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)....
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 effect. 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 deficits on the rotarod test were significantly 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 effect and can be used as a potential therapeutic agent against HD.
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 effect. 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 deficits on the rotarod test were significantly 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 effect and can be used as a potential therapeutic agent against HD.
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문제 정의
This study was designed to evaluate the therapeutic potential of CAPE in 3NP-induced striatal neurotoxicity in a chemical model of HD. In the in vitro experiment, CAPE showed antioxidant activity, and it significantly reduced the cultured striatal neuronal cell death caused by 3NP.
가설 설정
In this study, we investigated whether CAPE could be used as a therapeutic agent against HD. For this purpose, we employed a 3-nitropropionic acid (3NP)-induced striatal neurotoxicity model in mice.
제안 방법
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.
(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).
For this study, we tested 1, 20, and 50 μM of CAPE, and as shown in Fig. 1A, the radical-scavenging activity was dose-dependently increased, indicating that CAPE has strong radical-scavenging activity.
Repeated administration of 3NP is well known to induce striatal neurotoxicity and behavioral deficits reminiscent of HD [20], so it has been widely used to generate HD in rodents. In this study, CAPE(30 mg/kg i.p.) was administered to mice starting at 30 min before the first 3NP injection and then daily until three days after the last 3NP injection. As shown in Fig.
The mice were trained on an accelerating rotarod (RotaRod-M; Jeung Do Bio & Plant, Korea) with three trials per day for three consecutive days.
3NP induces behavioral deficits reminiscent of HD pathogenesis, so to test whether CAPE rescues behavioral deficits, we employed the rotarod test. This test is used to evaluate motor coordination by measuring the latency to fall from the rotarod [24]. To our surprise, daily administration of CAPE significantly rescued motor deficits on the rotarod.
To test the neuroprotective effects, 20 or 50 μM of CAPE was treated with 5 mM of 3NP for two days, and cytotoxicity was measured with the LDH release assay.
For each section, the total striatal area and the damaged area were outlined, and the areas were measured using PhotoshopⓇ software. Volumes were calculated by summing the crosssectional area of each section, and the percentage of striatal damage in each animal was calculated by dividing the damaged volume by the total striatal volume. Striatal damage was measured on the following scale: no striatal damage, grade 0; less than 10% striatal damage, grade 1; 10–20% striatal damage, grade 2; higher than 20% striatal damage, grade 3.
데이터처리
Data are presented as mean±SEM, and significance was assessed using Student’s t-test.
Data are presented as mean±SEM, and significance was assessed using one-way analysis of variance (ANOVA).
The data are presented as mean±SEM, and significance was assessed using Student’s t-test.
The data were presented as mean±SEM, and significance was assessed using Student’s t-test.
The data were presented as mean±SEM, and significance was assessed using a log-rank test for the survival results and a two-tailed Student’s t-test for the motor behavior results.
The data were presented as mean±SEM, and significance was assessed using one-way analysis of variance (ANOVA).
이론/모형
The data were presented as mean±SEM, and significance was assessed using one-way analysis of variance (ANOVA). Post-hoc tests were performed using the Bonferroni correction.
To measure the free-radical-scavenging activity of CAPE, the ABTS antioxidant assay was performed according to the manufacturer’s instructions (Zenbio, Research Triangle Park, NC, USA).
성능/효과
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).
후속연구
Therefore, although the possibility that expression of antioxidant proteins plays an important role in CAPE-mediated neuronal protection cannot be ruled out, it is postulated that the direct radical-scavenging activity of CAPE is very important in providing neuroprotection. Further research is needed to elucidate the exact neuroprotective mechanism of CAPE in 3NP-induced striatal neurotoxicity.
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