Asiatic Acid Attenuates Infarct Volume, Mitochondrial
Dysfunction, and Matrix Metalloproteinase-9 Induction
After Focal Cerebral Ischemia
Ki Yong Lee, PhD; Ok-Nam Bae, PhD; Kelsey Serfozo; Siamk Hejabian; Ahmad Moussa;
Mathew Reeves, PhD; Wilson Rumbeiha, PhD; Scott D. Fitzgerald, DVM, PhD; Gary Stein, PharmD;
Seung-Hoon Baek, PhD; John Goudreau, DO, PhD; Mounzer Kassab, MD; Arshad Majid, MD
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Background and Purpose—Asiatic acid (AA) has been shown to attenuate cerebral infarction in a mouse model of focal
ischemia and shows promise as a neuroprotective stroke therapy. To facilitate translation of these findings to clinical
studies, we determined pharmacokinetics, a dose–response relationship, the therapeutic time window, and efficacy using
multiple stroke models. We also explored potential mechanisms of action.
Methods—Escalating doses of intravenous AA were administered and serum concentrations were measured at multiple
time points for the pharmacokinetic studies. Subsequently, a dose–response relationship was determined followed by
administration at different intervals after the onset of ischemia to establish a therapeutic time window for
neuroprotection. Outcome measurements included both histological and behavioral. Mitochondrial function and matrix
metalloproteinase activity in controls and treated rats were also determined.
Results—The pharmacokinetic studies showed that AA (75 mg/kg) has a half-life of 2.0 hours. AA significantly decreased
infarct volume and improved neurological outcome even when administration was at time points up to 12 hours after
the onset of ischemia. Infarct volume was also significantly decreased in female rats and spontaneously hypertensive
rats. AA attenuated mitochondrial dysfunction and reduced matrix metalloproteinase-9 induction.
Conclusions—Our study shows AA is effective against multiple models of focal ischemia, has a long therapeutic time
window, and is also effective in females and hypertensive animals. AA may mediate neuroprotection by protecting
mitochondria and inhibiting matrix metalloproteinase-9 induction and activation. Taken together these data suggest that
AA is an excellent candidate for development as a stroke therapy. (Stroke. 2012;43:00-00.)
Key Words: asiatic acid 䡲 matrix metalloproteinase-9 䡲 mitochondria 䡲 neuroprotective 䡲 stroke
S
troke is a leading cause of mortality and the largest single
cause of adult disability worldwide.1 Treatment with
tissue-type plasminogen activator, the only approved pharmacological therapy, is limited by its narrow time window and
the risk of hemorrhagic complications. A plethora of neuroprotectants have failed to translate to clinically effective
therapies and a desperate need exists for new therapies.2
Asiatic acid (AA) is a triterpene isolated from Centella
asiatica, which has been widely used as an antioxidant and
anti-inflammatory agent.3 AA has also been shown to display
robust neuroprotective properties both in vitro and in vivo
and shows promise as a novel therapeutic agent for acute
stroke therapy.4 –7
The Stroke Therapy Academic Industry Roundtable
(STAIR)8 has published recommendations for preclinical
testing of candidate stroke therapies. These recommendations include determining: a dose–response relationship, a
therapeutic time window, and efficacy in multiple models
including permanent and transient ischemia, female animals, aged animals, and animals with comorbidities. Efficacy in both behavioral and histological time points was
also recommended.
Previously, we reported that AA is neuroprotective in a
mouse model of permanent focal ischemia and protects the
integrity of the blood– brain barrier (BBB) after ischemia.7 In
accord with STAIR guidelines, we have now tested AA in
Received September 28, 2011; final revision received January 7, 2012; accepted January 27, 2012.
From the Division of Cerebrovascular Diseases (K.Y.L., O.B., K.S., S.H., A.M., S.B., J.G., M.K., A.M.), Department of Neurology and
Ophthalmology, Department of Pathobiology and Diagnostic Investigation, and Diagnostic Center for Population & Animal Health (S.D.F.), Department
of Epidemiology (M.R.), and Department of Medicine (G.S.), Michigan State University, East Lansing, MI; the College of Pharmacy (O.B.), Hanyang
University, Ansan, Gyeonggido, Korea; the College of Pharmacy (S.B.), Ajou University, Suwon, Gyeonggido, Korea; Veterinary Diagnostic and
Production Animal Medicine (W.R.), College of Veterinary Medicine, Iowa State University, Ames, IA; and the Department of Neurology (A.M.), Salford
Royal Hospital, Salford, UK.
