Willughbeia cochinchinensis ameliorates locomotor disorders in model of alzheimer’s disease mice

Nội dung text: Willughbeia cochinchinensis ameliorates locomotor disorders in model of alzheimer’s disease mice

1. Journal of military pharmaco-medicine N o7-2017 WILLUGHBEIA COCHINCHINENSIS AMELIORATES LOCOMOTOR DISORDERS IN MODEL OF ALZHEIMER’S DISEASE MICE Can Van Mao*; Tran Hai Anh*; Le Van Quan* SUMMARY Objectives: To evaluate the effects of willughbela cochinchinessis (WC) to locomotor disorders in an Alzheimer's disease (AD) model of mice. Subjects and methods: 50 Swiss mice were separated randomly into 5 experimental groups, 10 mice for each group. Group 1: Mice were intraperitoneally injected (i.p) and orally administered (p.o) saline at dose 0.1 mL/10 g; group 2: mice were injected i.p scopolamin 1.5 mg/kg and p.o saline 0.1 mL/10 g; group 3, group 4 and group 5: mice were injected i.p scopolamine 1.5 mg/kg and p.o WC 100 mg/kg, 150 mg/kg and 200 mg/kg, respectively. WC and saline were orally administered at 60 minutes and scopolamin and saline were injected i.p at 30 minutes before the behavioral task. 60 minutes after WC injections, mice were placed in an open field for 5 minutes. Behaviors of mice were observed by a camera and analyzed by Anymaze software. Results: WC at doses 150 mg/kg and 200 mg/kg reversed scopolamin-induced hyperactivities in mice. Conclusion: These results provided a basic for developing a new drug to treat patients with AD. * Keywords: Alzheimer's disease; Willughbela cochinchinessis; Scopolamin; Locomotor behaviors; Mice. INTRODUCTION been demonstrated that animals also Alzheimer’s deasease is one form of exhibit increased locomotor activities dementia in older humans. Mechanism of including hyperactivity, stereotypic behaviors, this kind disease has been suggested to and home cage activity disturbances [4]. be involved in neurodegeneration and It has been suggested that disorders of formation of plaques and neurofibrillary neurotransmitter systems, especially tangles [1]. These changes in the brain cholinergic system and glutamate systems cause behavioral disorders such as are associated with abnormal behaviors cognitive and memory impairments and of patients with AD [5]. Furthermore, locomotor hyperactivities. In patients with abnormal activity of the cholinergic system AD, it has been shown that patients with affects glutamatergic systems [6]. Thus, AD expressed hyperactivities in the late it has developed a animal model of AD afternoon and the evening. These by injecting intraperitoneally scopolamin, hyperactivities are termed as sundown a form of anticholiergic drug and used syndrome or sundowing [2, 3]. In animal this model to study effects of new drugs models of AD, previous studies have or natural plants in animal models of AD. * Military Medical University Corresponding author: Cao Van Mao (caovanmao2011@gmail.com) Date received: 15/06/2017 Date accepted: 10/08/2017 16
2. Journal of military pharmaco-medicine n o7-2017 Recently, we applied this model to evaluate 1.5 mg/kg and p.o treated saline at 0.1 effects of WC to scopolamin-induced mL/10 g; group 3, group 4 and group 5 deficits in cognition and memory in mice. (WC groups): mice were i.p injected However, patients and scopolamin induced scopolamin 1.5 mg/kg and p.o WC 100 mg/kg, animals with AD also showed disorders in 150 mg/kg and 200 mg/kg, respectively. locomotion [2, 3, 7, 8]. Thus, effects of WC and saline were orally administered WC to locomotor disorders should be at 60 minutes and scopolamin and saline evaluated. We conducted the present study were i.p injected at 30 minutes before the with the aim: To investigate effects of WC behavioral task. to ameliorate locomotor disorders in an * Open field test: animal model of AD. 60 minutes after drug treatments, mice SUBJECTS AND METHODS were placed in the center of a open field box. Open field box was a square box 1. Subjects. (40 × 40 × 60 cm), covered with 50 Swiss mice (150 - 250 g body weight) polypropylene sheets inside the wooden were used in the present study. Animals box (figure 1). Animals were allowed to free were housed in individual cages, maintained explore inside open field box for 5 minutes. in controlled temperature and 12h light/dark Behaviors of animals were recorded using cycles with free access to water and food. a digital video system. Data was analyzed The present study was conducted at offline by ANY-maze software (Stoelting Department of Physiology, Vietnam Military Co., Wood Dale, IL, USA). Medical University. All procedures were performed in accordance with the Animal Center Guidelines for the Care and Use of Laboratory Animals at the Vietnam Military Medical University. 