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Evaluation of anti-inflammatory effects of green tea and black tea: A comparative in vitro study



The present study was conducted to evaluate and compare the anti-inflammatory effects of aqueous extracts of green tea and black tea leaves (Camellia sinensis) against the denaturation of protein in vitro. The test extracts at different concentrations were incubated with egg albumin under controlled experimental conditions and subjected to determination of absorbance to assess the anti-inflammatory property. Diclofenac sodium was used as the reference drug. The present results exhibited a concentration-dependent inhibition of protein (albumin) denaturation by both the tea extracts. From the present findings it can be concluded that both green and black tea leaves possessed a marked anti-inflammatory effect against the denaturation of protein, in vitro. Green tea was found to be more active than black tea, plausibly due to the higher flavonoid contents of green tea.
136 Journal of Advanced Pharmaceutical Technology & Research | Apr-Jun 2012 | Vol 3 | Issue 2
Evaluation of anti-inammatory effects of green tea
and black tea: A comparative in vitro study
The present study was conducted to evaluate and compare the anti-inflammatory
effects of aqueous extracts of green tea and black tea leaves (Camellia sinensis) against
the denaturation of protein in vitro. The test extracts at different concentrations were
incubated with egg albumin under controlled experimental conditions and subjected
to determination of absorbance to assess the anti-inflammatory property. Diclofenac
sodium was used as the reference drug. The present results exhibited a concentration-
dependent inhibition of protein (albumin) denaturation by both the tea extracts. From
the present findings it can be concluded that both green and black tea leaves possessed
a marked anti-inflammatory effect against the denaturation of protein, in vitro. Green
tea was found to be more active than black tea, plausibly due to the higher flavonoid
contents of green tea.
Key words: Anti-inflammatory, green tea, polyphenols, protein denaturation
Priyanka Chatterjee,
Sangita Chandra, Protapaditya Dey,
Sanjib Bhattacharya
Pharmacognosy Division, Bengal School
of Technology (A College of Pharmacy),
Sugandha, Hooghly, West Bengal, India
J. Adv. Pharm. Tech. Res.
short communication
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The inammatory process is the response to an injurious
stimulus evoked by a wide variety of noxious agents,
for example, infections, antibodies or physical injuries.
Inammation is a bodily response to injury, infection or
destruction, characterized by heat, redness, pain, swelling
and disturbed physiological functions. Inammation is
a normal protective response to tissue injury caused by
physical trauma, noxious chemical or microbial agents. It
is the body response to inactivate or destroy the invading
organisms, to remove the irritants, and set the stage for tissue
repair. It is triggered by the release of chemical mediators
from injured tissue and migrating cells.[1] The commonly
used drug for the management of inammatory conditions
are non-steroidal anti-inammatory drugs (NSAIDs), which
have several adverse eects especially gastric irritation
leading to the formation of gastric ulcers.[2,3] Natural products
have contributed signicantly towards the development of
modern medicine. Of late, traditional medicine is being
re-evaluated worldwide, by extensive research on dierent
plant species and their active therapeutic principles. The rich
wealth of the plant kingdom can represent a novel source
of newer compounds with signicant anti-inammatory
activities. The major merits of herbal medicine seem to be
their perceived ecacy, low incidence of serious adverse
eects, and low cost.
Tea, a product made from the leaf and bud of the plant,
Camellia sinensis, is the second most consumed beverage
in the world.[4] Camellia sinensis is a large evergreen
shrub indigenous to Eastern Asia, where it is cultivated
extensively. The dried, cured leaves of C. sinensis have
been used to prepare beverages for more than 4000 years.
The method of curing determines the nature of the tea to
be used for infusion. Green tea is a type of cured tea that is
‘non fermented’ and produced by drying and steaming the
fresh leaves; whereas, black tea leaves are withered, rolled,
fermented, and then dried.[5] Tea has been used medicinally
for centuries in the Traditional Chinese Medicine (TCM). In
recent times, there has been renewed interest in green tea, for
the prevention of several disease risks and other important
health benefits.[6] Previous researchers have reported
the eects of several pharmacological and toxicological
properties of green tea and black tea on animals and
Address for correspondence:
Mr. Sanjib Bhattacharya,
Pharmacognosy Division, Bengal School of Technology
(A College of Pharmacy), Sugandha, Hooghly 712102,
West Bengal, India.
