Cardovascuar Scence

3010 MSC
Ischemc Precondtonng
of the myocardum
Contents Page
1.0 Introducton
2.0 Methods and Materas
3.0 Resuts
4.0 Dscusson
5.0 References
6.0 Appendx
1.0 Introduction
Accordng to the Word Heath Organzaton, schemc heart dsease s the
eadng cause of death n westernzed countres and s typcay
charactersed by nsuffcent bood fow to regons of the myocardum,
whch eads to myocarda nfarcton (Downey, Davs, & Cohen, 2007)
(WHO, 2008). It s we recognzed that myocarda schema s a compex
phenomenon affectng the mechanca, eectrca, structura and
bochemca propertes of the heart. Despte ths compexty mpressve
progress has been acheved n advancng our understandng and
apprecaton of the ceuar processes and the mechanstc bases
underyng cardac dysfuncton, assocated wth schema and most
mportanty n appyng ths knowedge to deveop therapeutc
nterventons (Karmayzan, 1996).
The myocardum s crtcay dependent on ts bood suppy, to fuf ts
pumpng requrements. Interrupton of coronary artery bood fow, most
commony a resut of atheroscerotc heart dsease, can ect onc,
eectrophysoogca and contracte dsturbances that mpede or
utmatey suppress the hearts norma functon resutng n myocarda
nfarct (Naran|an, Nobaukra, Man|eet, & Anton, 2003). Obvousy the
hemodynamc determnants of oxygen demand w aso nfuence the
outcome of the schemc nsut and ncude heart rate, wa stress and
myocarda contractty (The McGraw-H Companes, 2008).
Durng acute myocarda nfarcton, the centra area of necross s
generay surrounded by an area of n|ury, whch n turn s surrounded by
an area of schema. Thus, varous stages of myocarda damage can
coexst. The dstncton between schema and necross s whether the
phenomenon s reversbe (AHA, 2008).
Ironcay schema can exert paradoxca effects on the heart, whch are
prmary dependent on the duraton of the schemc nsut. The
myocardum s remarkaby senstve to oxygen deprvaton, thus
proonged perods of schema are vsby detrmenta, resutng n the
geness of etha arrhythmas, contracte dysfuncton and utmatey
necrotc ce death (Levy & Pappano, 2007). Bref perods of schemc
nsuts on the other hand actuay nvoke an ntrnsc protectve
mechansm, whch renders the myocardum more resstant to subsequent
potentay etha schemc epsodes (Dekker, 1998). Ths nnate
physoogca adaptve phenomenon arose from the poneerng work of
Murry, |ennngs and Remer n 1986, who termed ths concept schemc
precondtonng. Ischemc precondtonng was orgnay defned as the

enhancement of myocarda toerance aganst nfarcton nduced

by a bref
subetha epsode of schema n expermenta anmas

(Murry, |ennngs, &
Remer, 1986).

