Performing your original search, THEAFLAVIN PROTECT BRAIN CELL, in PubMed Central will retrieve 14 records. Mediators Inflamm.

2006; 2006 !"# $0%&0. PMCI*# PMC(6!+0++ Published online 2006 'ctober &. doi# (0.((!!)MI)2006)$0%&0 Co,yright - 2006 .ei Cai et al.

Theaflavin Ameliorates Cerebral Ischemia-Reperfusion Injury in Rats Through Its Anti-Inflammatory Effect and Modulation of STAT.ei Cai,(, 2 Cai/0ong 1i,2 2i/1iang 3u,24 2ian/5uo Chen,( Chao 1iu,( 6ing Min,2 3ei 7u,2 Chang/8an 'uyang,2 and 2in/8e Chen$
*e,artment of Pharmacology, 9ong:i Medical College, 8ua;hong <niversity of =cience and 9echnology, ($ 8ang>ong 0oad, 3uhan %$00$0, China 2 *e,artment of Pharmacology, Medical College, ?ianning <niversity, $ 5uihua 0oad, ?ianning %$+(00, China $ *e,artment of Pharmacology, Medical College, 3uhan <niversity, $& *onghu 0oad, 3uhan %$00+(, China 42i/1iang 3u# @mail# :iliangAwByahoo.com.cn 0eceived March $0, 2006; 0evised Cugust $0, 2006; Ccce,ted Cugust $0, 2006. 9his is an o,en access article distributed under the Creative Commons Cttribution 1icense, which ,ermits unrestricted use, distribution, and re,roduction in any medium, ,rovided the original wor> is ,ro,erly cited.
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Abstract Theaflavin, a major constituent of black tea, possesses biological functions such as the antioxidative, antiviral, and anti-inflammatory ones. The purpose of this study was to verify whether theaflavin reduces focal cerebral ischemia injury in a rat model of middle cerebral artery occlusion (M !"#. Male $prague-%awley rats were anestheti&ed and subjected to ' hours of M !" followed '( hours reperfusion. Theaflavin administration (), *+, and '+ mg,kg, -.# ameliorated infarct and edema volume. Theaflavin inhibited leukocyte infiltration and expression of - !M-*, "/-', and i0"$ in injured brain. 1hosphorylation of $T!T-*, a protein which mediates intracellular signaling to the nucleus, was enhanced '-fold over that of sham group and was inhibited by theaflavin. "ur study demonstrated that theaflavin significantly protected neurons from cerebral ischemia-reperfusion injury by limiting leukocyte infiltration and expression of - !M-*, and suppressing upregulation of inflammatory-related prooxidative en&ymes (i0"$ and "/-'# in ischemic brain via, at least in part, reducing the phosphorylation of $T!T-*.
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INTRODUCTION !cute ischemic stroke is the leading cause of adult disability and it is also an important cause of death in industriali&ed countries with a high incidence affecting up to +.'2 of the population every year 3*4. !lthough pathologic mechanisms