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Chatterjee, et al.: Anti-inammatory effects of green and black tea
Journal of Advanced Pharmaceutical Technology & Research | Apr-Jun 2012 | Vol 3 | Issue 2
humans, including anti-inammatory activities.[7-12] In the
present study it has been found worthwhile to evaluate and
compare the possible anti-inammatory eect of green tea
and black tea against the denaturation of protein, in vitro,
as their anti-inammatory eects have not been studied in
this model so far.
Plant Materials
Packaged green tea and black tea leaves were procured in
the month of July, 2011, from Desai and Sons, Ezra Street,
Kolkata 700001, India. Just after procurement, both type of
tea leaves were ground mechanically into ne powder and
kept into an air-tight container for use in the study.
Preparation of Extracts
The powdered plant materials (50g) were extracted
with distilled water (350 mL) by boiling under reux for
30minutes. The extracts were filtered and evaporated
to dryness to yield the dry extracts of green tea (AQGT,
yield: 51.28%) and black tea (AQBT, yield: 17.45%). The dry
extracts were kept in a vacuum desiccator until use.
Evaluation of Anti-inammatory Eect in vitro
The reaction mixture (5 mL) consisted of 0.2 mL of egg
albumin (from fresh hen’s egg), 2.8 mL of phosphate-buered
saline (PBS, pH 6.4) and 2 mL of varying concentrations of
AQGT and AQBT so that nal concentrations became 31.25,
62.5, 125, 250, 500, 1000 μg/mL. A similar volume of double-
distilled water served as the control. Next, the mixtures
were incubated at 37 ± 2ºC in a BOD incubator (Labline
Technologies) for 15minutes and then heated at 70ºC for
ve minutes. After cooling, their absorbance was measured
at 660 nm (SHIMADZU, UV 1800) by using the vehicle as
a blank. Diclofenac sodium in the nal concentrations of
(78.125, 156.25, 312.5, 625, 1250, 2500 μg/mL) was used as the
reference drug and treated similarly for the determination
of absorbance.[13] The percentage inhibition of protein
denaturation was calculated by using the following formula:
% inhibition = 100 × [Vt / VC - 1]
Where, Vt = absorbance of the test sample, Vc = absorbance
of control.
The extract concentration for 50% inhibition (IC50) was
determined by the dose-response curve.
There are certain problems associated with the use of
animals in experimental pharmacological research, such as,
ethical issues and the lack of rationale for their use when
other suitable methods are available, or can be investigated.
Hence, in the present study the protein denaturation
bioassay was selected for in vitro assessment of the anti-
inammatory property of the aqueous extracts of green
tea and black tea (AQGT and AQBT). Denaturation of
the tissue proteins is one of the well-documented causes
of inflammatory and arthritic diseases. Production of
auto-antigens in certain arthritic diseases may be due to
denaturation of proteins in vivo.[14,15] Therefore, using agents
that can prevent protein denaturation would be worthwhile
for anti-inammatory drug development.
In the present investigation, the in vitro anti-inammatory
eect of AQGT and AQBT was evaluated against denaturation
of egg albumin. The results are summarized in Table 1.
The present ndings exhibited a concentration-dependent
inhibition of protein (albumin) denaturation by AQGT and
AQBT, throughout the concentration range of 31.25 to 1000
μg/mL. Diclofenac sodium, in the concentration range of
78.125 to 2500 μg/mL, was used as a reference drug which
also exhibited concentration-dependent inhibition of protein
denaturation [Table 2]. The IC50 values are summarized in
Table 3. Here, AQGT was found to be more eective than
AQBT; however, the eect of diclofenac sodium was found to
be quite low when compared with both the test tea extracts.
This was further conrmed by comparing their IC50 values
[Table 3].