Subsequenty, the concept of schemc precondtonng was
expanded and currenty

ncudes protecton aganst myocarda stunnng,
arrhythma,

and vascuar dysfuncton after schema/reperfuson n|ury
(Tetsu|, 2006).
Ischemc precondtonng nvokes a bphasc pattern of protecton n the
myocardum, an eary or frst wndow of protecton and a deayed or
second wndow of protecton (Ghosh, Ng, Tawar, Squre, & Gananes,
2000) (fgure 1). Murry and coeagues frst descrbed the acutey
manfested phase of adapton. Ths frst wndow of protecton, now
referred to as cassca precondtonng s manfested mmedatey
foowng bref subetha schema and confers a marked sowng n the
progresson of rreversbe schemc n|ury durng subsequent schemc
exposure (Naran|an, Nobaukra, Man|eet, & Anton, 2003). One of the most
dstnctve features of cassca precondtonng s that the protecton
afforded by antecedent schema wanes rapdy. No protecton aganst
ether nfarcton or arrhythmas s observed when the nterva between
precondtonng and the subsequent schemc nsut s extended beyond 2
hours (Baxtor, 1998). The ate phase or second wndow of protecton s
exhbted approxmatey 12 to 72 hours after the nta stmuus and s
charactersed by both nfarct sze reducton and an attenuaton of post
schemc contracte dysfuncton (Przykenk, 2003) Both phases of
precondtonng nvove reducton of necrotc tssue mass (nfarct sze),
mprovement of cardac performance and reducton of arrhythmas
foowng schaema and reperfuson (Mer & Van Wnke, 1999) .
Figure 1: The bphasc phase of protecton nvoked by schemc
precondtonng.
The underyng mechansms of schemc precondtonng have been
extensvey nvestgated, however the bass of such cardoprotecton s not
fuy eucdated. Current evdence suggests that the cardoprotectve
mechansm produced as a resut of schemc precondtonng s a mut-
factora process consstng of an eary nta trgger, an ntermedate
medator and an end effector. (Naran|an, Nobaukra, Man|eet, & Anton,
2003), (Vten-|ohansen, Zhao, |ang, Zatta, & Dobsen, 2007). The most
favoured current hypothess for the frst wndow of precondtonng
suggests that a varety of endogenous gands such as adenosne,
bradyknn, catechoamnes and opods actvate G-proten couped
receptors nked to proten knase C (PKC) to ntate an ntraceuar
transducton pathway (Downey, Davs, & Cohen, 2007). It s thought PKC
may actvate a tyrosne knase, whch n turn actvates MAP knase eadng
fnay to the phosphoryaton of effector protens, and possby the
actvaton of mtochondra ATP senstve potassum channes (mtoK
+
ATP
)
(Przykenk, 2003)(Ghosh, Ng, Tawar, Squre, & Gananes, 2000). An
aternatve canddate for ntatng schemc precondtonng

s a
preschemc md ncrease of the ntraceuar Ca
2+
concentraton

(|Ca
2+
|).
Short perods of schema and reperfuson nduce

a oss of Ca
2+
from the
sarcopasmc retcuum (SR) and an accumuaton

of Ca
2+
n the cytoso
(Wu & Feher, 1995). Myawak and

co-workers nferred that a sma
ncrease of |Ca
2+
| acts as

a medator of precondtonng va actvaton of
PKC (Myawak & Ashraf, 1997)

These studes provde nove nformaton to understand the underyng
mechansm of protecton by precondtonng of the myocardum and the
resuts have obvous cnca mportance.
The purpose of ths study s to nvestgate the degree to whch schemc
precondtonng renders the myocardum more resstant to subsequent
subetha schemc epsodes, by measurng the return of functon, of
systoc, dastoc and deveoped pressure as we as fow rate n
precondtoned hearts versus contro hearts.
2.0 Methods
Mouse Heart Perfusion Model
Precondtoned or genetcay modfed C57/B6 mce were anesthetzed
wth 50 mg/kg of sodum pentobarbtone admnstered ntra-pertoneay. A
thoracotomy was performed and hearts rapdy excsed nto ce-cod Krebs
bcarbonate buffer. The aorta was then cannuated and hearts perfused n
a Langendorff mode at a pressure of 80 mmHg, as outned by us n deta
prevousy. Hearts were perfused wth a modfed Krebs bcarbonate buffer
contanng (n mM): NaC, 118; NaHCO
3
, 25; KC, 4.7; KH
2
PO
4
, 1.2; CaC
2
,
2.5; MgSO
4
, 1.2; EDTA, 0.6; and wth 11 mM gucose as exogenous
substrates. The perfuson fud was saturated wth 95% O
2
and 5% CO
2
at
37C (yedng a pH of 7.4 and a PO
2
>550 mmHg at the aortc cannua),
and was contnuousy ftered va a 0.45 n-ne fter. The eft ventrce
was vented wth a poyethyene dran to prevent fud accumuaton and an
ntra-ventrcuar baoon ntroduced for assessment of contracte functon,
as descrbed prevousy. Coronary fow was montored va an utrasonc
fow-probe n the aortc perfuson ne connected to a T106 fowmeter
(Transonc Systems Inc, Ithaca, NY, USA). Functona data were recorded at
1 KHz on a 4 channe MacLab data acquston system (ADInstruments,
Caste H, Austraa). Ventrcuar pressure was dgtay processed to yed
systoc and dastoc pressures and heart rate.
Ischemia-reperfusion protocol. After nstrumentaton a hearts were
ntroduced nto a water-|acketed chamber superfused wth warmed
buffer at 37C, ensurng stabe temperature throughout the
protocos. Foowng a 20 mn stabzaton perod at ntrnsc heart
rate hearts were swtched to eectrca pacng at 420 beats/mn