The online-only Data Supplement is available with this article at http://stroke.ahajournals.org/lookup/suppl/doi:10.1161/STROKEAHA.111.
639427/-/DC1.
Correspondence to Arshad Majid, MD, Department of Neurology, Salford Royal Hospital, Stott Lane, Salford, UK 0161 2064279. E-mail
[email protected]
© 2012 American Heart Association, Inc.
Stroke is available at http://stroke.ahajournals.org
DOI: 10.1161/STROKEAHA.111.639427
1
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June 2012
multiple stroke models using both histological and functional
outcomes and established a dose–response relationship and a
therapeutic time window. We also investigated the mechanism of action and provide evidence of reduced mitochondrial damage and matrix metalloproteinase inhibition.
Materials and Methods
Animals
Adult male Sprague-Dawley (SD), female SD rats, and male spontaneously hypertensive rats (SHR) weighing 250 to 300 g were used
(Harlan, Indianapolis, IN). All experiments were performed in
accord with the National Institutes of Health Policy and Animal
Welfare Act under the approval by Institutional Animal Care and
Use Committee at Michigan State University.
Drugs
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AA was dissolved to a concentration of 50 mg/mL in ubisol-Aqua
(Zymes LLC, Hasbrouck Heights, NJ). The lateral tail vein was used
for the administration of AA or ubisol-Aqua (vehicle).
Blinding and Randomization
Treatment groups were allocated in a randomized fashion using a
Researcher Randomizer (www.randomizer.org/). Investigators were
blind to the allocation of treatment when doing surgeries and doing
outcome evaluations.
Justification for the Ischemia Models Used in
These Studies
There is no ideal stroke model that completely reflects clinical stroke
in humans. We used both permanent (pMCAO) and transient
(tMCAO) middle cerebral artery occlusion models using a filament
to occlude the middle cerebral artery through the carotid (see the
online-only Data Supplement; http://stroke.ahajournals.org). The
pMCAO models the clinical situation of occlusion of the cerebral
vessel without recanalization. The tMCAO models the recanalization
of a previously occluded cerebral vessel either spontaneously or with
treatment.9 Because stroke can involve both situations, we tested
both models.
Experimental Groups
Detailed methods of blood sampling and pharmacokinetics studies,
surgery and induction of ischemia, neurological score, behavioral
testing, calculation of infarct volume using triphenyltetrazolium
chloride and cresyl violet staining, brain mitochondrial isolation,
respiration measurements, in vivo matrix metalloproteinase (MMP)
level measurements, and statistical analyses are available in the
online-only Data Supplement.
Experiment 1: Determination of Pharmacokinetics
and Safety/Tolerability of AA
We sought to determine pharmacokinetics and safety/tolerability of
AA. Male SD rats (9 –10 weeks) were randomly divided into 4
groups (n⫽7 rats/group) and given a single intravenous bolus
injection of vehicle or 10, 25, and 75 mg/kg AA. Blood samples were
drawn before administration and at 15 minutes, 1 hour, 3 hours, 6
hours, 12 hours, and 24 hours postadministration. The concentration
of AA in serum at each time point was measured using highperformance liquid chromatography/mass spectroscopy. Rats were
allowed to survive for 14 days. For safety and tolerability evaluations, the daily assessment occurred of the following: (1) food
consumption; (2) body weight; (3) activity; and (4) mortality. All
animals underwent blood analysis for chemistry and hematology. In
a separate set of experiments, AA or vehicle was administered to rats
(n⫽4 per group) and blood pressure was monitored for 24 hours.
Experiment 2: Determination of a
Dose–Response Relationship
We sought to determine the dose–response relationship and the
therapeutic time window in healthy young males in both pMCAO
and tMCAO models.
Male SD rats were randomly assigned to treatment with AA
(50 –75 mg/kg) or vehicle at 30 minutes (AA 50 mg/kg, n⫽21; AA
75 mg/kg, n⫽23; vehicle, n⫽25) before ischemia or 6 (AA 75
mg/kg, n⫽20; vehicle, n⫽20), 9 (AA 75 mg/kg, n⫽18; vehicle,
n⫽17), and 12 (AA 75 mg/kg, n⫽19; vehicle, n⫽18) hours after
ischemia onset in the pMCAO model.