2. Materials. WC was isolated by Department of Pharmacy, Hochiminh City University of Medicine and Pharmacy and was supplied in power form. WC power was dissolved in saline using a magnetic stirrer. 3. Methods. * Animal grouping and drug treatments: Figure 1: Open field box. Animals were separated randomly into * Research indicators: 5 experimental groups, 10 mice for each group: group 1 (control group): mice were ip In the present study, we analyzed some and p.o treated saline; group 2 (scopolamin research indicators as followed: group): mice were i.p treated scopolamin - Travel distances (m). 17
3. Journal of military pharmaco-medicine N o7-2017 - Average speeds (m/s). analyzed by one-way analysis of variance - Ratios of mobile time/immobile time. (ANOVA) followed by the Tukey’s post- hoc test for multiple comparison, using * Data analysis: SPSS 19.0. Results were considered to Travel distance, travel speed and ratio be statistically significant at p < 0.05. All of mobile time/immobile time were results were expressed as mean ± SEM. RESULTS 1. Changes in travel distance. Figure 2: Travel distance of mice. Figure 2 showed travel distance of mice in the open field test. There was a significant difference in travel distance of mice in the experimental groups [F(5.49) = 3.82, p = 0.016]. Post hoc test indicated that mean travel distance of mice in the scopolamin group was significantly longer than this in the control group (Tukey test, p < 0.05). After WC treatment, travel distance of mice decreased gradually from WC 100 mg/kg to WC 200 mg/kg. However, a significant decrease in mean travel distance of mice was observed in only WC 200 mg/kg group (Tukey test, p < 0.05). 2. Changes in average speed. Figure 3: Average speed of mice. 18
4. Journal of military pharmaco-medicine n o7-2017 Figure 3 showed changes in average speed of mice in the open field test. One way ANOVA indicated that there was a significant difference in average speed of mice in experimental groups [F(4.49) = 4.12, p = 0.011]. Post hoc test indicated that there was a significant increase in average speed of mice in scopolamin group, compared to this in the control group (Tukey test, p < 0.05). After WC treatments, average speed also reduced gradually from WC 100 mg/kg to WC 200 mg/kg. However, compared to average speed of mice in control group, a significant decrease in average speed was also observed in the WC 200 mg/kg group only (Tukey’s test, p < 0.05). 3. Changes in ratio of mobile time/immobile time. Figure 4: Ratio of mobile time/immobile time of mice. Figure 4 showed changes in ratio of in the scopolamin group (Tukey test, p < mobile time/immobile time of mice in the 0.05). experimental groups. One way ANOVA Ratios of mobile time/immobile time of indicated that there was a significant mice expressed mobile or immobile difference in ratio of mobile time/immobile tendencies of animals. If this ratio is time of mice between experimental groups higher than 1, it indicates that animals [F(4.49) = 6.11, p = 0.01]. Post hoc test might tend to be mobile. On the contrary, indicated that mean ratio of mobile it indicates that animals might tend to be time/immobile time of mice in the immobile. Results in figure 4 indicated that scopolamin group was significantly higher WC at doses 150 mg/kg and 200 mg/kg than this in the control group (Tukey test, reversed scopolamin induced increasing p < 0.05). When mice were treated by WC, mobile tendencies of mice. these ratios decreased gradually. Ratios of mobile time/immobile time of mice in DISCUSSION WC 150 mg/kg group and WC 200 mg/kg Hyperactivity is one of behavioral group were significantly lower than those disorders in both patients with AD and 19
