Name: experimental models to study the neuroprotection of acidic postconditioning against cerebral ischemia
Uploaded: 2017-07-31T12:30:00
Description: Watch this Scientific Journal Video about Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia at JoVE.com

This work was funded by the National Natural Science Foundation of China (81573406, 81373393, 81273506, 81221003, 81473186 and 81402907), Zhejiang Provincial Natural Science Foundation (LR15H310001) and the Program for Zhejiang Leading Team of S&amp;T Innovation Team (2011R50014).

All experiments were approved by and conducted in accordance with the ethical guidelines of the Zhejiang University Animal Experimentation Committee and were in complete compliance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Efforts were made to minimize any pain or discomfort, and the minimum number of animals was used.
1. OGD of Corticostriatal Slices
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	<li>Solution preparation:
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		<li>Prepare 1,000 mL r-ACSF (124 mmol/...

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Here we present two experimental models to study the neuroprotection of APC against cerebral ischemia. In brain slices, APC is achieved by incubating mice corticostriatal slices in acidic buffer bubbled with 20% CO2 after reperfusion onset, while in the MCAO model, APC is achieved by inhaling 20% CO2 to mice after reperfusion. Both models reflect the neuroprotection of APC against cerebral ischemia. The protection was comparable with that achieved by ischemic postconditioning but with a wider time w...

Stroke is one of the leading causes of mortality and disability worldwide. Great efforts have been made to find effective treatments for stroke in the last decades, however, the achievement is quite unsatisfactory. Postconditioning is a process manipulated by subtoxic stresses following an ischemic episode. Postconditioning, including ischemic, hypoxic, low-glucose and remote ischemic postconditioning, trigger endogenous adaptive mechanisms, and have been proven to be promising therapies against cerebral ischemia1,2,3,4. However, ischemic postconditioning may introduce additional injury. Limb remote ischemic postconditioning usually needs several cycles of 5 - 20 min occlusion and reperfusion on the ipsilateral or bilateral hind limbs5,6,7. Therefore, these postconditioning manipulations are dangerous or impractical in clinical practice. To overcome these disadvantages, we have developed APC as a therapy for focal cerebral ischemia in mice8. Induced simply by inhaling 20% CO2, APC significantly reduces ischemic brain injury in a more feasible and safer way. Recently we have proved that APC extends the reperfusion window, highlighting the significance of APC for stroke therapy9.
Here we present two experimental models to study the neuroprotection of APC against cerebral ischemia. The first one is the oxygen-glucose deprivation (OGD) model in mice corticostriatal slices. Rapid preparation and transfer of the brain slices into an artificial environment, usually artificial cerebrospinal fluid (ASCF), can maintain cell viability and neuronal circuitry, which makes it possible to study brain function in vitro10,11. OGD in ASCF mimics cerebral ischemia and induces ischemic injury12,13,14. After OGD, the brain slices are refreshed in regular ASCF (r-ASCF) to provide reperfusion and then treated with APC using acidic ASCF bubbled with 20% CO2. The corticostriatal slice maintains the intact histological characterization compared with primary cultured cells.
To study brain function in vivo, the mouse middle cerebral artery occlusion (MCAO) model is employed. The middle cerebral artery is blocked by inserting a flame-blunted monofilament via the common carotid artery. As one of the most widely used stroke models, the MCAO model shows clinical relevance and the application of a monofilament makes it easier to achieve reperfusion. Simply by inhaling normoxic mixed gas containing 20% CO2 after the onset of reperfusion, APC showed significant protective effects against cerebral ischemia indicated by reduced brain infarct volumes.

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experimental models to study the neuroprotection of acidic postconditioning against cerebral ischemia

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for neuroprotection, postconditioning treatments have proven to be promising therapies against cerebral ischemia. However, complicated procedures and potential safety issues limit their clinical application. To overcome these disadvantages, we have developed acidic postconditioning (APC) as a therapy for experimental focal cerebral ischemia. APC refers to the mild acidosis treatment by inhaling CO2 during reperfusion following ischemia. Here we present two models to execute APC in vitro and in vivo, respectively. The oxygen-glucose deprivation (OGD) treatment of mice and the corticostriatal occlusion and middle cerebral artery occlusion (MCAO) of mice were employed to mimic cerebral ischemia. APC can be simply achieved by transferring brain slices to acidic buffer bubbled with 20% CO2, or by mice inhaling 20% CO2. APC showed significant protective effects against cerebral ischemia, as reflected by tissue viability and brain infarct volume.

In the corticostriatal slice model described above, corticostriatal slice viability was quantified by TTC assay at 1 h after reperfusion. TTC conversion was calculated by normalizing the absorption at 490 nm to the control slice. According to TTC conversion, APC protected against OGD-induced reperfusion injury in an onset time and duration-dependent manner. In detail, both 1 and 3 min of acidosis treatment significantly improved viability at 5 min after 15 min OGD, whereas 5 min did not (...

Watch this Scientific Journal Video about Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia at JoVE.com

Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia

Acidic postconditioning protects against cerebral ischemia. Here we present two models to execute APC. They are achieved respectively by transferring corticostriatal slices to acidic buffer after oxygen-glucose deprivation in vitro and by inhaling 20% CO2 after middle cerebral artery occlusion in vivo.

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for ...

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