Similarly, rats were randomly assigned treatment with AA or
vehicle at 6 (AA 75 mg/kg, n⫽18; vehicle, n⫽17), 9 (AA 75 mg/kg,
n⫽18; vehicle, n⫽20), or 12 (AA 75 mg/kg, n⫽19; vehicle, n⫽21)
hours after ischemia onset in the tMCAO model. Infarct volumes
were evaluated at 24 hours after middle cerebral artery occlusion
(MCAO).
Experiment 3: Determination of the Influence of
Gender on Outcomes
Clinical stroke affects both genders. We sought to determine the
influence of gender on outcomes. Female SD rats (9 –10 weeks) were
subjected to pMCAO and treated with AA (75 mg/kg, n⫽11) or
vehicle (n⫽12) at 6 hours after ischemia onset.
Experiment 4: Determination of the Influence of
Hypertension on Outcomes
Clinical stroke affects patients with comorbidities like hypertension.
We sought to determine the efficacy of AA in the presence of
hypertension. SHRs were subjected to tMCAO. Only tMCAO was
used because SHRs have very high mortality rates with pMCAO.10
SHRs were subjected to 1 hour tMCAO and treated with AA (75
mg/kg, n⫽12) or vehicle (n⫽13) at 6 hours after ischemia onset.
Infarct volumes were evaluated at 24 hours after MCAO.
Experiment 5: Determination of Long-Term
Histological and Behavioral Outcomes
We sought to determine long-term histological and behavioral
outcomes. Male SD rats were randomly divided into 3 groups. (Only
the transient model was used because like other investigators, we
also observed high long-term mortality.) The sham group had
surgery but was not subjected to tMCAO (n⫽6). The other 2 groups
were subjected to tMCAO and blindly treated with AA (75 mg/kg,
n⫽23) or vehicle (n⫽18) at 6 hours after ischemia onset. The
neurological scores, behavioral tests, and infarct volumes were
evaluated during and at the end of 2 weeks post-MCAO.
Experiment 6: Exploration of Possible Mechanisms
of Action
We explored possible mechanism of action by examining the effects
of AA on brain mitochondrial bioenergetics and MMP induction and
activity. Both these mechanisms play a role in ischemic injury.11–13
Male SD rats were randomly divided into 2 groups and subjected to
tMCAO and treated with AA (75 mg/kg, n⫽8) or vehicle (n⫽8) at
6 hours after ischemia onset. The brain tissues were isolated at 24
hours after MCAO for mitochondrial isolation and Western blots.
(Details of the tissue isolation are provided in the only-only Data
Supplement).
Results
Safety, Tolerability, and Pharmacokinetics
After a single intravenous injection of 10, 25, and 75 mg/kg,
serum concentrations of AA increased to 2.95, 6.16, and
8.62 ␮mol/L, respectively, at 15 minutes. The subsequent fall
in serum concentrations within the first 1 hour was rapid
(Figure 1). The T1/2 of 75 mg/kg AA was 2.00⫾0.18 hours.
The maximal concentration value and area under the curve
Lee et al
AA Protects Against Focal Cerebral Ischemia
3
Figure 1. Serum concentration of AA after an intravenous injection at 10, 25, and 75 mg/kg. AA indicates asiatic acid.
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value of 75 mg/kg AA was 9.70⫾0.82 mg/L and 27.67⫾2.91
mg 䡠 hr/L, respectively (online-only Data Supplement Table
VI). AA was well tolerated up to a dose of 75 mg/kg. Higher
doses could not be tested because of toxicity associated with
the solvent. Food consumption, activity, weight, mean arterial
pressure, heart rate, and mortality are presented in onlineonly Data Supplement Figures I through V.
AA Attenuates Infarct Volume After
Ischemic Stroke
Dose–Response Relationship and Therapeutic
Time Window
AA administration (50 or 75 mg/kg, 30 minutes before
MCAO) had a dose-dependent protective effect, attenuating
infarct volume against pMCAO at 24 hours by 37.0% and
52.5%, respectively (Figure 2A). To determine the therapeutic time window, rats were administered AA (75 mg/kg) at 6,
9, or 12 hours after onset of ischemia. As illustrated in Figure
2B, treatment with AA significantly decreased infarct volume
even 12 hours after pMCAO. In tMCAO, infarct volume was
significantly reduced with AA treatment at 6 or 9 hours,
53.1% and 52.3%, respectively (Figure 2C).