and
stabzed a further 10 mn. Hearts were excuded from anayss at
ths tme f they met one of the foowng crtera: ) eft ventrcuar
systoc pressure <100 mmHg; ) coronary fow >5 m/mn; )
fuctuatng (unstabe) contracte functon; or v) sgnfcant
arrhythma's. Basene functona measurements were made before
sub|ectng hearts to 20 mn goba normothermc schema and 45-
60 mn of reperfuson.
3.0 Results
The early protective effect of ischemic preconditioning
and comparison of its efficacy ith control hearts! in the
first indo of preconditioning.
As shown n fgure 2, after 20 mnutes of schema a marked ncrease n
dastoc pressure was observed n the contro hearts. The precondtoned
hearts on the other hand showed an nta rse n dastoc pressure durng
the schemc perod, however once reperfused the dastoc pressure
dropped back down to basene eve. The tra|ectory of recovery of
dastoc pressure s sgnfcanty mproved n the precondtoned hearts
reatve to contro P<0.05.
Figure 2: Demonstrates the effect of schemc precondtonng on the
tme-course and the post schemc recovery of dastoc
pressure on adut mce hearts. The dfferences n dastoc
pressure between contro and precondtoned
P vaue =
0.021012
hearts were measured over a 20 mnute perod of
schema, foowed subsequenty by a 60 mnute perod of
reperfuson. Each pont represents a mean S.E.M for 4
hearts. Note that the tra|ectory of recovery of
dastoc pressure s sgnfcanty mproved by schemc
precondtonng reatve to contro *(P vaue = 0.021012)
As shown n fgure 3, after the 20 mnute schemc perod the recovery of
systoc pressure mproved wth the precondtoned hearts, however ths
recovery does not show a sgnfcant dfference n comparson to the
contro hearts P>0.05.
Figure 3: Demonstrates the effect of schemc precondtonng on the
tme-course and the post schemc recovery of systoc
pressure on adut mce hearts. The dfferences n
systoc pressure between contro and precondtoned
hearts were measured over a 20 mnute perod of
P vaue =
0.19479
schema, foowed subsequenty by a 60 mnute perod of
reperfuson. Each pont represents a mean S.E.M for 4
hearts. Evdenty the recovery of systoc pressure after a perod
of schema s mproved wth schemc precondtonng,
however ths recovery does not show a sgnfcant dfference n
comparson to the contro hearts (P vaue = 0.19479).
It s evdent from fgure 4, that schemc precondtonng has a profound
effect on the recovery of deveoped pressure foowng the 20 mnute
schemc perod. In comparng contro vaues wth precondtoned vaues
the recovery of deveoped pressure was found to be sgnfcanty
ncreased wth schemc precondtonng P<0.05.
Figure ": Demonstrates the effect of schemc precondtonng on the
tme-course and the post schemc recovery of deveoped
pressure on adut mce hearts. The dfferences n deveoped
pressure between contro and precondtoned hearts
were measured over a 20 mnute perod of
schema, foowed subsequenty by a 60 mnute perod of
reperfuson. Each pont represents a mean S.E.M for 4
hearts. Notceaby the recovery of deveoped pressure after a
