the inflammatory process is the response to an injurious stimulus evoked by a wide variety of noxious agents , for example , infections , antibodies or physical injuries .
inflammation is a bodily response to injury , infection or destruction , characterized by heat , redness , pain , swelling and disturbed physiological functions .
inflammation 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 .
the commonly used drug for the management of inflammatory conditions are non - steroidal anti - inflammatory drugs ( nsaids ) , which have several adverse effects especially gastric irritation leading to the formation of gastric ulcers .
of late , traditional medicine is being re - evaluated worldwide , by extensive research on different plant species and their active therapeutic principles .
the rich wealth of the plant kingdom can represent a novel source of newer compounds with significant anti - inflammatory activities .
the major merits of herbal medicine seem to be their perceived efficacy , low incidence of serious adverse effects , 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 .
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 .
non fermented and produced by drying and steaming the fresh leaves ; whereas , black tea leaves are withered , rolled , fermented , and then dried .
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 .
previous researchers have reported the effects of several pharmacological and toxicological properties of green tea and black tea on animals and humans , including anti - inflammatory activities.[712 ] in the present study it has been found worthwhile to evaluate and compare the possible anti - inflammatory effect of green tea and black tea against the denaturation of protein , in vitro , as their anti - inflammatory effects have not been studied in this model so far .
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 fine powder and kept into an air - tight container for use in the study . the powdered plant materials ( 50 g ) were extracted with distilled water ( 350 ml ) by boiling under reflux 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 . the reaction mixture ( 5 ml ) consisted of 0.2 ml of egg albumin ( from fresh hen 's egg ) , 2.8 ml of phosphate - buffered saline ( pbs , ph 6.4 ) and 2 ml of varying concentrations of aqgt and aqbt so that final concentrations became 31.25 , 62.5 , 125 , 250 , 500 , 1000 g / ml . a similar volume of double - distilled water served as the control .
next , the mixtures were incubated at 37  2c in a bod incubator ( labline technologies ) for 15minutes and then heated at 70c for five minutes . after cooling ,
their absorbance was measured at 660 nm ( shimadzu , uv 1800 ) by using the vehicle as a blank .
diclofenac sodium in the final concentrations of ( 78.125 , 156.25 , 312.5 , 625 , 1250 , 2500
g / ml ) was used as the reference drug and treated similarly for the determination of absorbance .
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 .
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 fine powder and kept into an air - tight container for use in the study .
the powdered plant materials ( 50 g ) were extracted with distilled water ( 350 ml ) by boiling under reflux 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 reaction mixture ( 5 ml ) consisted of 0.2 ml of egg albumin ( from fresh hen 's egg ) , 2.8 ml of phosphate - buffered saline ( pbs , ph 6.4 ) and 2 ml of varying concentrations of aqgt and aqbt so that final concentrations became 31.25 , 62.5 , 125 , 250 , 500 , 1000 g / ml . a similar volume of double - distilled water served as the control .
next , the mixtures were incubated at 37  2c in a bod incubator ( labline technologies ) for 15minutes and then heated at 70c for five minutes . after cooling ,
their absorbance was measured at 660 nm ( shimadzu , uv 1800 ) by using the vehicle as a blank .
diclofenac sodium in the final concentrations of ( 78.125 , 156.25 , 312.5 , 625 , 1250 , 2500 g / ml ) was used as the reference drug and treated similarly for the determination of absorbance .
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 )
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 - inflammatory 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 .
therefore , using agents that can prevent protein denaturation would be worthwhile for anti - inflammatory drug development . in the present investigation ,
the in vitro anti - inflammatory effect of aqgt and aqbt was evaluated against denaturation of egg albumin .
the present findings exhibited a concentration - dependent inhibition of protein ( albumin ) denaturation by aqgt and aqbt , throughout the concentration range of 31.25 to 1000 g / ml . diclofenac sodium , in the concentration range of 78.125 to 2500 g / 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 effective than aqbt ; however , the effect of diclofenac sodium was found to be quite low when compared with both the test tea extracts .
effect of aqgt and aqbt against protein denaturation effect of diclofenac sodium against protein denaturation ic50 values of aqgt , aqbt and diclofenac sodium 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 effect by the tea extracts and the reference drug , diclofenac sodium .
black tea and green tea both contain almost a similar amount of flavonoids , however , they differ in their chemical composition ; green tea contains more catechins ( simple flavonoids ) , while the oxidation undergone by the leaves , in order to make black tea , polymerizes these simple flavonoids into theaflavins and thearubigins ( polymerized flavonoids ) . in the present study , the higher anti - inflammatory effect of green tea can be attributed to its higher flavonoids ( catechin ) content .
it has been reported that one of the features of several non - steroidal , anti - inflammatory drugs , is their ability to stabilize ( prevent denaturation ) heat - treated albumin at the physiological ph ( ph : 6.2  6.5 ) .
therefore , from the results of the present preliminary study , it can be concluded that both green and black tea leaves possess a marked anti - inflammatory effect against the denaturation of protein in vitro .
previous researchers have reported the anti - inflammatory activity of both green tea and black tea.[912 ] the present findings 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 different model , in which it has not been studied earlier .
it is suggested that the anti - inflammatory effect of tea leaves could be further evaluated in other experimental models .
 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 . 
INTRODUCTION
MATERIALS AND METHODS
Plant Materials
Preparation of Extracts
Evaluation of Anti-inflammatory Effect 
RESULTS AND DISCUSSION