Efficacy in Female Rats and Hypertensive Rats
Additional studies were performed in female animals and
hypertensive animals. As illustrated in Figure 3A, 6 hours
posttreatment with AA (75 mg/kg) in female rats decreased
infarct volume by 56.4%. Similarly, AA (75 mg/kg) 6 hours
postischemia in SHR significantly reduced infarct volume by
25.6% (Figure 3B).
Determination of Long-Term Efficacy of Histological and
Functional Outcomes
To test whether the neuroprotective effect of AA is sustained,
AA (75 mg/kg) was administered 6 hours after onset of
ischemia. Neurological function and cerebral infarct volume
were determined at 14 days. AA reduced infarct volume by
36.3% (Figure 4A). AA treatment also significantly improved
functional outcome and decreased motor, sensory, and reflex
impairment 1, 3, 7, and 14 days after ischemia (Figure 4B).
Additional functional testing using the adhesive removal and
Rotorod tests were done before (baseline) and 1, 3, 7, and 14
days after ischemia (Figure 4C–D). In the Rotorod test, a
strong trend toward better performance of the group injected
Figure 2. Protective effect of AA on infarct volume. AA was administrated 30 minutes before ischemia onset in pMCAO (A). AA treatments (50 mg/kg and 75 mg/kg) significantly reduced infarct volume compared with the vehicle group. AA (75 mg/kg) was
administrated at 6, 9, or 12 hours after ischemia onset in pMCAO
(B) and tMCAO (C) models. Infarct volume was determined at 24
hours after MCAO by TTC staining. AA treatment significantly
reduced infarct volume up to 12 hours postischemia onset. Scale
bar, 1 cm. All values are means⫾SEM and analyzed by 1-way
analysis of variance. *P⬍0.05, **P⬍0.01, ***P⬍0.001 versus vehicle
group. AA indicates asiatic acid; pMCAO, permanent middle cerebral artery occlusion; tMCAO, transient middle cerebral artery
occlusion; TTC, triphenyltetrazolium chloride.
with AA was observed at postoperative Day 14 (repeatedmeasures analysis of variance, P⫽0.056). Significant improvement for the adhesive removal test was observed at
postoperative Day 14 (P⫽0.027; Figure 4D).
AA Influences Mitochondrial Health and MMPs
Influence of AA on Mitochondrial Respiration Functions
Brain mitochondrial bioenergetics were characterized by
measuring oxygen consumption rates of different substrates
and calculating the respiratory control ratio (RCR; Figure 5).
The measured rates of oxygen consumption of different
mitochondrial substrates and the derived RCR serve as
important indicators of mitochondrial respiratory capacity
and functional homeostasis.11,12 Adenosine 5⬘-triphosphate
phosphorylation (State III) rates were significantly decreased
in mitochondria sampled from the ipsilateral hemisphere after
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June 2012
Western blotting. Ischemic rats showed an increase in
MMP-9 levels in the ipsilateral hemisphere, whereas no
changes in MMP-2 were detected (Figure 6A–B). AA treatment significantly inhibited MMP-9 induction (Figure 6C).
To determine whether AA inhibits MMP activity, an in
vitro assay using recombinant MMP-9 was performed. The
MMP-9 was incubated with various concentrations (0.1, 0.5,
and 1 mmol/L) of AA, and the effect on enzyme activity was
determined by the ability to degrade fluorescently labeled
gelatin. AA significantly inhibited MMP-9 (Figure 6D) in a
concentration-dependent manner. Because MMPs are zincdependent endopeptidases, AA inhibitory activity in excess
zinc was examined to determine whether AA has a metalchelating effect on zinc. Our results suggest that AA does not
inhibit MMP activity by zinc chelation.
Discussion
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Figure 3. Effect of AA on infarct volume in female and SHRs. AA
(75 mg/kg) was administrated 6 hours after ischemia onset in
female pMCAO (A) and SHR tMCAO (B) models. Infarct volume
was determined at 24 hours after permanent MCAO by TTC
staining. AA treatment significantly reduced infarct volume compared with vehicle groups in both female and SHR. Scale bar, 1
cm. All values are means⫾SEM and analyzed by 1-way analysis
of variance. *P⬍0.05, **P⬍0.01, ***P⬍0.001 versus vehicle
group. AA indicates asiatic acid; SHRs, spontaneously hypertensive rats; pMCAO, permanent middle cerebral artery occlusion;
tMCAO, transient middle cerebral artery occlusion; MCAO, middle cerebral artery occlusion; TTC, triphenyltetrazolium chloride.