leading to cerebral ischemic injury remained unclear, it has been emphasi&ed that inflammatory process had fundamental roles in both the etiology of ischemic cerebrovascular disease and the pathophysiology of cerebral ischemia 3', 54. 0eutrophils are critically involved in the early stage of inflammatory reaction after ischemia, initiating scavenger functions which are later subsumed by macrophages 3(, )4. 6ndothelial cells actively participate in inflammatory events by regulating leukocyte recruitment via the expression of inflammation-related genes such as - !M-*, . !M-*, 6-selectin, -7-8, -7-9, and cyclooxygenase-' 38, :4. yclooxygenase ( "/#, a rate-limiting en&yme in the metabolism of arachidonic acid into prostanoids, produces 1;<' which in subse=uent steps gives rise to 1;s with various physiological functions 39>*+4. -t has been demonstrated in previous reports that cerebral ischemia upregulated the inducible form of "/ ( "/-'# in neurons, glial cells and infiltrating leukocytes in injured brain 3**>*54. -nhibition of "/-' activity during or after ischemia and genetic deletion of "/-' reduce infarct volume 3*(4. -n addition, neuronal overexpression of "/-' increases cerebral infarction 3*), *84. These observations suggest that "/-' plays a deleterious role in cerebral ischemia. -nterestingly, nitric oxide produced by inducible form of nitric oxide synthase (i0"$# has been found to positively regulate "/-' activity in focal cerebral ischemia 3*:4. erebral ischemia enhanced i0"$ expression in neurons, endothelial cells, and microglia 3*9, *?4. i0"$ clearly plays a role in stroke outcome, as evidenced by its selective inhibition in the rat model of M !" or its genetic deletion 3'+, '*4. $T!T-* is a member of the signal transducers and activators of transcription proteins family ($T!Ts#, which mediate intracellular signaling initiated at cytokine cell surface receptors and transmit to the nucleus. The terminal domains of $T!T proteins contain a transcriptional transactivation domain which is essential for maximal $T!T function. @ecent study has shown that myocardial ischemia activates A!Bs, followed by recruitment of $T!T-*, resulting in transcriptional upregulation of inducible nitric oxide synthase (i0"$# and cyclooxygenase-' ( "/-'# 3''>'(4. -t has been demonstrated that focal cerebral ischemia induced $T!T-* activation 3')4 and $T!T-* knockout mice developed smaller lesions and less pronounced neurological deficit following transient focal cerebral ischemia 3'84. atechins and theaflavins are two groups of natural polyphenols found in green tea and black tea, respectively 3':4. These tea polyphenols possess a broad spectrum of biological functions such as antioxidative, antibacteria, antitumor, antiviral, antiinflammatory, and cardiovascular protection activities 3'9>5*4. -t has been reported that epigallocatechin-5-gallate (6; ;#, a major constituent of catechins, may attenuate cerebral ischemia-reperfusion injury in rats 35'4 and it was also a potent inhibitor of $T!T-* phosphorylation 3554. Therefore, the present study was

undertaken to evaluate the neuronal protective potential of theaflavin (TC*#, a major constituent of theaflavins, in middle cerebral artery occlusion (M !"# induced focal cerebral ischemia-reperfusion model in rats.
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MATERIALS AND METHODS E"perimental protocol and drug Male $prague-%awley rats weighing ''+>'8+ g were housed at room temperature under a controlled *' h light,dark cycle and allowed access to food and water ad libitum. !ll experiments were performed as approved by the institutional animal care and use committee. @ats were divided into six groups and each group had ten animals. The first was vehicle-treated group, that is, ischemia was induced for ' h of M !" followed by reperfusion for '( h. The second was sham group. The theaflavin (a pure natural product collection provided by Micro$ource %iscovery $ystem -nc, ;aylordsville, onn#-treated groups were separated into a low dosage group (TC*, ) mg,kg#, a middle dosage group (TC*, *+ mg,kg#, and a high dosage group (TC*, '+ mg,kg#. The intravenous injection of theaflavin was conducted directly before the reperfusion. The sixth was nimodipine-treated group (* mg,kg, -1#. !ll other chemicals and reagents were of the highest analytical grades available locally. Middle cerebral artery occlusion induced focal cerebral ischemia Cocal cerebral ischemia was produced by occluding the left middle cerebral artery according to the methods by 7onga et al 35(4. Driefly, the rats were anaestheti&ed with hloral hydrate ((++ mg,kg, -1#. Through a midline neck incision, the left common and external carotid artery were isolated from muscles and coagulated. ! 5-+ nylon suture with a blunted tip was inserted into the internal carotid through the external carotid artery stump and advanced up to '* mm or till resistance was left. !fter ' h of M !", the suture was removed to restore blood flow. -n sham group, the same surgical procedure was performed except that the suture was introduced into the external carotid artery but not advanced. !fter surgery, the incision was sutured and the rats were returned to their cage with free access to water and food. Twenty-four hours after reperfusion, rats were sacrificed by rapid decapitation under deep anesthesia and the brains were taken out for biochemical estimations. Infarct and edema volume Twenty-four hours after reperfusion, whole brains were rapidly removed. -mmediately after being weighed, the brains were sliced into '-mm-thick coronal sections and stained with '2',5,)-triphenyltetra&oliumchloride (TT , $igma!ldrich# at 5:E for 5+ minutes in the dark, followed by fixation with *+2 formalin

