Amelioration of learning and memory deficits by willughbeia cochinchinensis in mice

Nội dung text: Amelioration of learning and memory deficits by willughbeia cochinchinensis in mice

1. Journal of military pharmaco-medicine N o7-2017 AMELIORATION OF LEARNING AND MEMORY DEFICITS BY WILLUGHBEIA COCHINCHINENSIS IN MICE Nguyen Thi Hoa*; Le Van Quan*; Can Van Mao* SUMMARY Objectives: To study effects of willughbela cochinchinensis (WC) on learning and memory deficits in experimental animals. Subjects and methods: 50 Swiss mice were randomly separated into 5 experimental groups, 10 mice for each group. Group 1: mice were intraperitoneally (i.p) injected and orally (p.o) administered saline at dose 0.1 mL/10 g; group 2: mice were injected i.p 1.5 mg/kg scopolamin and p.o 0.1 mL/10 g saline; group 3, group 4 and group 5: mice were injected i.p 1.5 mg/kg scopolamin and p.o 100 mg/kg, 150 mg/kg and 200 mg/kg WC, respectively. 60 minutes after drug injections, animals performed a passive avoidance test which includes two phases: training phase: animals were placed in the light compartment and if they moved to the dark compartment, they were given electrical foot shocks for 3 seconds; test phase: animals were also placed in the light compartment but they were not given any electrical foot shocks at the dark compartment. Results: In test phase, mean latency to entry dark compartment in group 2 was shorter than that in group 1 and the latencies in group 4 and 5 were longer than that in group 2. Conclusion: Our results provided an evidence for effective treatment of WC in memory deficits animal model. * Keyword: Willughbela cochinchinensis; Learning and memory deficits; Mice. INTRODUCTION Up to date, there is no effective special treatments for AD and patients with AD Alzheimer’s disease (AD) accounts for must receive treatments in all their life. 60 - 80% of cases of dementia in older Furthermore, in some cases, effects of people [1]. Mechanism of AD has been suggested to be involved in drugs on AD treatments are limited. Thus, neurodegeneration and formations of development of new drugs and natural plaques and neurofibrillary tangles in plants for effective treatments of AD are brains which cause atrophied cortex and necessary. enlarged ventricles [2]. Following these It has indicated that dementia in patients damages in brains, patients with AD with AD relates to disorders in cholinergic develop deficits in memory, recognition systems. Thus, they used scopolamin, and behavioral controlling [3]. If they don’t cholinergic receptor antagonist, to induce receive any treatments, these disorders a animal model of AD. WC or “Gui do” will be worse and seriously affect to their has been used in Vietnamese traditional life as well as their families. medicines used for treatment of dementia * Vietnam Military Medical University Corresponding author: Can Van Mao (canvanmao2011@gmail.com) Date received: 10/06/2017 Date accepted: 10/08/2017 42
2. Journal of military pharmaco-medicine n o7-2017 as well as diarrhea, heartburn, and 2. Methods. subcutaneous abscess and as a diuretic. * Animal grouping and drug treatments: Our preliminary screening study revealed Animals were separated randomly into that methanolic extracts derived from the 5 experimental groups, 10 mice for each wood of W. cochinchinensis exhibit AChE group. Group 1 (control group): mice were (acetylcholinesterase) and BChE ip and p.o treated saline; group 2 (butyrylcholinesterase) strong inhibited (scopolamin group): mice were i.p treated activities which are main mechanism of scopolamin 1.5 mg/kg and p.o treated actions of drugs for treatments of AD [4]. saline at 0.1 mL/10 g; group 3, group 4 To provide basics for using WC to treat and group 5 (WC groups): mice were i.p AD in humans, we conducted the present injected scopolamin 1.5 mg/kg and p.o study with the aims: To investigate effects WC 100 mg/kg, 150 mg/kg and 200 of WC on deficits in learning and memory mg/kg, respectively. WC and saline were in experimental animals. orally administered at 60 minutes and scopolamin and saline were i.p injected at SUBJECTS AND METHODS 30 minutes before the behavioral task. 1. Subjects. * Passive avoidance test: 50 Swiss mice (150 - 250 g body Animals were required to perform weight) were used in the