MCAO (Figure 5A). AA treatment restored State III. The
RCR (State III respiration divided by State IV respiration) is
an index of how coupled the electron transport chain is to
adenosine 5⬘-triphosphate production. Figure 5B shows alterations in RCR values after ischemia injury in both the
ipsilateral (injured) hemisphere and contralateral (uninjured)
hemisphere. After MCAO, RCR was significantly attenuated
in the ischemic cortex sample by 69.8%. AA treatment
decreased the attenuation of RCR by 42.8%.
Influence of AA on MMPs
The MMPs are induced in the brain after ischemia and are
involved in disruption of the BBB among other deleterious
processes.13 To determine whether AA treatment could inhibit MMP activity in the brain after ischemia, the levels of
MMP-2 and MMP-9 protein in the brain were determined by
We have previously shown in a mouse model of pMCAO that
AA is neuroprotective, has antioxidant activity, and protects
BBB integrity after ischemia.7 The current study, using
rigorous blinding and randomization methodology, has established a dose–response relationship and confirmed efficacy in
4 other MCAO stroke models: pMCAO/tMCAO in male rats,
pMCAO in female rats, and tMCAO in SHRs. We also show
that AA has a long and clinically relevant therapeutic time
window. Furthermore, we have identified ischemia-activated
deleterious mechanisms that are favorably influenced by AA.
Several of our findings are particularly noteworthy. First,
AA exhibited significant neuroprotection, even when administered 12 hours after the onset of ischemia, which could
translate to a clinically long time window for treatment and
will allow the treatment of many more patients. Second,
because almost half of all strokes affect females and hypertension is the biggest risk factor for stroke, efficacy in female
rats and the SHR model strengthens the case for further
preclinical development. Third, our findings that AA improves both histological and functional outcomes 14 days
after ischemia suggest sustained benefit.
Our pharmacokinetic data suggest that AA has a half-life
of 2.0 hours. This relatively short half-life raises the possibility that maintaining serum levels of AA for longer periods
of time by using multiple dosing or bolus plus infusion
regimens may afford greater efficacy. Future studies will
explore multiple dosing strategies to determine if there is
greater efficacy using those regimens. It is also possible that
multiple dosing regimens may allow smaller doses to be
administered.
AA favorably influences a number of deleterious mechanisms that are activated after stroke.2 We have previously
shown that AA is a powerful antioxidant and also that it
maintains the integrity of the BBB after ischemia.7 We have
now extended our mechanistic studies to include the influence of AA on mitochondrial redox activity and MMP
induction and enzymatic activity and show that AA also
favorably influences these pathways. The postischemic brain
exhibits prominent changes in redox activity of mitochondrial
respiratory chain components.14 –16 Mitochondria isolated
from ischemic brains exhibited decreases in State III respiratory rates of approximately 70% with nicotinamide-adenine
Lee et al
AA Protects Against Focal Cerebral Ischemia
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Figure 4. Improvement of long-term histological
and neurological functional outcomes after tMCAO.
AA (75 mg/kg) or vehicle were administered at 6
hours after ischemia onset. Infarct volume was
determined at 14 days after 2 hours tMCAO by
Nissl staining (A). AA treatment significantly
reduced infarct volume and improved functional
outcome. B, Neurological scores. C, Rotorod test.
D, Adhesive tape removal test at the right. AA
decreased motor, sensory, and reflex functional
damage after 2 hours tMCAO determined at 1, 3,
7, and 14 days after ischemia by neurological deficit scoring (0 –18). The adhesive removal and
Rotorod tests were done both before (baseline)
and 1, 3, 7, and 14 days after ischemia. There was
a strong trend toward better performance of the
AA-treated animals (white circle) in both Rotorod
and adhesive tape removal tests. Scale bar, 1 cm.
All values are means⫾SEM and analyzed by 1-way
analysis of variance. *P⬍0.05, **P⬍0.01,
***P⬍0.001 versus vehicle group. tMCAO indicates
transient middle cerebral artery occlusion; AA, asiatic acid.
dinucleotide-linked respiratory substrates.17 In the current
study, we confirm previous studies that showed that reperfusion after cerebral ischemia resulted in mitochondrial damage.18 –20 Our data show that AA inhibited ischemia induced
reductions of RCR, the index of mitochondrial respiratory
function indicating that AA protects mitochondria during
ischemia.
