The

antioxidants SOD, catalase, GSH levels had been significantly elevated and LPO level was

reduced in treated group. Aqueous extract of 800 mg/kg had shown greater effect compared to

ethanolic extract. OS leaves extracts was found to contain phenolic compounds which had

greater free radical scavenging activity against different ROS (73).

Anticancer activity: Shivpuje P et.al evaluated the anti-proliferative effect and cytotoxic

activity of aqueous extract of OS leaves on KB mouth epidermal carcinoma cell line by MTT

assay. The extract had significant cytotoxic effect against oral cancer cell line. (74)

Diuretic effect: The diuretic effect of ethanolic extract of OS leaves was studied in animal

model by Preethi G Pai. 250 mg/kg and 500 mg/kg of the extract had shown increased diuretic

index to 1.65 and 2.26 respectively and the saluretic index as reflected by the Na/K ratio to 2.2

at 250 mg/kg and 2 at 500 mg/kg which was greater than that of furosemide which showed a

saluretic index of 1.81 (75).

Anti-diabetic activity: Anti-diabetic effect of ethanolic extract of OS in glucose fed and

streptozotocin treated rats was reported by Chattopadhyay RR. OS treatment resulted in

significant reduction in blood sugar in diabetic rats and enhanced the action of exogenously

administered insulin in control rats (76).

Anti-fatigue activity: The anti-fatigue activity of alcoholic extract of OS leaves in rats was

evaluated by Prasad et.al. Fatigue was induced by subjecting the rats to weight loaded forced

33 | P a g e
swim test every alternate day for 2 weeks. 150, 300 and 450 mg/kg of OS extract were

administered orally. Blood parameters like MDA and LA, glycogen, Hb, BUN and CK levels

were measured. The OS treatment resulted in significant increase in swimming time from day 1

to 14. MDA and LA levels in liver and muscle tissues, BUN and CK activity were significantly

reduced. 300 mg/ kg had showed better performance against fatigue than other two doses (77).

Antimicrobial activity: Vasudevan, D. M., et al evaluated the antimicrobial effect of alcoholic

and aqueous extracts of OS leaves. Aqueous extract was found to be a better inhibitor of gut

bacterial and fungal species than alcoholic extract. In addition the alcoholic extract showed

broader zone of inhibition for vibrioe cholera especially at a higher dose of 60 mg (78).

Cardio-protective activity: The effect of hydroalcoholic extract of OS leaves in isoproterenol

induced Myocardial infarction (MI) in rats was evaluated by Sharma M et.al. 25, 50, 75 & 100

mg/kg of OS significantly reduced the glutathione, SOD, LDH levels and inhibited lipid

peroxidation. 50 mg/kg produced greater cardio-protective effect (79).

Gastroprotective activity: Antiulcer effect of aqueous extract of OS against methanol induced

gastric ulcer in rats was reported by Dharmani P. OS at the dose of 100 mg/kg and 200 mg/kg

had 33.07% and 52.52% protection whereas omeprazole showed 60% protection against

methanol induced ulcers (80).

Anti-cataleptic activity: Pemminati Sudhakar, et.al evaluated the effect of ethanolic extract of

OS leaves on catalepsy induced by haloperidol. 1.75 mg/kg and 4.25 mg/kg body weight of OS

had better protection against catalepsy when compared to scopolamine & ondansetron in acute

as well as in chronic study (81).

34 | P a g e
Immunomodulatory activity: The immunomodulatory activity of aqueous extract of OS in rat

was reported by Jeba CR, et.al. Antibody titration, passive haemagglutination and chromic

chloride assays were performed. 100 & 200 mg/kg of OS extract produced significant elevation

in antibody titre and increased the RBC, WBC and Hb level (82).

Thus OS is found to have anti-inflammatory, analgesic, antioxidant, wound healing, anticancer,

diuretic, anti-diabetic, anti-fatigue, antimicrobial, cardio-protective, gastro-protective, anti-

cataleptic and immunomodulatory activities.

35 | P a g e
3.6 Sesamum indicum
Sesamum indicum (Sesame) belongs to the family, Pedaliaceae and one of the first known plants

used for its seeds. It has been used since ancient times as spice and is still an oil seed of global

significance. Its seed oil has a rich content of protein and is used in cooking and margarine

production (83).

Morphology

Sesame is an erect usually annual shrub reaches a height of 1.5 meter, depending on the type

and cultivating conditions. Some species are highly branched, whereas others are un-branched.

Seeds are black, brown or white, 2.5–3 mm long and about 1.5 mm wide. It is found in tropical,

subtropical, and southern temperate areas of the world, particularly India, China, South America

and Africa.