present study. passive avoidance test, which includes Animals were housed in individual cages, two phases: maintained in controlled temperature and - Training phase: was conducted at 12h light/dark cycles with free access to 60 minute after WC treatments on the first water and food. The present study was day. Passive avoidance box (Ugo Basile) conducted at Department of Physiology, was a chamber which contained Vietnam Military Medical University. All 2 compartments: light one and dark one. procedures were performed in There was a wall with a door to separate accordance with the Animal Center these compartments ( fig.1). The mice Guidelines for the Care and Use of were placed in light compartment and explore freely for 30 minutes. Then, the Laboratory Animals at the Vietnam door was raised to allow the mice to enter Military Medical University. the dark compartment. When the mice * Materials: entered the dark compartment, the door WC was isolated by Department of was closed and an electronic foot shock Pharmacy, Hochiminh City University of was delivered for 3 seconds. If mice didn’t Medicine and Pharmacy and was entered the dark compartment within supplied in power form. WC power was 300 seconds, mice were captured and dissolved in saline using a magnetic placed inside the dark compartment and a stirrer. foot shock was delivered for 3 seconds. 43
3. Journal of military pharmaco-medicine N o7-2017 Figure 1: Apparatus of passive avoidance test. - Test phase: were conducted on the - Latencies from beginning of trials to second day. Mice were placed in the light entrance into the dark compartment. Units compartment and the door was raised. In were measured as seconds (s). In the the test phase, when mice entered the present study, entry latencies were analyzed dark compartment, no foot shock was in the training phase and the test phase. delivered. Mice’s behaviors were recorded - Average speeds, units were measures for 300 seconds by using a digital video as meter/second (m/s). system. If animals didn’t enter the dark * Data analyses: compartment, entry latencies were Entry latencies and average speeds measured as 300 seconds and the trials were analyzed by one-way analysis of were over. variance (ANOVA) followed by the Tukey’s post-hoc test for multiple comparisons, * Research indicators: using SPSS 19.0. Results were considered In the present study, we analyzed to be statistically significant at p < 0.05. some research indicators, follow as: All results were expressed as mean ± SEM. RESULTS 1. Differences in entry latencies in the training phase. Figure 2: Entry latencies in training phase. 44
4. Journal of military pharmaco-medicine n o7-2017 Figure 2 showed mean entry latencies of experimental groups in the training phase. One way ANOVA indicated that there were no significant differences in entry latencies between experimental groups in the training phase (p > 0.05). 2. Differences in entry latencies in the test phase. Figure 3: Entry latencies in test phase. Figure 3 showed differences in entry latencies between experimental groups in the test phase. One way ANOVA indicated there was a significant main effects of experimental group [F(4.49) = 4.949; p = 0.002]. Post hoc test indicated that mean entry latency in the scopolamin group was significantly shorter than this in the control group (Tukey test, p < 0.05). Contrarily, entry latencies in WC 150 mg/kg WC 200 mg/kg treated groups were significantly longer than that in the scopolamin treated group (p < 0.001). 3. Differences in average speeds. Figure 4: Average speeds in experimental groups. 45
5. Journal of military pharmaco-medicine N o7-2017 Figure 4 showed differences in average than these of saline treated mice. These speeds between experimental groups. results are consisted with previous studies One way ANOVA indicated that there was [6, 7]. Interestingly, in the present study, a significant difference in average speeds we found that after WC treatments, there between experimental groups [F(4.49) were a significant increases in entry = 2.991, p = 0.029]. Post hoc test latencies of animals treated by WC at indicated that mean average speed was doses 150 mg/kg and 200 mg/kg, in significantly higher than that in the control compared to that of animals treated by group (Tukey