perod of schema mproved consderaby wth schemc
precondtonng n comparson to the contro hearts.
(P vaue = 0.016495).
P vaue =
As shown n fgure 5 schemc precondtonng appeared to have tte
effect on the recovery of fow durng the post the schemc perod. The
dfference n recovery between contro and precondtoned hearts was
found to be nsgnfcant P>0.05.
Figure #: Demonstrates the effect of schemc precondtonng on the
tme-course and the post schemc recovery of fow rate n
adut mce hearts. The dfferences n fow rate between contro
and precondtoned hearts were measured over a 20 mnute
perod of schema, foowed subsequenty by a 60 mnute
perod of reperfuson. Each pont represents a mean S.E.M
for 4 hearts. The recovery of fow rate after a perod of
schema was actuay ower wth schemc precondtonng n
comparson to the contro hearts, however there was no
sgnfcant dfference (P vaue = 0.137706).
P vaue =
".0 $iscussion
It has prevousy been demonstrated by Murry, |ennngs and Remer, that
a bref epsode of schema sows the rate of ATP depeton durng
subsequent schemc epsodes (Murry, |ennngs, & Remer, 1986).
Addtonay, ntermttent reperfuson may be benefca to the
myocardum by washng out catabotes that have accumuated durng
schema (Baxtor, 1998). Thus, t s proposed that mutpe bref schemc
epsodes mght actuay render the heart more resstant to a subsequent
sustaned schemc nsut. To test ths hypothess, the foowng
experment was performed. One group of adut mce (n = 4) were
precondtoned wth three, 1 mnute crcumfex occusons, each separated
by 5 mnutes of reperfuson, foowed by a sustaned 20 mnute occuson
and subsequenty reperfused for 60 mnutes. The contro group (n = 4)
receved a snge 20 mnute occuson foowed subsequenty by a 60
mnute perod of reperfuson. The dastoc pressure, systoc pressure,
deveoped pressure and fow rate of each of the hearts were measured
over the 80 mnute schema/ reperfuson perod are presented n
appendx 1. The effects of schemc precondtonng were determned by
measurng post schemc return of functon as an end pont.
%ffect of ischemic preconditioning on diastolic pressure
In the 80 mnute study, precondtonng wth schema paradoxcay
reduced dastoc pressure to 38.8% of that seen n the contro group. The
mean dastoc pressure vaue at the end of the 60 mnute reperfuson
perod for the precondtoned hearts was 0.14mmHg, ndcatng a 100%
post schemc return of functon, 62.2% greater than that experenced by
the contro hearts. These resuts returned a P vaue of 0.021012,
ndcatng a sgnfcant dfference between the two groups n regards to
the return of functon.
%ffect of ischemic preconditioning on systolic pressure
It can be seen from fgure 3 that the recovery of systoc pressure after a
perod of schema s margnay mproved wth schemc precondtonng.
The extent to whch schemc precondtonng ncreases post-schemc
return of functon s however mted, as the resuts of our experment
ndcate that the precondtoned hearts exhbt ony a 2% ncrease of post-
schemc return of functon over the contro hearts. These resuts returned