at room temperature overnight. The sections were photographed with a digital camera (Bodak % '(+, F$!# connected to a computer. The unstained areas, defined as infarct tissue, were calculated by using an image analysis program (!dobe 1hotoshop ).+ $#. The infarct volume was calculated by measuring the unstained area in each slice. 6dema correction of infarct volume was done using the e=uation, volume correction G (infarct volume H contralateral volume#,ipsilateral volume. The volumes of both the hemispheres were calculated from which edema volume was calculated by subtracting the contralateral volume from the ipsilateral volume. Measurement of lipid pero"idation The estimate of lipid peroxidation of the cerebral cortex was determined by measuring the formed malondialdehyde (M%!#. Driefly, brain tissues were homogeni&ed (*+2, w,v# with cold *.)2 B l. The homogenate was mixed with a *2 phosphoric acid and 82 TD! ($igma-!ldrich# a=ueous solution. The mixture was heated for () minutes in a boiling water bath. !fter cooling, n-butanol was added and mixed vigorously. The absorbance of the butanol phase was measured at )') nm. ! serially diluted M%! ($igma-!ldrich# solution was prepared and used as a standard. The data (M%!# was expressed as nmol,mg protein. Myelopero"idase assay The activity of myeloperoxidase (M1"# was determined as an indicator of 1M0s migration, as previously described 35)4. The method of assaying M1" activity was according to the guide of the assay kit (0anjing Aiancheng Dioengineering o 7td, hina#. Immunohistochemistry detection The procedures were processed according to the protocols recommended for - !M*, i0"$, and "/-' immunohistochemistry kit. Collowing deparaffini&ation and rehydration, the cortices sections were exposed to 52 hydrogen peroxide for *+ minutes to bleach endogenous peroxidases. Then microwave oven-based antigen retrieval was performed. $lides were probed with either anti-- !M-* (* I *++, rat monoclonal, $anta ru& Diotechnology#, anti-i0"$ (* I *++, rat polyclonal, $anta ru& Diotechnology#, or anti- "/-' (* I )+, rat monoclonal, $anta ru& Diotechnology# for * hour at 5:E , washed 5 times in 1D$, incubated with biotinlabeled anti-rat -g; for * hour at 5:E , respectively. -ncubation with 1D$ instead of the primary antibody served as a negative control. !fter washing in 1D$, tissues were visuali&ed with 5, 5J-diaminoben&idine tetrahydrochloride (%!D# and counterstained with hematoxylin. Cinally, the sections were dehydrated in graded ethanol, immersed in xylene and coverslipped. -n specimens the positive cells were counted in cortex in ten randomly selected areas from each case and expressed as

number of immunopositive,mm'. @esults are presented as mean K $6M.

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RT-PCR Total @0! was extracted from cortex using T@-&ol reagent ($igma o#. c%0! was synthesi&ed according to the manufacturerLs instruction of reverse transcription kit (;-D "-D@7, F$!#, and then amplified with a multiplex 1 @ kit (;-D "-D@7, F$!#. onditions for amplification were as followsI initial denaturation for ' minutes at ?(E , 5) cycles of ?(E for 5+ seconds, 8+E for () seconds, :'E for 8+ seconds, and a final extension stop at :'E for : minutes. The rat specific primers (sense and antisense primers# for i0"$, "/-', and M-actin were )J;;T; T;T!TTT TT! ;!;; ;!!;!!;;-5J and )J;;T; T;T T;TT!;;!;;T !!;T!!!;;; -5J (i0"$,')? bp#N )J!T;T !!!!- ;T;;T;!!T;-5J and )J-!T;;;!;TT;;; !;T !T !;-5J ( "/-',5:( bp#N )!T;;!T;! ;!T!T ; T;-5 and )-!T;!;;T!;T T;T !;;T-5 (M-actin, )89 bp#, respectively. @eaction products were then separated on a *.)2 agarose gel, stained with ethidium-bromide, and visuali&ed by F. transillumination. <1-!$*+++ software analysis system was used to determine the relative absorbance of m@0! expression. #estern blot analysis The cortices of brains were removed and used for Oestern analysis. 1rotein concentrations were determined using the Dio-@ad protein assay kit (Dio-@ad, <ercules, alif# and all samples were adjusted to an e=ual protein content before analysis. $amples (5+ Pg of total protein# were separated on 92 denaturing polyacrylamide gel. Collowing electrophoresis, proteins were transferred to a nitrocellulose membrane (9+ ., ?+ minutesN transfer buffer ') mM Tris, *?+ mM glycine, '+2 methanol, +.)2 sodium dodecyl sulfate# by an electroblotter (Dio-rad#. !fter being blocked for two hours at room temperature in blocking buffer ()2 nonfat milk in '+ mM Tris,< l, p< :.8, *(+ mM 0a l, +.)2 Tween '+#, membranes were incubated over night at (E with primary antibodies against antiphospho-$T!T-*Tyr-:+* (Qymed, $outh $an Crancisco, alif#, or anti-$T!T-* ($anta ru& Diotechnology, $anta ru&, alif#. Membranes were then washed (in '+ mM Tris,< l, p< :.8, *(+ mM 0a l, +.*2 Tween '+# and incubated with a peroxidase-conjugated secondary antibody at room temperature for )+ minutes. The immunoblots were visuali&ed using Oestern blotting luminal reagent ( ell $ignal orp#. The density of protein band was scanned and analy&ed with an image analy&er.