test, p < 0.05). After WC scopolamin. These results indicated that treatments, mean average speeds in the WC ameliorated scopolamin-induced WC 150 mg/kg group and WC 200 mg/kg disorders in learning and memory in group were significantly lower than that in experimental animals. the scopolamin group (p < 0.05). Furthermore, in the present study, we also found that scopolamin induced DISCUSSION hyperactivities in experimental animals. Passive avoidance test is used widely These results are consistent with effects to evaluate learning and memory in of scopolamin to inhibit acetylcholinesterase rodents [5] Thus, this behavioral test is enzyme [8] and also with hyperactivities appropriate for the aim of our study. In the of patients with AD caused by disorders in present study, in the training phase, mice activities of the cholinergic system [9]. had no experience with dangerous events Interestingly, WC treatments also reduced (foot shocks) in the dark compartment. locomotordisorders of experimental animals. Thus, mice had tendencies to move to the The present’s results provided a dark compartment because of their important base for us to conduct next nocturnal life. These reasons induced, steps to apply WC for treatments of there was no significant difference in entry diseases relating to deficits In learning latencies between experimental groups. and memory, such as Alzheimer’s disease. In the test phase, learning and memory abilities of animals were expressed. When CONCLUSION animals had these better abilities, they In the present study, we demonstrated would recognize that when they moved to that WC reduced disorders in learning the dark compartment, they were received and memory as well as locomotion in foot shocks. Thus, the longer entry latencies experimental animals: they present, the better learning and - In the test phase of passive memory abilities of animals they have. In avoidance test, WC at doses 150 mg/kg the present study, scopolamin induced and 200 mg/kg increased entry latencies deficits in learning and memory abilities of in animals with scopolamin-induced animals expressed by differences in entry deficits in learning and memory. latencies between the control group and - WC at the same doses decreased scopolamin group. Entry latencies of average speeds of animals with scopolamin- scopolamin treated mice were shorter induced hyperactivities. 46
6. Journal of military pharmaco-medicine n o7-2017 ACKNOWLEDGEMENTS 5. Gacar N, Mutlu O, Utkan T, Komsuoglu C.I, Gocmez S.S, Ulak G. Beneficial effects of This work was supported by Grant resveratrol on scopolamine but not 106-YS.05-2013.24 from Vietnam’s mecamylamine induced memory impairment National Foundation for Science and in the passive avoidance and Morris water Technology Development (NAFOSTED). maze tests in rats. Pharmacol Biochem Behav. 2011, 99, pp.316-323. REFERENCE 6. Rush D.K. Scopolamine amnesia of 1. Alzheimer's Association . Alzheimer's passive avoidance: a deficit of information disease facts and figures. Alzheimers Dement. acquisition. Behav Neural Biol. 1988, 50 (3), 2014, 10 (2), e47-92. pp.255-274. 2. Madsen S.K, Gutman B.A, Joshi S.H, 7. Tabrizian K, Yaghoobi N.S, Iranshahi M, Shahraki J, Rezaee R, Hashemzaei M. Toga A.W, Jack C.R Jr, Weiner M.W, Auraptene consolidates memory, reverses Thompson P.M. Mapping dynamic changes in scopolamine-disrupted memory in passive ventricular volume onto baseline cortical avoidance task, and ameliorates retention surfaces in normal aging, MCI, and deficits in mice. Iran J Basic Med Sci. 2015, Alzheimer's disease. Multimodal Brain Image 18 (10), pp.1014-1019. Anal. 2013, 8159, pp.84-94. 8. Shannon H.E, Peters S.C. A comparison 3. Desai A.K, Grossberg G.T. Recognition of the effects of cholinergic and dopaminergic and management of behavioral disturbances agents on scopolamine-induced hyperactivity in dementia. Prim Care Companion J Clin in mice. J Pharmacol Exp Ther. 1990, 255 (2), Psychiatry. 2001, 3 (3), pp.93-109. pp.549-553. 4. Desmarais J.E, Gauthier S. Alzheimer 9. Khachiyants N, Trinkle D, Son S.J, Kim disease: clinical use of cholinergic drugs in K.Y. Sundown syndrome in persons with Alzheimer disease. Nat Rev Neurol. 2010, 6 dementia: an update. Psychiatry Investig. (8), pp.418-420. 2011, 8 (4), pp.275-287. 47