Lab Anim Res.  2017 Dec;33(4):308-314. 10.5625/lar.2017.33.4.308.

Quercetin attenuates the injury-induced reduction of γ-enolase expression in a middle cerebral artery occlusion animal model

• Jeon SJ
• Kim MO
• Ali-Shah F
• Koh PO

• KMID: 2402267
• DOI: http://doi.org/10.5625/lar.2017.33.4.308

Abstract

Quercetin, a natural flavonoid, copiously exists in vegetable, fruits and tea. Quercetin is beneficial to neurodegenerative disorders via its strong anti-oxidant and anti-inflammatory activities. γ-Enolase is one of the enzymes of glycolytic pathway and is predominantly expressed in neuronal cells. The aim of the present study is to verify whether quercetin modulates the expression of γ-enolase in brain ischemic injury. Adult Sprague-Dawley male rats were subjected to middle cerebral artery occlusion (MCAO) and quercetin (50 mg/kg) or vehicle was administered by intraperitoneal injection at 1 h before MCAO onset. A proteomics study, Western blot analysis, reversetranscription-PCR, and immunofluorescence staining were conducted to investigate the change of γ-enolase expression level. We identified a decline in γ-enolase expression in MCAO-operated animal model using a proteomic approach. However, quercetin treatment significantly attenuated this decline. These results were confirmed using Western blot analysis, reverse transcription-PCR, and immunofluorescence staining techniques. γ-Enolase is accepted as a neuron specific energy synthesis enzyme, and quercetin modulates γ-enolase in a MCAO animal model. Thus, our findings can suggest the possibility that quercetin regulates γ-enolase expression in response to cerebral ischemia, which likely contributes to the neuroprotective effect of quercetin.

Keyword

cerebral ischemia; neuroprotection; quercetin

MeSH Terms

Adult
Animals
Blotting, Western
Brain
Brain Ischemia
Fluorescent Antibody Technique
Fruit
Humans
Infarction, Middle Cerebral Artery*
Injections, Intraperitoneal
Male
Middle Cerebral Artery*
Models, Animal
Neurodegenerative Diseases
Neurons
Neuroprotection
Neuroprotective Agents
Proteomics
Quercetin*
Rats
Rats, Sprague-Dawley
Tea
Vegetables
Neuroprotective Agents
Quercetin
Tea

Figure

• 

  Figure 1 γ-Enolase protein spots identified by MALDI-TOF in the vehicle+sham, quercetin+sham, vehicle+middle cerebral artery occlusion (MCAO), and quercetin+MCAO animals. Squares indicate the γ-enolase protein spots (A). The intensity of spots was measured using PDQuest software (B). The ratio of intensity is described as spots intensity of these animals to spots intensity of sham+vehicle animals. Data (n=4) are shown as mean±SEM. *P<0.05.

• 

  Figure 2 Western blot analysis of γ-enolase protein levels in the vehicle+sham, quercetin+sham, vehicle+middle cerebral artery occlusion (MCAO), and quercetin+MCAO animals. Each lane represents an individual animal (A). Densitometric analysis is represented as intensity of γ-enolase to intensity of β-actin (B). Data (n=4) are shown as mean±SEM. *P<0.05.

• 

  Figure 3 Reverse transcription-PCR analysis of γ-enolase protein levels in the vehicle+sham, quercetin+sham, vehicle+middle cerebral artery occlusion (MCAO), and quercetin+MCAO animals. Each lane represents an individual animal (A). Densitometric analysis is represented as intensity of γ-enolase to intensity of β-actin (B). Data (n=4) are shown as mean±SEM. *P<0.05.

• 

  Figure 4 Images of double immunofluorescence labeling with γ-enolase (green color) and DAPI (nuclei marker, blue) in the ischemic core of cerebral cortex in vehicle+sham, quercetin+ sham, vehicle+middle cerebral artery occlusion (MCAO), and quercetin+MCAO animals (A). Quantitative assessment of γ-enolase positive neurons in ischemic core of rat cerebral cortex (B). The arrows indicate the γ-enolase positive cells. Data (n=4) are shown as mean±SEM.*P<0.05. Scale bars=100 µm.

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