Statistical analysis Fnless otherwise stated, all the results were finally presented as means K $6M. $tatistical differences between different groups were assessed by a one-way analysis of variance and $tudent-0ewman-Beuls test. P value less than .+) was considered statistically significant.
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RESULTS Effect of theaflavin on cerebral infarction and edema -nfarct volume was measured in the coronal brain sections which were stained with TT . Two hours of M !" and '( hours of reperfusion showed an infarct volume of ''+.9: K ':.(' mm5. The infarct volume was decreased to *95.(? K *?.55 mm5, *5?.+8 K **.'9 mm5, and **9.') K *+.58 mm5 in ), *+, and '+ mg,kg theaflavintreated rats, respectively, (Cigure *#. Theaflavin at the doses of *+ and '+ mg,kg produced (+.:? K 9.:*2 and )'.5+ K ?.:?2 reduction in infarct volume, respectively, as compared to vehicle-treated group (P R .+*, Cigure *#. Two hours of M !" and '( hours of reperfusion resulted in *55.85 K **.+: mm5 increase in the ipsilateral volume due to edema. Theaflavin at the doses of ), *+, and '+ mg,kg resulted in reduction of edema volume to ?9.8* K ').5( mm5, 8*.5: K *(.*5 mm5, and )*.') K ?.?: mm5 of ipsilateral hemisphere, respectively (Cigure *#. Theaflavin at the doses of *+ and '+ mg,kg showed (9.85K:.9(2 and )).+(K9.+*2 reduction in edema volume, respectively as compared to vehicle-treated group (P R .+*, Cigure *#. 0imodipine-treatment also reduced the infarct and edema volumes. The infarct and edema volumes of 0imodipine-treated group were **+.5? K *+.*: mm5 and )).'8 K *+.8) mm5, respectively (P R .+*, Cigure *#.
Figure 1 (a# @epresentative coronal brain sections stained with TT after ' hours of M !" and '( hours of reperfusion showing infarction. %ark-colored region in the TT stained sections indicated nonischemic portion of brain and pale-colored region indicated ischemic (more ...#

Effect of theaflavin on M$A The level of M%! significantly increased in the vehicle-treated group more than in the sham group. !s compared to the vehicle-treated group, the levels of M%! significantly decreased in the theaflavin and nimodipine-treated groups (P R . +*, Table *#. The theaflavin-treated group ('+ mg,kg# had the same effect as compared to the nimodipine-treated group (P S .+)#. <owever, the M%! levels of theaflavin-treated groups were still higher than that of sham group.

Table 1 6ffect of theaflavin on M%! and M1" activities (

K s nG *+#.

Effect of theaflavin on inflammatory injury of cerebral ischemia -nfiltration of leukocytes to -,@-injured tissue provides predominant sources for M1", an important prooxidative en&yme responsible for oxidative stress in -,@injured brain. -n this study, the M1" activity was relatively low in the sham group, and significantly increased in the vehicle-treated group. Treatment with *+ and '+ mg,kg theaflavin significantly reduced M1" activity in the -,@-injured cerebral tissue. 0imodipine-treatment also reduced M1" activity (Table *#. Effect of theaflavin on ICAM- % i&'S% and C'(-) protein production The protein expressions of - !M-*, i0"$, and "/-' in the ischemic cortex of the vehicle-treated group significantly increased compared with those of the sham group. The expression of - !M-* was obviously identified on the microvascular endothelial cells in the ischemic hemisphere (Cigure '#. The positive cells of i0"$ and "/-' were found with brown cytoplasma and predominantly located within the neurons, glial cells, and infiltrating leukocytes (Cigures (Cigures5,5, ,(#.(#. The protein expressions of - !M-*, i0"$, and "/-' decreased dose dependently in theaflavin-treated groups (Table '#. 6ffect of '+ mg,kg theaflavin was similar to that of nimodipine (* mg,kg#.
Figure 2 -mmunohistochemical staining of - !M-* in brain tissues of (a# vehicle-treated rats and (b# theaflavin-treated rats ('+ mg,kg#, $1H(++. - !M-* protein is mainly expressed on the microvascular endothelial cells. - !M-* expression decreases dramatically (more ...# Figure 3 -mmunohistochemical staining for i0"$ protein expression in (a# vehicle-treated rats and (b# theaflavin-treated rats ('+ mg,kg#, $1 H (++. The number of i0"$ immunoreactive positive cells in theaflavin-treated groups is significantly less than (more ...# Figure 4 -mmunohistochemical staining for "/-' protein expression in (a# vehicle-treated rats and (b# theaflavin-treated rats ('+ mg,kg#, $1 H (++. The number of "/-' immunoreactive positive cells in theaflavin group is significantly less than that of (more ...#