Taxonomy of Sesamum indicum

Kingdom Plantae

Unranked Angiosperms

Unranked Eudicots

Unranked Asterids

Order Lamiales

Family Pedaliaceae

Genus Sesamum

Species S. indicum

Binomial name Sesamum indicum L.

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Vernacular names of Sesamum indicum

English Gingelly, Sesame

Hindi Til, Tila

Sanskrit Tila

Tamil Ellu

Kannada Ellu

Telugu Nuvvulu

Malayalam Ellu

Images of Sesamum indicum

A) B) C)

Image-2: A) Sesame plant, B) White sesame seeds and C) Black sesame seeds

Source: http://www2.hcmuaf.edu.vn/data/phamductoan/image/Cayme/sesame.jpg Accessed on 20.05.2016

http://waynesword.palomar.edu/images2/sesame2b.jpg Accessed on 20.05.2016

37 | P a g e
Chemical composition of SI

SI seed extract is found to contain phytochemicals such as saponins, flavonoids, tannins,

glycosides and terpenes, sesamol, sesamin and sesamolin, oleic acid, α-tocopherol, γ-

tocophereol, palmitic acid, stearic acid and linoleic acid, α –linolenic acid (84, 85, 86).

Nutritional value

SI contains vitamin B1: 0.28 mg, 1.48 mg of copper, 0.88 mg of manganese, 120 mg of

tryptophan, 351.00 mg of calcium, 126.36 mg of magnesium, 5.24 mg of iron, 226.44 mg of

phosphorus, 2.80 mg of zinc and dietary fiber (87).

Pharmacological activities

Analgesic and antioxidant activity: Was evaluated by Nahar, L. 250 and 500 mg/kg of

alcoholic seed extract had produced 48.19% and 75.46% writhing inhibition respectively and

was found to be similar to ibuprofen which produced 71.82% inhibition with 25 mg/ kg.

SI had produced maximum antioxidant effect at the concentration of 120 µg/ml which was 92%

compared with standard antioxidant, ascorbic acid had produced 56% (88).

Hepato-protective activity: The hepato-protective effect of alcoholic extract of SI seeds in

carbon tetrachloride (CCL4) induced liver damage was studied by Kumar et.al. 400 mg/kg and

700 mg/kg of extract had reduced the protein synthesis and TG production and histological

changes in liver (89).

Antimicrobial activity: The antimicrobial effect of the silver nano-particles produced using SI

leaf extract against multi-drug resistant Escherichia coli (E. coli) had been evaluated by

Bokaeian M. The silver nano particle of SI leaf extract (400 mg/ml) inhibited all the micro-

38 | P a g e
organisms. Maximum inhibition was seen with E. coli followed by S. aureus and K. pneumonia

and no inhibition was observed for S. aureus. The least zone of inhibition was observed against

S. typhii. The highest and lowest MIC values against five isolates of E. coli were found to be

200 ppm and 50 ppm, respectively (90).

Anticonvulsant activity: Anticonvulsant activity of SI oil against experimental seizures in

mice was reported by Uma Advani. 10 ml/kg of S. indicum oil reduced the duration of MES

produced tonic hind limb extension. S. indicum oil was effective against PTZ and lithium +

pilocarpine induced seizures (91).

Antiobesity activity: Chinnala KM et.al evaluated the anti-obesity effect of methanolic extract

of SI in obesity induced rats using high fat diet. The extract of SI produced significant reduction

in BW, food intake, blood glucose level, protein, total cholesterol, LDL, VLDL, TG and an

increase in HDL level (92).

Nephro-protective activity: The nephro-protective activity of alcoholic extract of SI on renal

function in rats induced with diabetes by administration of combination of Streptozotocin and

Nicotinamide was reported by Bhuvaneswari P. SI significantly increased the serum total

protein, albumin and globulin and decreased the levels of urea, creatinine, uric acid and

improved the renal histological damage (93).

Anti cancer activity: Noor A et.al investigated the cytotoxic activity on Hep-2, AMN-3, RD

cell lines and one normal embryo rat cell line. The cytotoxic activity was observed at

concentration of 1000 μg/ml after 72 h. The percentage of cellular damage was 85.83%, 40.06%,

and 20.20% respectively in the above three cell lines (94).

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Fertility action: The ethanolic extract of SI and vitamin C in promoting fertility in male rats was

reported by Ashamu EA et.al. Significant increase in BW, seminal parameters, testosterone level

and antioxidant activity were observed with SI extract and vitamin C (95).

Thus Sesamum indicum is reported to have analgesic, antioxidant, anti-microbial, anticonvulsant,

anticancer, anti-obesity as well as hepato and nephro protective activities. In addition the

facilitatory effect on fertility in rats has also been reported.

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