a P vaue of 0.19479, ndcatng that the protecton conferred by schemc
precondtonng s reatvey nsgnfcant wth regards to return of functon
of systoc pressure.
%ffect of ischemic preconditioning on developed pressure
As shown n fgure 4, the tra|ectory of recovery of deveoped pressure s
sgnfcanty mproved by schemc precondtonng reatve to the contro
(P vaue = 0.021012). The mean deveoped pressure vaue at the end of
the 60 mnute reperfuson perod for the precondtoned hearts was
109.11mmHg, ndcatng a 61.5% post-schemc return of functon, 14.2%
greater than that experenced by the contro hearts.
%ffects of ischemic preconditioning on flo rates
The expermenta resuts obtaned for the effects of schemc
precondtonng on fow rates were somewhat unexpected. One woud
assume that because schemc precondtonng has resuted n an
ncreased post-schemc return of functon wth a of the prevous end
pont measures, smar resuts coud be expected wth regard to fow rate.
Ths however was not the case as the resuts n fgure 5 shows that the
contro hearts demonstrated a greater return of functon then the
precondtoned hearts. The mean fow rate vaue at the end of the 60
mnute reperfuson perod for the precondtoned hearts was
18.51m/mn/g ndcatng a 61.5% post schemc return of functon, 6.5%
ower than that experenced by the contro hearts. These resuts are
contradctory to the resuts obtaned from severa other studes, ncudng
those done by Morrs and co-workers who reported an ncreased post-
schemc return of functon n precondtoned hearts n comparson to
contro hearts (Karmayzan, 1996).
Taken as a whoe the resuts from ths study strongy suggest that
schemc precondtonng provdes an extremey potent endogenous
cardoprotectve mechansm, whch renders the myocardum more
resstant to subsequent potentay etha perods of schema. The
mechansm by whch schemc precondtonng confers protecton st
remans under ntense nvestgaton, however studes done so far have
found that schemc precondtonng s mut-factora process consstng of
severa key nta trggers, whch actvate ntermedate medators whch
subsequenty converge upon an end effector.
The mechanism of ischemic preconditioning
Athough not fuy eucdated the trgger mechansm of schemc
precondtonng has been found to nvove a number of endogenousy
reeased substances such as adenosne, bradyknn and opods, whch are
reeased durng schema and act n parae to trgger the precondtoned
state through actvaton of mutpe G-couped proten receptors (Downey,
Davs, & Cohen, 2007). Ths mutpe trgger theory proposed by Goto and
coeagues requres that a trggers converge upon a common target. It
was found that protecton afforded by a of the trgger substances coud
be bocked by PKC nhbtors and thus PKC s thought to be ths common
target (Goto, Lu, Yang, Arde, M, & Downey, 1995) (Mk, Cohen, &
Downey, 1998). Whe adenosne coupes drecty to PKC through
phosphopases, bradyknn and opods do so through a compex pathway,
nvovng epderma growth factor receptors, phosphatdynosto 3-