Table 2 - !M-*, i0"$, and "/-' protein production in vehicle and theaflavin-treated groups ( K s n G *+#.

Effect of theaflavin on C'(-) and i&'S mR&A e"pressions The m@0! expressions of "/-' and i0"$ were analy&ed by @T-1 @. The brain tissue obtained from the sham group showed low m@0! expression levels of "/' and i0"$. !fter ' hours of M !" and '( h reperfusion, the expressions of "/' and i0"$ remarkably increased in ischemic hemisphere in the vehicle-treated group as compared with the sham group. Theaflavin-treatment could reduce molecule m@0! expressions dose dependently and nimodipine also reduced the expressions of molecule m@0! (Cigures (Cigures)), ,88#.
Figure 5 (a# The m@0! expression of "/-' was assessed by using @T-1 @ as standardi&ed by coamplifying the housekeeping gene M-actin. 7anes *>:I Marker, .ehicle, $ham, TC* () mg T kgU*#, TC* (*+ mg T kgU*#, TC* ('+(more ...# Figure 6 (a# The m@0! expression of i0"$ was assessed by using @T-1 @ as standardi&ed by coamplifying the housekeeping gene M-actin. 7anes *>:I Marker, .ehicle, $ham, TC* () mg T kgU*#, TC* (*+ mg T kgU*#, TC* ('+(more ...#

Effect of theaflavin on STAT- protein e"pression The levels of $T!T-* phosphorylation on tyrosine :+* were markedly enhanced in brains subjected to ' hours of M !" followed '( hours reperfusion. <owever, the brains treated with theaflavin and nimodipine reduced $T!T-* phosphorylation levels on tyrosine :+* (Cigure :#. Theaflavin-treatment could reduce $T!T-* phosphorylation dose dependently. These results demonstrate that theaflavin could have the ability to inhibit $T!T-* :+* phosphorylation as well as protect brain against -,@-induced inflammation.
Figure 7 (a# Oestern blotting showed levels of $T!T-* in brain tissue of rats. 7anes *>8I .ehicle, $ham, TC* () mg T kgU*#, TC* (*+ mg T kgU*#, TC* ('+ mg T kgU*#, 0imodipine. (b# $tatistical analysis revealed (more ...#
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DISCUSSION -n the current study theaflavin-treatment showed protective effects on brain injuries induced by middle cerebral artery occlusion followed by reperfusion in rats by blocking inflammation-related events (M1" and - !M-*# and expressions of prooxidative en&ymes such as "/-' and i0"$. Curther, the protective effect of

