Korean J Physiol Pharmacol.  2017 Jan;21(1):55-64. 10.4196/kjpp.2017.21.1.55.

Dehydroevodiamine·HCl enhances cognitive function in memory-impaired rat models

• Shin KY
• Kim KY
• Suh YH

• KMID: 2371068
• DOI: http://doi.org/10.4196/kjpp.2017.21.1.55

Abstract

Progressive memory impairment such as that associated with depression, stroke, and Alzheimer's disease (AD) can interfere with daily life. In particular, AD, which is a progressive neurodegenerative disorder, prominently features a memory and learning impairment that is related to changes in acetylcholine and abnormal β-amyloid (Aβ) deposition in the brain. In the present study, we investigated the effects of dehydroevodiamine·HCl (DHED) on cognitive improvement and the related mechanism in memory-impaired rat models, namely, a scopolamine-induced amnesia model and a Aβ₁₋₄₂-infused model. The cognitive effects of DHED were measured using a water maze test and a passive avoidance test in the memory-impaired rat models. The results demonstrate that DHED (10 mg/kg, p.o.) and Donepezil (1 mg/kg, p.o.) ameliorated the spatial memory impairment in the scopolamine-induced amnestic rats. Moreover, DHED significantly improved learning and memory in the Aβ₁₋₄₂-infused rat model. Furthermore, the mechanism of these behavioral effects of DHED was investigated using a cell viability assay, reactive oxygen species (ROS) measurement, and intracellular calcium measurement in primary cortical neurons. DHED reduced neurotoxicity and the production of Aβ-induced ROS in primary cortical neurons. In addition, similar to the effect of MK801, DHED decreased intracellular calcium levels in primary cortical neurons. Our results suggest that DHED has strong protective effects against cognitive impairments through its antioxidant activity and inhibition of neurotoxicity and intracellular calcium. Thus, DHED may be an important therapeutic agent for memory-impaired symptoms.

Keyword

Alzheimer's disease; β-Amyloid; Cognitive function; Dehydroevodiamine; Scopolamine

MeSH Terms

Acetylcholine
Alzheimer Disease
Amnesia
Animals
Brain
Calcium
Cell Survival
Cognition Disorders
Cognition*
Depression
Dizocilpine Maleate
Learning
Memory
Models, Animal*
Neurodegenerative Diseases
Neurons
Rats*
Reactive Oxygen Species
Scopolamine Hydrobromide
Spatial Memory
Stroke
Water
Acetylcholine
Calcium
Dizocilpine Maleate
Reactive Oxygen Species
Scopolamine Hydrobromide
Water

Figure

• 

  Fig. 1 Effect of DHED or Donepezil on spatial memory impairment in scopolamine-injected rats.Training trials were carried out twice a day for 5 consecutive days. Scopolamine (1 mg/kg, i.p.) was given to rats immediately after the last training trial. DHED (10 mg/kg, p.o.) or Donepezil (1 mg/kg, p.o.) was given to rats 30 min after scopolamine administration. The trial tests were carried out at 1 h (1st), 4 h (2nd), and 24 h (3rd) after scopolamine administration. Latency is the time until a rat arrived at the platform. Note that at 1 h and 4 h the scopolamine-induced long latency was significantly decreased after DHED or Donepezil treatment. Data represents mean±standard error (SE). #p<0.05 compared with control and *p<0.05 compared with the scopolamine-injected group, one-way ANOVA.

• 

  Fig. 2 Effect of DHED on memory impairment in Aβ1-42-infused rats.(A) In Aβ1-42-infused rats, the training trial of the water maze test was performed after repeated administrations of DHED (10 mg/kg, p.o.) for 21 days. On trial day 5, latency was significantly increased in Aβ1-42-infused rats compared to the control group. However, latency was significantly decreased after DHED treatment compared to vehicle-treated Aβ1-42-infused rats. *p<0.05 compared to Aβ1-42-infused rats, one-way ANOVA. (B) The probe trial of the water maze test was carried out on day 2 after the final training trial. Note that the time spent in the platform quadrant (zone 1) was significantly decreased in the Aβ1-42-infused group compared to the control group. However, this time was significantly increased after DHED treatment compared to the Aβ1-42-infused only rats. Data represents mean±SE. #p<0.05 compared with Aβ1-42-infused rats and *p<0.05 compared with time in zone 1, one-way ANOVA. (C) In Aβ1-42-infused rats, which were generated via an infusion of the Aβ1-42 peptide into the lateral ventricle, a passive avoidance test was performed after repeated administrations of DHED (10 mg/kg, p.o.) for 21 days. Note that latency was decreased significantly in the Aβ1-42-infused group compared to the control group. However, latency was significantly increased after DHED treatment compared to the Aβ1-42-infused rats. Data represents mean±SE. *p<0.05 compared with the Aβ1-42-infused rats, one-way ANOVA.

• 

  Fig. 3 Effect of DHED on Aβ1-42-induced neurotoxicity and reactive oxygen species (ROS).(A) Treatment with Aβ1-42 peptide (25 µM) markedly induced neurotoxicity in primary neurons. However, the cell viability was significantly increased in a dose-dependent manner after pretreatment with DHED (0.5, 1, 2, and 4 µM) compared with the Aβ1-42-only-treated group (*p<0.05). In addition, cell viability was significantly increased after pretreatment with 50 µM of vitamin E compared with the Aβ1-42-only-treated group (*p<0.05). DHED or vitamin E was administered at 4 h before treatment with Aβ1-42. Data are expressed as a percent of the control value±SE. At least 2 experiments were carried out in triplicates. (B) ROS levels were significantly increased after treatment with the Aβ1-42 peptide. However, pretreatment with DHED (0.5, 1, 2, and 4 µM) significantly decreased ROS compared to the Aβ1-42-only-treated group (*p<0.05 and *p<0.01). In addition, pretreatment with 50 µM of vitamin E significantly decreased ROS compared to the Aβ1-42-only-treated group (*p<0.05). DHED or vitamin E was administered for 4 h before treatment with Aβ1-42. Data are expressed as a percent of the control value ± SE. At least 2 experiments were carried out in triplicates.

• 

  Fig. 4 Effect of DHED on intracellular Ca2+ levels in neurons.(A) Intracellular Ca2+ was assessed in neurons using confocal microscopy. Primary cortical neurons were treated with 1.2 mM of CaCl2 and 50 mM of KCl after 30 min of vehicle, 10 µM of MK801, or 10 µM of DHED treatment. Intracellular Ca2+ levels were measured after CaCl2 and KCl treatment. Note that intracellular Ca2+ levels were increased in vehicle-treated neurons. However, intracellular Ca2+ levels were decreased after pretreatment with MK801 or DHED compared to vehicle-treated neurons. DIC, differential interference contrast. (B) Quantification of intracellular calcium levels for analysis is shown. Data represents the mean±SE. *p<0.05 compared with vehicle-treated neurons, one-way ANOVA.

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