knase, Akt, ntrc oxde synthase, guanyy cycse and PKG (Fgure 6).
Pan et a, proposed that receptor stmuaton ed to the openng of
mtochondra ATP senstve potassum channes, whch resuts n an nfux
of potassum that causes the mtochondra to swe whch somehow ead
to the producton of oxygen based free radcas, whch n turn actvate
PKC (Pan, Yang, & Crtz, 2000). PKC s the centra medator of schemc
precondtonng, actvatng the survva knases Akt and ERK whch are
thought to phosphoryate GSK-3, whch n turn nhbts the formaton of
mtochondra permeabty transton pores n eary reperfuson, thus
conferrng protecton (Downey, Davs, & Cohen, 2007).
Figure &: The proposed mechansm by whch schemc precondtonng
confers protecton (Downey, Davs, & Cohen, 2007).
Athough there s currenty no cncay proven therapes that are wdey
used to enhance the myocardums toerance to schema and thereby
extend the tme-wndow for tssue savage by reperfuson, therapes such
as schemc precondtonng woud be expected to be of vaue not ony n
the management of acute myocarda nfarcton but aso n cardac
surgery, where the heart s often sub|ected to perods of goba schema
and reperfuson. In the surgca settng, schemc precondtonng can be
used n con|uncton wth exstng cardoprotectve strateges, such as
cardopegc arrest or ntermttent ventrcuar fbraton, to enhance post
schemc myocarda vabty and functon.
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protects the human myocardum from schemc n|ury. Oxford ournals ! "# (3),
440-447.
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ryanodne bndng and Ca2+ uptake of cardac sarcopasmc retcuum. $ol Cell
Cardiol , 1965-1975.
&.0 'ppendi(
)ontrol )onditions Time *min+ $iastolic ,ystolic $eveloped
Flo
*ml-min-g+
1 Normoxa 0 3.0 163.4 174.0 35.2
Ischaema 5 51.0 0.0 0.0 0.0
Ischaema 10 84.4 0.0 0.0 0.0
Ischaema 15 71.4 0.0 0.0 0.0
Ischaema 20 59.4 0.0 0.0 0.0
Reperfuson 22 18.7 72.4 53.7 32.8
Reperfuson 25 22.6 52.7 30.1 33.8
Reperfuson 30 30.2 50.7 20.4 30.8
Reperfuson 35 41.5 57.2 15.7 25.5
Reperfuson 40 44.8 67.8 23.0 23.8
Reperfuson 45 39.6 83.6 44.0 25.0
Reperfuson 50 32.0 105.3 73.3 25.7
Reperfuson 55 29.8 95.6 65.9 25.6
Reperfuson 60 27.1 98.2 71.1 25.0
Reperfuson 65 24.5 102.1 77.6 24.3
Reperfuson 80 24.7 99.6 74.9 23.9
2 Normoxa 0 2.0 147.9 149.8 26.0
Ischaema 5 24.5 0.0 0.0 0.0
Ischaema 10 72.1 0.0 0.0 0.0
Ischaema 15 57.5 0.0 0.0 0.0
Ischaema 20 47.8 0.0 0.0 0.0
Reperfuson 22 17.5 69.5 51.9 28.1
Reperfuson 25 21.1 79.1 58.0 31.2
Reperfuson 30 25.4 43.3 17.8 30.3
Reperfuson 35 25.4 79.4 54.0 25.6
Reperfuson 40 24.8 77.1 52.2 22.7
Reperfuson 45 20.5 96.4 76.0 22.2
Reperfuson 50 21.4 89.0 67.6 24.1
Reperfuson 55 20.2 92.3 72.1 22.3
Reperfuson 60 18.6 96.9 78.3 20.4
Reperfuson 65 14.3 97.8 83.6 18.9
Reperfuson 80 14.1 91.6 77.5 16.9
3 Normoxa 0 4.0 144.6 151.2 29.4
Ischaema 5 20.6 0.0 0.0 0.0
Ischaema 10 84.8 0.0 0.0 0.0
Ischaema 15 70.1 0.0 0.0 0.0
Ischaema 20 59.2 0.0 0.0 0.0
Reperfuson 22 17.5 75.9 58.5 31.8
Reperfuson 25 28.8 46.6 17.8 36.2
Reperfuson 30 32.3 36.8 4.5 34.4