theaflavin was associated with downregulation of $T!T-* phosphorylation. The neuronal protective potential of theaflavin was dose dependently and the effect of '+ mg,kg theaflavin was similar to that of nimodipine. @ats subjected to cerebral ischemia-reperfusion showed typical markers of cerebral inflammation and oxidative,nitrosative injury including leukocyte infiltration into the infarct area (enhanced M1" activity#, upregulation of adhesion molecules (- !M-*#, and induction of prooxidative en&ymes ( "/-' and i0"$# 358, 5:4. -schemia activates a cascade that leads to the induction and expression of genes in a variety of cell types throughout the central nervous system ( 0$#. "/-', one product of such immediate early genes, has become the focus of attention because it is the rate-limiting en&yme involved in arachidonic acid metabolism, thereby generating prostaglandins and thromboxanes which play important roles in supporting and sustaining the inflammatory response 3594. -n rodents as well as in humans, cerebral ischemia upregulated "/-' expression in neurons, blood vessels, and inflammatory cells in the injured brain 3*5,5?, (+4. Moreover, administration of the selective "/-' inhibitor 0$5?9 attenuated the elevation of 1;6' and reduced the infarct in a model of M !" 3*54. "/-' reaction products may also contribute to 0M%!-induced neuronal injury and the pathogenesis of nitric oxide after ischemia 3(*, ('4. 0itric oxide (0"# is an important mediator in the cerebral ischemic injury 3(54. $pecifically, 0itric oxide derived from the inducible isoform (i0"$# expressed by many cells is very important in excitotoxic injury cascades 3*9, *?4. 1harmacologically selective inhibitors of i0"$ attenuated infarct volume after focal cerebral ischemia 3'*,((, ()4. 0itric oxide produced by i0"$ has been shown to contribute to "/-' activity (possibly without altering "/-' expression# 3*:4. -nhibition of i0"$ could also serve as neuroprotection through "/-' inhibition just before the start of the delayed death of !* neurons 3(84. Oe confirmed that cortex tissue obtained from rats with ' hours of M !" followed '( hours reperfusion exhibited significantly more "/-' and i0"$ protein expressions than that of sham group, which supported the idea that inflammatory molecules participate in the occurrence and development of cerebral ischemia. !t the same time, we found that theaflavin-treatment dose dependently inhibited "/-' and i0"$ protein expressions. -n order to elucidate the mechanism of theaflavin on inflammation-related events, we investigated the m@0! expression of "/-' and i0"$ in cerebral ischemic tissues of rats and determined the influence of theaflavin-treatment on m@0! production of "/-' and i0"$. Oe found that the m@0! expressions of "/-' and i0"$ were in accordance with the results of immunohistochemistry detection. @T-1 @ analysis revealed that the m@0! levels of "/-' and i0"$ increased in

brain tissues of the vehicle-treated group. $imilarly, theaflavin had a dose-dependent effect on decreasing m@0! expressions of "/-' and i0"$. This prompted us to investigate the regulation of "/-' and i0"$ gene transcriptions in the process of inflammatory responses. Many cytokines such as -7-8, -7-**, and inflammatory mediators produced by ischemic brain cells, play important roles contributing to ischemic pathophysiology 3(:, (94. A!B-$T!T is an important downstream signal pathway of these cytokines 3(?4. Dinding of neurokines to the membrane receptor leads to dimeri&ation of gp*5+, followed by activation of A!B, which in turn phosphorylates cytoplasmic $T!T. 1hosphorylated $T!T forms homo- or heterodimers and translocates into the nucleus, stimulating gene transcription. Therefore, the A!B-$T!T pathway provides cells with a vital mechanism for responding to various extracellular stimuli including ischemic stress. !ccumulation in the nucleus of tyrosine phosphorylated $T!T dimers is followed by %0! binding, activation of target gene transcription, dephosphorylation, and returns to the cytoplasm 3)+4. $T!T-* induces expression of the transcription factor -@C-*, which then itself binds to specific %0! elements of the i0"$ promoter to further promote i0"$ expression 3)*4. 1retreatment with the Aanus tyrosine kinase (A!B# inhibitor !;-(?+ before the six occlusion-reperfusion cycles blocked both the tyrosine phosphorylation of $T!T*,5 and the subse=uent upregulation of "/-' protein, demonstrating a necessary role of the A!B-$T!T pathway in the induction of "/-' 3)'4. Oe therefore investigated the effect of theaflavin on tyrosine phosphorylation of $T!T-*. "ur results have shown that the levels of $T!T-* phosphorylation on tyrosine :+* were markedly enhanced in brains subjected to ' h of M !" followed by '( hours reperfusion. Theaflavintreatment dose dependently inhibited phosphorylation of $T!T-* and m@0! expressions ( "/-' and i0"$# controlled by it. -n conclusion, our study demonstrated that theaflavin significantly protected neurons from cerebral ischemia-reperfusion injury by limiting lipid peroxidation, leukocyte infiltration and expression of - !M-*. Theaflavin also suppressed upregulations of inflammatory-related prooxidative en&ymes (i0"$ and "/-'# in ischemic brain via, at least in part, reducing $T!T-* phosphorylation. !s a potent antioxidative drug, theaflavin could be beneficial for the prevention and,or amelioration of cerebral ischemia-reperfusion injury. Thus, the protection of neurons by theaflavin may provide clinically beneficial outcomes alone or in combination with thrombolytic therapy. ACKNOWLEDGMENT Oe thank %octor Vao 7iu for checking the spelling of this manuscript.

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