MMPs degrade the extracellular matrix and are involved in
several brain diseases including stroke.21,22 Enhanced MMP-9
expression and activity have been demonstrated in the ischemic brain.23,24 MMP inhibitors and MMP-9 gene deletion
attenuated BBB disruption and brain tissue infarction after
ischemia.25,26 Clinical studies have also shown a correlation
between plasma MMP-9 levels and the rate of hemorrhagic
transformation in human stroke.27 In the present study, we
confirmed that MMP-9 expression was increased after ischemia and showed that AA treatment attenuated the increase in
MMP-9. It is, however, possible that the attenuation in
Figure 5. Mitochondrial bioenergetics after tMCAO. A, Representative traces from nonsynaptic mitochondrial oxygen consumption measurements
in the presence of oxidative substrates (pyruvate and malate), ADP, oligomycin, CCCP, and succinate. B, The respiratory control ratios (RCR), which
is State III respiration divided by State IV respiration. Data were expressed as mean⫾SEM. *P⬍0.05, **P⬍0.01 versus contra group; #P⬍0.05,
##P⬍0.01 versus vehicle group. Contra indicates contralateral hemisphere; ipsil, ipsilateral hemisphere; tMCAO, transient middle cerebral artery
occlusion; ADP, adenosine 5⬘diphosphate; CCCP, carbonylcyanide m-chlorophenylhydrazone.
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Figure 6. Effects of AA on MMP-2 and
MMP-9. A, Western blot showing MMP-9
expression in the brain is enhanced after
ischemia in the ipsilateral hemisphere in
rats receiving vehicle (Lane 2), but this
effect is attenuated by AA (Lane 4).
MMP-2 levels were unchanged (␤-actin
illustrates protein loading in different lanes
for MMP-9 and MMP-2). B, MMP-2. C,
MMP-9. AA inhibited MMP-9 activity in
vitro regardless of excess ZnCl2. D,
MMP09. Data were expressed as
mean⫾SEM *P⬍0.05, **P⬍0.01 versus
contra group; #P⬍0.05, ##P⬍0.01,
###P⬍0.001 versus vehicle group. Contra
indicates contralateral hemisphere; ipsil,
ipsilateral hemisphere; AA, asiatic acid;
MMP, matrix metalloproteinase.
MMP-9 induction may be due to the smaller infarct volume
rather than the direct affect of AA on MMP induction. Our in
vitro data on the other hand show that AA inhibits MMP
enzyme activity. The mechanism through which AA inhibits
MMPs is not known; our study suggests that this inhibition is
not mediated by AA chelating zinc, an important cofactor for
MMPs. Inhibition of MMP-9 induction and/or activity may
underlie the maintenance of BBB integrity that we previously
reported with AA after focal ischemia.7
Many promising preclinical drugs have failed in clinical
testing. Reasons for failure include inadequate preclinical
testing and new guidelines (STAIR)8 have been published to
address this issue. Although we present considerable data
showing the efficacy, safety, and pharmacokinetics of AA,
more studies are still needed to fully meet the STAIR
guidelines and before AA can be tested in humans. Future
studies will test multiple dosing regimens including bolus
with infusion, compatibility with tissue-type plasminogen
activator, and efficacy in other comorbidity models and aged
animals. Pre-Investigational New Drug Food and Drug Administration mandated safety and toxicity studies would also
be needed.
In conclusion, we have shown that AA is neuroprotective
in multiple stroke models, it has a long therapeutic time
window, and favorably influences both early and late histological and functional outcomes. We also provide evidence
that AA protects mitochondria and attenuates MMP-9 induction and activity. Taken together these data suggest that AA
is an excellent candidate for further development as a stroke
therapy.
Sources of Funding
This work was supported by a National Institutes of Health R21 grant
to A.M. and the National Research Foundation of Korea Grant
funded by the Korean Government (NRF-2009 –352-E00061) to
K.Y.L.
Disclosures
None.
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Asiatic Acid Attenuates Infarct Volume, Mitochondrial Dysfunction, and Matrix
Metalloproteinase-9 Induction After Focal Cerebral Ischemia
Ki Yong Lee, Ok-Nam Bae, Kelsey Serfozo, Siamk Hejabian, Ahmad Moussa, Mathew Reeves,
Wilson Rumbeiha, Scott D. Fitzgerald, Gary Stein, Seung-Hoon Baek, John Goudreau, Mounzer
Kassab and Arshad Majid
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