Reperfuson 35 37.1 56.4 19.3 30.2
Reperfuson 40 41.0 81.5 40.5 27.5
Reperfuson 45 33.7 81.8 48.2 27.7
Reperfuson 50 30.1 80.6 50.5 28.0
Reperfuson 55 24.0 93.4 69.4 27.7
Reperfuson 60 22.6 88.5 65.9 27.3
Reperfuson 65 21.2 90.9 69.7 26.9
Reperfuson 80 19.3 89.9 70.6 26.5
4 Normoxa 0 1.0 137.2 146.2 33.9
Ischaema 5 72.5 0.0 0.0 0.0
Ischaema 10 84.6 0.0 0.0 0.0
Ischaema 15 72.3 0.0 0.0 0.0
Ischaema 20 62.1 0.0 0.0 0.0
Reperfuson 22 25.7 75.8 50.1 31.0
Reperfuson 25 30.6 56.7 26.0 32.5
Reperfuson 30 29.4 45.3 16.0 31.8
Reperfuson 35 34.6 58.9 24.3 28.5
Reperfuson 40 38.8 72.6 33.8 26.4
Reperfuson 45 38.6 68.0 29.4 26.4
Reperfuson 50 27.3 72.8 45.5 27.3
Reperfuson 55 23.9 74.4 50.4 27.6
Reperfuson 60 20.4 79.6 59.2 27.0
Reperfuson 65 17.2 86.8 69.6 26.3
Reperfuson 80 16.3 87.1 70.9 25.5
Ischaemic
.reconditi
oning Condtons Tme (mn) Dastoc Systoc Deveoped
Fow
(m/mn/g)
1 Normoxa 0 3.0 167.8 172.6 26.9
Ischaema 5 -3.5 0.0 0.0 0.0
Ischaema 10 48.1 0.0 0.0 0.0
Ischaema 15 65.2 0.0 0.0 0.0
Ischaema 20 61.0 0.0 0.0 0.0
Reperfuson 22 11.1 123.2 112.1 29.0
Reperfuson 25 5.6 125.2 119.6 34.2
Reperfuson 30 19.7 82.2 62.5 30.9
Reperfuson 35 13.8 73.5 59.7 21.9
Reperfuson 40 6.7 73.4 66.6 18.5
Reperfuson 45 3.0 82.2 79.3 18.0
Reperfuson 50 0.7 88.5 87.8 17.3
Reperfuson 55 -1.3 92.7 94.0 16.2
Reperfuson 60 -3.1 100.1 103.1 15.5
Reperfuson 65 -3.9 101.3 105.2 14.8
Reperfuson 80 -4.2 103.1 107.4 14.6
2 Normoxa 0 5.0 157.0 165.1 26.5
Ischaema 5 -3.4 0.0 0.0 0.0
Ischaema 10 64.4 0.0 0.0 0.0
Ischaema 15 64.1 0.0 0.0 0.0
Ischaema 20 60.4 0.0 0.0 0.0
Reperfuson 22 8.4 154.0 145.6 34.9
Reperfuson 25 9.0 153.4 144.5 35.2
Reperfuson 30 14.1 77.7 63.7 33.8
Reperfuson 35 7.2 71.0 63.8 24.4
Reperfuson 40 2.8 71.2 68.4 21.2
Reperfuson 45 0.3 77.7 77.4 20.0
Reperfuson 50 -2.0 84.9 86.8 19.3
Reperfuson 55 -3.5 88.7 92.2 18.1
Reperfuson 60 -4.6 91.8 96.4 17.7
Reperfuson 65 -4.8 89.8 94.6 16.1
Reperfuson 80 -5.1 90.1 95.1 15.3
3 Normoxa 0 2.0 182.7 194.1 40.7
Ischaema 5 1.4 0.0 0.0 0.0
Ischaema 10 99.9 0.0 0.0 0.0
Ischaema 15 87.7 0.0 0.0 0.0
Ischaema 20 78.7 0.0 0.0 0.0
Reperfuson 22 29.2 91.3 62.1 30.2
Reperfuson 25 34.0 79.4 45.4 34.4
Reperfuson 30 41.3 48.1 6.8 29.9
Reperfuson 35 31.2 85.9 54.7 28.6
Reperfuson 40 24.0 105.7 81.6 28.1
Reperfuson 45 19.8 119.0 99.2 28.3
Reperfuson 50 18.0 123.7 105.7 28.2
Reperfuson 55 15.9 127.7 111.9 27.7
Reperfuson 60 13.5 131.9 118.5 27.4
Reperfuson 65 11.5 132.7 121.1 26.8
Reperfuson 80 10.5 133.8 123.2 26.6
4 Normoxa 0 0.0 169.5 177.3 26.2
Ischaema 5 -2.8 0.0 0.0 0.0
Ischaema 10 41.9 0.0 0.0 0.0
Ischaema 15 65.8 0.0 0.0 0.0
Ischaema 20 65.2 0.0 0.0 0.0
Reperfuson 22 13.1 123.8 110.7 36.0
Reperfuson 25 19.0 65.1 46.1 36.7
Reperfuson 30 21.6 69.7 48.1 33.2
Reperfuson 35 10.5 88.4 77.8 22.8
Reperfuson 40 6.3 88.7 82.4 19.3
Reperfuson 45 2.5 100.1 97.5 19.1
Reperfuson 50 0.6 106.1 105.5 18.8
Reperfuson 55 0.8 103.7 102.9 18.5
Reperfuson 60 0.4 107.8 107.4 18.2
Reperfuson 65 -0.1 108.6 108.8 17.8
Reperfuson 80 -0.7 110.0 110.7 17.5