The increments in the absorbance of the test samples, with
respect to the control, indicated stabilization of protein,
that is, inhibition of protein (albumin) denaturation or an
anti-denaturation eect by the tea extracts and the reference
drug, diclofenac sodium.[16]
Tea leaves contain varying amounts of polyphenols
Table 1: Effect of AQGT and AQBT against
protein denaturation.
% Inhibition
% Inhibition
Control - -
31.25 220 140
62.5 320 180
125 620 220
250 1100 700
500 2480 1540
1000 4980 2480
AQGT: Aqueous extract of green tea, AQBT: Aqueous extract of black tea
Table 2: Effect of diclofenac sodium against
protein denaturation
Concentration (µg/mL) % Inhibition
Control -
78.125 12.5
156.25 12.5
312.5 25
625 50
1250 212.5
2500 812.5
AQGT: Aqueous extract of green tea, AQBT: Aqueous extract of black tea
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Chatterjee, et al.: Anti-inammatory effects of green and black tea
138 Journal of Advanced Pharmaceutical Technology & Research | Apr-Jun 2012 | Vol 3 | Issue 2
particularly flavonoids. Polyphenols are well-known
natural products known to possess several notable
biological properties.[17] Black tea and green tea both contain
almost a similar amount of flavonoids, however, they
dier in their chemical composition; green tea contains
more catechins (simple avonoids), while the oxidation
undergone by the leaves, in order to make black tea,
polymerizes these simple avonoids into theaavins and
thearubigins (polymerized avonoids).[8,18] In the present
study, the higher anti-inammatory eect of green tea can
be aributed to its higher avonoids (catechin) content.
The eect may be due to the synergistic eect rather than
a single constituent.
It has been reported that one of the features of several
non-steroidal, anti-inammatory drugs, is their ability to
stabilize (prevent denaturation) heat-treated albumin at
the physiological pH (pH: 6.2 – 6.5).[19] Therefore, from
the results of the present preliminary study, it can be
concluded that both green and black tea leaves possess a
marked anti-inammatory eect against the denaturation of
protein in vitro. Previous researchers have reported the anti-
inammatory activity of both green tea and black tea.[9-12] The
present ndings corroborated this property in vitro, against
this protein denaturation model. This correlation is further
strengthened by the fact that the present study was performed
in a dierent model, in which it has not been studied earlier.
It is suggested that the anti-inammatory eect of tea leaves
could be further evaluated in other experimental models.
The authors are thankful to the authority of the Bengal School of
Technology (A College of Pharmacy), Sugandha, Hooghly, 712102,
West Bengal, India, for providing the necessary facilities for the
present study.
1. Anonymous. New medical dictionary. 2nd ed. New Delhi: Oxford
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9. Aneja R, Odoms K, Denenberg AG, Wong HR. Theaavin, a black
tea extract, is a novel anti-inammatory compound. Crit Care Med
10. Nag Chaudhuri AK, Karmakar S, Roy D, Pal S, Pal M, Sen T.
Anti-inammatory activity of Indian black tea (Sikkim variety).
Pharmacol Res 2005;51:169-75.
11. Tipoe GL, Leung TM, Hung MW, Fung ML. Green tea
polyphenols as an anti-oxidant and anti-inammatory agent
for cardiovascular protection. Cardiovasc Hematol Disord Drug
Targets 2007;7:135-44.
12. Cavet ME, Harrington KL, Vollmer TR, Ward KW, Zhang JZ. Anti-
inammatory and anti-oxidative eects of the green tea polyphenol
epigallocatechin gallate in human corneal epithelial cells. Mol Vis
13. Chandra S, Chaerjee P, Dey P, Bhaacharya S. Evaluation of anti-
inammatory eect of ashwagandha: a preliminary study in vitro.
Phcog J 2012;4:47-9.
14. Opie EL. On the relation of necrosis and inflammation to
denaturation of proteins. J Exp Med 1962;115:597-608.
15. Umapathy E, Ndebia EJ, Meeme A, Adam B, Menziwa P, Nkeh-
Chungag BN, et al. An experimental evaluation of Albuca setosa
aqueous extract on membrane stabilization, protein denaturation
and white blood cell migration during acute inammation. J Med
Plants Res 2010;4:789-95.
