Korean J Physiol Pharmacol.  2016 May;20(3):279-286. 10.4196/kjpp.2016.20.3.279.

Neuroprotective effect of caffeic acid phenethyl ester in 3-nitropropionic acid-induced striatal neurotoxicity

• Bak J
• Kim HJ
• Kim SY
• Choi YS

• KMID: 2285600
• DOI: http://doi.org/10.4196/kjpp.2016.20.3.279

Abstract

Caffeic acid phenethyl ester (CAPE), derived from honeybee hives, is a bioactive compound with strong antioxidant activity. This study was designed to test the neuroprotective effect of CAPE in 3-nitropropionic acid (3NP)-induced striatal neurotoxicity, a chemical model of Huntington's disease (HD). Initially, to test CAPE's antioxidant activity, a 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) antioxidant assay was employed, and CAPE showed a strong direct radical-scavenging eff ect. In addition, CAPE provided protection from 3NP-induced neuronal cell death in cultured striatal neurons. Based on these observations, the in vivo therapeutic potential of CAPE in 3NP-induced HD was tested. For this purpose, male C57BL/6 mice were repeatedly given 3NP to induce HD-like pathogenesis, and 30 mg/kg of CAPE or vehicle (5% dimethyl sulfoxide and 95% peanut oil) was administered daily. CAPE did not cause changes in body weight, but it reduced mortality by 29%. In addition, compared to the vehicle-treated group, robustly reduced striatal damage was observed in the CAPE-treated animals, and the 3NP-induced behavioral defi cits on the rotarod test were signifi cantly rescued after the CAPE treatment. Furthermore, immunohistochemical data showed that immunoreactivity to glial fibrillary acidic protein (GFAP) and CD45, markers for astrocyte and microglia activation, respectively, were strikingly reduced. Combined, these data unequivocally indicate that CAPE has a strong antioxidant eff ect and can be used as a potential therapeutic agent against HD.

Keyword

3-nitropropionic acid; Antioxidant; Caff eic acid phenethyl ester; Huntington's disease; Striatum

MeSH Terms

Animals
Astrocytes
Body Weight
Cell Death
Dimethyl Sulfoxide
Glial Fibrillary Acidic Protein
Humans
Huntington Disease
Male
Mice
Microglia
Models, Chemical
Mortality
Neurons
Neuroprotective Agents*
Rotarod Performance Test
Urticaria
Dimethyl Sulfoxide
Glial Fibrillary Acidic Protein
Neuroprotective Agents

Figure

• 

  Fig. 1 Neuroprotective effect of CAPE in cultured striatal neurons.(A) The radical-scavenging activity of CAPE was measured with the ABTS antioxidant assay (n=4 for each group). (B) The neuroprotective effect of CAPE against 3NP intoxication was tested. For this study, 3NP and CAPE were treated and an LDH-release assay was performed two days later (n=4 for each group). Data are presented as mean±SEM, and significance was assessed using one-way analysis of variance (ANOVA). Post-hoc tests were performed using the Bonferroni correction. **p<0.01.

• 

  Fig. 2 Body-weight change and survival rates after 3NP injection in mice.(A) Body weight was measured at 12-h intervals during the 3NP injections (60, 60, 80, and 80 mg/kg) and daily for the remaining three days. There was no significant difference between the two groups. Data are presented as mean±SEM, and significance was assessed using Student's t-test. (B) The survival rate was 60% in the vehicle-treated group (6 out of 10 mice survived) and 89% in the CAPE-treated group (8 out of 9 mice survived).

• 

  Fig. 3 Rotarod analysis of 3NP-evoked fall latency.Motor activity was tested three times per day on an accelerating rotarod apparatus (4~40 rpm for 3 min), and the longest latency to fall off the rotarod during the three daily trials was used for analysis. The data from the surviving animals until three days after the last 3NP injection were included (n=6 and n=8 for the vehicle- and CAPE-treated groups, respectively). The data are presented as mean±SEM, and significance was assessed using Student's t-test. *p<0.05.

• 

  Fig. 4 CAPE reduced 3NP-evoked striatal damage.(A) The mice were perfused three days after the last 3NP injection and striatal damage was assessed with Cresyl Violet staining. The representative image shows distinct striatal damage in a vehicle-treated animal. The scale bar indicates 400 µm. (B) Quantitative analysis of striatal damage in the vehicle- and CAPE-treated groups. Quantitative measurements of striatal damage were analyzed with the χ2 test. *p<0.05.

• 

  Fig. 5 Astrocyte activation in the striatum.Representative images of GFAP immunoreactivity were shown from vehicle-treated animals (A and a1) and CAPE-treated animals (B and b1). Animals were perfused at 3 days after the last 3NP injection. Size bars are 400 and 50 µm in B and b1, respectively.

• 

  Fig. 6 Microglia/macrophage activation in the striatum.Representative images of immunoreactivity to CD45, a marker of activated microglia/macrophages, were shown from vehicle-treated animals (A and a1) and CAPE-treated animals (B and b1). Animals were perfused at 3 days after the last 3NP injection. Size bars are 400 and 50 µm in B and b1, respectively.

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