Experiments/Middle Cerebral Artery Occlusion (MCAO)

Source Paper

Cycloastragenol upregulates SIRT1 expression, attenuates apoptosis and suppresses neuroinflammation after brain ischemia

Man Li, Shi-chun Li, Bao-kai Dou, Ying-xiang Zou, Hao-zhen Han et al.

Acta Pharmacologica Sinica • 2020

DOI PMC

Middle Cerebral Artery Occlusion (MCAO)

behavioralmouseC57BL/6

Objective: Surgical induction of focal cerebral ischemia in mice via transient occlusion of the middle cerebral artery followed by reperfusion to test neuroprotective effects of CAG treatment

Materials & Equipment Checklist

6 items2 from ConductScience

Gather these items before starting the experiment. Check off items as you prepare.

Equipment1

Anesthesia delivery system for inhalational halothane

not specified • not specified • not specified • not mentioned

$2,099

Materials5

CAG (Caffeic acid phenethyl ester or similar compound)

Yuanye Biotechnology, Shanghai, China • not applicable • not specified • not mentioned

Amazon

DMSO (Dimethyl sulfoxide)

not specified • not applicable • not specified • not mentioned

Amazon

Tween 20 in PBS

not specified • not applicable • not specified • not mentioned

Amazon

Halothane

not specified • not applicable • not specified • not mentioned

Silicon-coated suture

not specified • not applicable • not specified • not mentioned

$400

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Estimated: $2,499.00

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Protocol Steps

Animal preparation and anesthesia

Male C57BL/6 mice (23-26g) were anesthetized with inhalational halothane at 1.5% concentration

not specifiednot specified

Small Animal Anesthesia Machine_TAIJI

Matches: Anesthesia delivery system for inhalational halothane

$2,099View

Note: Anesthesia was maintained during surgical procedure

View evidence from paper

“Male C57BL/6 mice (23–26 g, Vital River Laboratory Animal Technology Co., Ltd., Beijing, China) underwent right MCAO under inhalational anesthesia (1.5% halothane)”

Middle cerebral artery occlusion (MCAO)

Right MCAO was performed using a silicon-coated suture as previously described in references 15 and 16

45 minutesnot specified

Silicone Coated MCAO Monofilament Suture

Matches: Silicon-coated suture

$400View

Note: Occlusion was maintained for 45 minutes before reperfusion

View evidence from paper

“underwent right MCAO under inhalational anesthesia (1.5% halothane) as previously described. After 45 min, the silicon-coated suture was withdrawn”

Reperfusion initiation

The silicon-coated suture was withdrawn after 45 minutes of occlusion to initiate reperfusion

at 45 minutes post-occlusionnot specified

Silicone Coated MCAO Monofilament Suture

Matches: Silicon-coated suture

$400View

Note: This marks the beginning of the reperfusion phase

View evidence from paper

“After 45 min, the silicon-coated suture was withdrawn to initiate reperfusion”

CAG preparation

CAG (purity >98%) was dissolved in DMSO and then diluted with 2% Tween 20 in PBS to achieve a final DMSO concentration of 5%

not specifiednot specified

Note: CAG was prepared fresh for injection

View evidence from paper

“CAG (purity > 98%, Yuanye Biotechnology, Shanghai, China) was dissolved in DMSO and then diluted with 2% Tween 20 in PBS. The final concentration of DMSO was 5%”

Animal group assignment

Animals were randomly divided into three groups: sham, MCAO, and MCAO + CAG-treated groups

not specifiednot specified

Note: Randomization was performed to ensure unbiased group assignment

View evidence from paper

“The animals were randomly divided into the sham, MCAO, and MCAO + CAG-treated groups”

Initial CAG injection

CAG was injected intraperitoneally (i.p.) immediately after reperfusion at doses of 5, 10, or 20 mg/kg

immediately after reperfusionnot specified

Note: First injection administered right after suture withdrawal

View evidence from paper

“CAG (5, 10 or 20 mg/kg, i.p.) was injected immediately after reperfusion”

Second CAG injection

CAG was injected intraperitoneally at the same dose 12 hours after the initial injection

12 hours post-initial injectionnot specified

Note: Second dose administered at 12-hour interval

View evidence from paper

“CAG (5, 10 or 20 mg/kg, i.p.) was injected immediately after reperfusion, 12 h later”

Maintenance CAG injections

CAG was injected twice daily for up to three days following the initial injection

twice daily for three daysnot specified

Note: Continued dosing regimen to maintain therapeutic levels

View evidence from paper

“then twice daily for up to three days”