16. Jagtap VA, Agasimundim YS, Jayachandran E, Sathe BS.
In vitro anti-inflammatory activity of 2-amino-3-(substituted
benzylidinecarbohydrazide)-4,5,6,7 tetrahydrobenzothiophenes.
J Pharm Res 2011;4:378-79.
17. Bhaacharya S. Are we in the polyphenols era? Pharmacognosy
Res 2011;3:147.
18. Graham H. Green tea consumption, consumption and polyphenol
chemistry. Prev Med 1992;21:334-50.
19. Williams LAD, O’Connar A, Latore L, Dennis O, Ringer S,
Whiaker JA, et al. The in vitro anti-denaturation eects induced
by natural products and non-steroidal compounds in heat treated
(immunogenic) bovine serum albumin is proposed as a screening
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West Indian Med J 2008;57:327-31.
Table 3: IC50 values of AQGT, AQBT and
diclofenac sodium
Treatments IC50 (µg/mL)
AQGT 7.10
AQBT 11.16
Diclofenac sodium 625
AQGT: Aqueous extract of green tea, AQBT: Aqueous extract of black tea
How to cite this article: Chatterjee P, Chandra S, Dey P,
Bhaacharya S. Evaluation of anti-inammatory eects of green
tea and black tea: A comparative in vitro study. J Adv Pharm Tech
Res 2012;3:136-8.
Source of Support: Nil, Conict of Interest: Nil.
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... Several anti-inflammatory drugs have shown dose-dependent ability to inhibit thermally-induced protein denaturation [33]. The commonly used drugs for the management of inflammatory conditions are non-steroidal anti-inflammatory drugs, which have several adverse effects, especially gastric irritation leading to the formation of gastric ulcers [34]. Therefore, the search for natural antioxidants with anti-inflammatory activity has greatly increased in recent years. ...
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The Ipomoea quamoclit is known as konjolata, commonly called the cypress vine Convolvulaceae family. I. quamoclit leaves found several medicinal uses such as antioxidant, anticancer, anti-inflammatory, etc. It is also traditionally used as an antidote to snake bites. Aerial parts of the plant are selected to evaluate the comparative phytochemical screening, antioxidants such as DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay, total phenolic content, total flavonoids content, total antioxidant capacity, cytotoxicity, and in-vitro anti-inflammatory activity of the nonpolar and polar solvent extracts. The aerial part of the plant was successively extracted with dichloromethane (DCM), ethyl acetate, methanol, and 95% ethanol. Phytochemical screening of the extracts of I. quamoclit reveals the presence of components such as alkaloids, flavonoids, saponins, tannins, steroids, carbohydrates, etc. The results of the antioxidant study exhibit good antioxidant properties in different methods such as DPPH free radical scavenging, total phenolic content, total flavonoids content, and the total antioxidant capacity of the extracts of I. quamoclit. In which nonpolar solvent DCM extract showed significant antioxidant activity compared to the other extracts. Cytotoxicity study of the extracts of I. quamoclit indicates that the ethyl acetate and methanolic extract have good cytotoxic activity compared to other extracts LC 50 6.092 and 1.688 µg/mL, All the extracts of the polar and nonpolar solvent of I. quamoclit inhibit thermal denaturation of protein is possibly a contributing factor for its anti-inflammatory activity. The extracts (25-200μg/ml) showed significant inhibition of denaturation of egg albumin and bovine albumin in a concentration-dependent manner, highest inhibition found at 200μg/mL at 98.07%. The finding of the study suggested that nonpolar solvents dichloromethane has good antioxidant property, and the polar solvents methanol extract has good cytotoxic and anti-inflammatory activity.
... Regardless of the tea type, several lines of evidence suggest that tea polyphenols are bioactive molecules. The bioactivity of tea is not merely limited to basic antioxidant and metal chelating activities, but rather tea polyphenols have been shown to confer strong immunostimulatory [28], anti-inflammatory [29], anti-cancerous [30], antibacterial [31], and antiviral effects [32]. In fact, tea phenols have also been implicated as potential protective agents against the ongoing pandemic of SARS-CoV-2 virus [33][34][35]. ...