5. Journal of military pharmaco-medicine N o7-2017 animal models of AD. In patients with AD, CONCLUSION it was demonstrated that they display In the present study, we used open sleeping and locomotion disorders. These field test to investigate effects of WC on disorders are more serious in the late scopolamin induced locomotor disorders afternoon and evening. Thus, these disorders in animals with Alzheimer-like symptoms. were called as sundown syndrome or Our results indicated that WC at doses sundowning [2, 3]. Locomotor disorders 150 mg/kg and 200 mg/kg reduced are observed in animal models of AD, scoplamin-induced hyperactivities in including the scopolamin-induced Alzheimer experimental animals. The reduction of model [7, 8]. In these models, mice exhibit hyperactivities was expressed by significant hyperactivity tendencies during the dark decreases in travel distance, average phase (the active phase of mice) [4]. Thus, in developments of new drugs or speed and ratio of mobile time/immobile natural plants to treat for AD, study on timey. These results provided important locomotor functions of experimental animals basic for next researches to use WC for is necessary. treatment of AD in humans. Open field test used widely to investigate ACKNOWLEDGEMENTS locomotor functions in small animals such as rats and mice. This test allows to evaluate This work was supported by Grant 106- many research indicators. Thus, this test YS.05-2013.24 from Vietnam’s National will help us to study more particularly and Foundation for Science and Technology exactly the effects of new drugs to Development (NAFOSTED). locomotor functions of experimental animals [9]. In the present study, the open REFERENCE field test was used to assess effects of WC 1. Crews L, Masliah E . Molecular mechanisms to scopolamine- induced hyperactivities of of neurodegeneration in Alzheimer's disease. animals with Alzheimer-like symptoms. Hum Mol Genet. 2010, 19(R1):R12-20. Results showed changes in some 2. Vitiello M.V1, Bliwise D.L, Prinz P.N. research indicators to indicate that WC Sleep in Alzheimer's disease and the sundown ameliorated disorders in locomotor syndrome. Neurology. 1992, 42 (7 Suppl 6), functions of animals. These are: WC has pp.83-93. effects to decrease travel distance, 3. Bliwise D. L. What is sundowning?. J average speed and ratio of mobile Am Geriatr Soc. 1994, 42, pp.1009-1011. time/immobile time of animals with 4. Ambree O, Touma C, Gortz N, Keyvani scopolamin-induced hyperactivities. These K, Paulus W, Palme R et al. Activity changes results along with our previous study’s and marked stereotypic behavior precede results [6] provided a important basic to Abeta pathology in TgCRND8 Alzheimer apply new plants for treating AD in humans. mice. Neurobiol.Aging. 2006, 27, pp.955-964. 20
6. Journal of military pharmaco-medicine n o7-2017 5. Terry A.V Jr, Buccafusco J.J. The 7. Sanberg P.R, Henault M.A, Hagenmeyer- cholinergic hypothesis of age and Alzheimer's Houser S.H, Russell K.H. The topography disease-related cognitive deficits: recent of amphetamine and scopolamine-induced challenges and their implications for novel hyperactivity: toward an activity print. Behav drug development. J Pharmacol Exp Ther. Neurosci. 1997, 101 (1), pp.131-133. 2003, 306 (3), pp.821-827. 8. Furuie H, Yamada K, Ichitani Y. MK- 6. Van Can M, Tran A.H, Pham D.M, Dinh 801-induced and scopolamine-induced B.Q, Van Le Q, Van Nguyen B, Thi Nguyen hyperactivity in rats neonatally treated M.T, Nguyen H.X, Nguyen N.T, Nishijo H. chronically with MK-801. Behav Pharmacol. Willughbeia cochinchinensis prevents 2013, 24 (8), pp.678-683. scopolamine-induced deficits in memory, 9. Seibenhener M.L, Wooten M.C. Use of spatial learning, and object recognition in the open field maze to measure locomotor rodents. J Ethnopharmacol. 2017, doi: 10.1016/ and anxiety-like behavior in mice. J Vis Exp. j.jep.2017.06.035. 2015, 96:e52434. doi: 10.3791/52434. 21