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A cellular senescence-centric understanding of biological aging and age-related chronic diseases is rapidly emerging. As a result, strategies aimed at mitigating the deleterious aspects of cellular senescence are increasingly becoming desirable. Tea is a globally admired and nutrient-rich beverage that not only refreshes the senses but is also implicated in several health beneficial effects including the extension of organismal healthspan and lifespan. The present review discusses the emerging anti-cellular senescence attributes of tea consumption and provides a perspective that the anti-aging aspects of tea should be studied in the purview of cellular senescence. Current understanding of the integrative effects of the immune system and gut microbiome on cellular senescence have also been discussed with the rationale of mitigatory effects of tea. Future research directions and recommendations have been provided which may ultimately help augment tea-oriented successful and healthy aging approaches.
... [5][6][7] These alkaloids and flavonoids are rich with antioxidant properties which could reverse the STZ-induced oxidative impact in diabetic rats. [17] To the best of the authors' comprehension, this is the first scientific account of the antidiabetic potential of this plant. Form the present experiments it may be inferred that, the stem bark of Premna spinosa possesses significant effectiveness against metabolic disorders like diabetes mellitus hence, further exploration on this plant leading to isolation and molecular mechanism studies are required to reap its exact usefulness for the therapeutic management of human diabetes mellitus. ...
The purpose of the study is to evaluate the antidiabetic and hyperlipidemic potential of stem bark extract of Premna spinosa (Lamiaceae), by using streptozotocin (STZ)-nicotinamide (NA)-induced diabetic and triton-induced hyperlipidemic models in albino rats. The blood glucose, total cholesterol, and triglyceride levels were determined in STZ-NA-induced diabetic and triton-induced hyperlipidemic rats, as per the respective protocols. It was found that there is the dose dependent and significant reduction in foregoing parameters on the administration of extract from Premna spinosa stem bark at the doses of 200, 400, and 800 mg/kg body weight to diabetic and hyperlipidemic rats. From these observed results it may be inferred that the stem bark of Premna spinosa possesses remarkable antidiabetic and antihyperlipidemic properties.
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Albuca setosa is widely distributed in the Eastern Cape region of South Africa where its traditional usage is very extensive. This study was aimed to experimentally evaluate the effect of A. setosa water extract (ASWE) on inflammation events such as membrane stabilization, protein denaturation and white blood cell migration during acute inflammation. This study was undertaken using hypotonicity and heat induced erythrocytes haemolysis, heat induced albumin denaturation, carrageenan and dextran induced peritonis and rat paw edema. The results showed that ASWE at a concentration range of 125 -500 g/ml significantly (p < 0.01) protects the erythrocyte membrane against lysis induced by heat and hypotonic medium solution. At the dose of 500 and 125 g/ml, ASWE showed significant (p < 0.01) inhibition of 59 and 65% of protein denaturation of egg albumin. Oral administration of 150 and 300 mg/kg of ASWE significantly (p < 0.05) reduced the total WBC count in rat paw fluid after inflammation induced by carrageenin and in the peritoneal wash after acute inflammation induced by dextran and carrageenan, respectively. The present work contributes to the validation of the anti-inflammatory activity of the plant and may provide some evidence for its folk use and further exploitation.
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To determine the anti-inflammatory and anti-oxidant effects of epigallocatechin gallate (EGCG), the major polyphenol component of green tea, in human corneal epithelial cells (HCEpiC). HCEpiC were challenged with interleukin-1β (IL-1β) for 18 h or hyperosmolarity (440 mOsm) for 24 h. Luminex technology was used to determine the effects of EGCG (0.3-30 µM) on IL-1β- or hyperosmolar-induced cytokine release into the medium. Cell metabolic activity was measured using the alamarBlue assay. Effects of EGCG on mitogen-activated protein kinase (MAPK) phosphorylation were determined by cell-based enzyme-linked immunosorbent assay (ELISA) and western blotting. Effects of EGCG on nuclear factor kappa B (NFκB) and activator protein-1 (AP-1) transcriptional activity were assessed by reporter gene assay. The effects of EGCG on glucose oxidase (GO)-induced reactive oxygen species (ROS) production was determined using the ROS probe CM-H₂DCFDA. Treatment of HCEpiC with 1 ng/ml IL-1β for 18 h significantly increased release of the cytokines/chemokines granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1), while hyperosmolarity-induced release of IL-6 and MCP-1. When cells were treated with IL-1β and EGCG or hyperosmolarity and EGCG there was a dose-dependent reduction in release of these cytokines/chemokines, with significant inhibition observed at 3-30 µM. There was no effect of EGCG on cell metabolic activity at any of the doses tested (0.3-30 µM). EGCG significantly inhibited phosphorylation of the MAPKs p38 and c-Jun N-terminal kinase (JNK), and NFκB and AP-1 transcriptional activities. There was a significant dose-dependent decrease in GO-induced ROS levels after treatment of HCEpiC with EGCG. EGCG acts as an anti-inflammatory and anti-oxidant agent in HCEpiC and therefore may have therapeutic potential for ocular inflammatory conditions such as dry eye.
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There are emerging ethical issues with regards to the use of animals in the early stages of drug discovery for anti-inflammatory and degenerative diseases from natural products using the activity-directed isolation pathways when many compounds (eg > 100) are present in the crude extract or fraction and are to be tested The above-mentioned is the main reason for proposing the use of the in vitro anti-denaturation (stabilization) effects of heat treated (immunogenic) bovine serum albumin (BSA) as an assay. Current methods used for detecting and isolating a wide range of anti-inflammatory compounds in the early stages of the drug discovery process utilize a large number of animals. When BSA is heated and is undergoing denaturation, it expresses antigens associated to Type III hypersensitive reaction and which are related to diseases such as serum sickness, glomerulonephritis, rheumatoid arthritis and systemic lupus erythematosus. Thus, the assay that is being proposed should be applicable to the discovery of drugs for treating the above mentioned diseases and others, once the compounds stabilize the denaturation process.
Known for their ease of use, artful presentation of scientific information, and evidence-based approach, James Duke's comprehensive handbooks are the cornerstone in the library of almost every alternative and complementary medicine practitioner and ethnobotanist. Using the successful format of these bestselling handbooks, Duke's Handbook of Medicinal Plants of the Bible covers 150 herbs that scholars speculate, based on citations, were used in Biblical times.
Tea is an important dietary source of flavanols and flavonols. In vitro and animal studies provide strong evidence that tea polyphenols may possess the bioactivity to affect the pathogenesis of several chronic diseases, especially cardiovascular disease and cancer. However, the results from epidemiological and clinical studies of the relationship between tea and health are mixed. International correlations do not support this relationship although several, better controlled case-referent and cohort studies suggest an association with a moderate reduction in the risk of chronic disease. Conflicting results between human studies may arise, in part, from confounding by socioeconomic and lifestyle factors as well as by inadequate methodology to define tea preparation and intake. Clinical trials employing putative intermediary indicators of disease, particularly biomarkers of oxidative stress status, suggest tea polyphenols could play a role in the pathogenesis of cancer and heart disease.
Introduction: Ashwagandha (Withania somnifera) is an important medicinal plant in Indian traditional system of medicine and traditionally has been used for several medicinal purposes in India. The present study was conducted to evaluate the anti-inflammatory effect of hydroalcoholic extract of ashwagandha against denaturation of protein in vitro. Methods: The test extract at different concentrations was incubated with egg albumin in controlled experimental conditions and subjected to determination of absorbance and viscosity to assess the anti-inflammatory property. Diclofenac sodium was used as the reference drug. Results: The present results exhibited a concentration dependent inhibition of protein (albumin) denaturation by the ashwagandha extract. The effect of diclofenac sodium was found to be less when compared with the test extract. Conclusion: Form the present findings it can be concluded that ashwagandha possessed marked anti-inflammatory effect against denaturation of protein in vitro. The effect was plausibly due to the alkaloid and withanolide contents of ashwagandha.