|Year : 2019 | Volume
| Issue : 1 | Page : 36-40
Neurobehavioral activity study of methanolic whole plants extract of Cyperus rotundus Linn.
Imonul Kabir1, Subir Biswas1, Md Asaduzzaman2, Md Islam Molla1, Md Rajdoula Rafe1
1 Department of Pharmacy, Jagannath University, Dhaka, Bangladesh
2 Department of Zoology, Jagannath University, Dhaka, Bangladesh
|Date of Web Publication||22-Aug-2019|
Md Rajdoula Rafe
Department of Pharmacy, Jagannath University, Dhaka 1100
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Cyperus rotundus commonly known as “nutgrass” is extensively used in traditional Chinese and Indian Ayurvedic medicine. It is traditionally used to treat fevers, digestive disorders, wound, and bruises. Materials and Methods: This study evaluated the sedative-hypnotic and antidepressant effect of the methanolic extract of C. rotundus (MECR). To perform this study, the whole plants of C. rotundus were taken for extraction with methanol following soaking process and tested for acute toxicity on mice first. The sedative and hypnotic activity were then studied performing hole board and open field tests in albino mice model at the doses of 100 and 200 mg/kg body weight of MECR. Diazepam at the dose of 1 mg/kg was utilized as a standard drug in both experiments. Similarly, antidepressant activity test was also performed using forced swimming test and tail suspension test. Nortriptyline was used as a standard to assess antidepressant activity. Results: We found that MECR produced an insignificant dose-dependent effects against locomotors activity of mice both in hole cross and open field tests. Besides, it was also noticed after analyzing forced swimming and tail suspension test that it has no significant antidepressant activity. Conclusion: Taken together, our study suggests that MECR do not possess notable sedative-hypnotic and antidepressant or neurobehavioral properties.
Keywords: Antidepressant, Cyperus rotundus, insignificant, sedative-hypnotic, toxicity
|How to cite this article:|
Kabir I, Biswas S, Asaduzzaman M, Molla MI, Rafe MR. Neurobehavioral activity study of methanolic whole plants extract of Cyperus rotundus Linn. J Pharm Negative Results 2019;10:36-40
|How to cite this URL:|
Kabir I, Biswas S, Asaduzzaman M, Molla MI, Rafe MR. Neurobehavioral activity study of methanolic whole plants extract of Cyperus rotundus Linn. J Pharm Negative Results [serial online] 2019 [cited 2019 Sep 23];10:36-40. Available from: http://www.pnrjournal.com/text.asp?2019/10/1/36/265141
Authors Imonul Kabir and Subir Biswas contributed equally
| Introduction|| |
Medicinal plants have eminent qualities for producing new drugs of massive advantages to the humankind. There are different ways to explore new biologically active mechanism of plants. Many secondary metabolites are generally produced from plants that comprise an important source for pharmaceutical products. The opportunity of using the plant as a source of medicine is huge due to the wide diversity and availability of plants around the world. Along with other activities, attempts have also been made to find out newer sedative-hypnotic drugs from different kinds of plants. Sedative and hypnotics are the drugs that can alleviate anxiety and generate a calming effect by initiating the commencement of sleep and preserving sleeping duration. At present, these drugs are broadly applied in the treatment of several psychiatric disorders which comprise insomnia and anxiety. Insomnia is termed as static inconvenience in falling or staying asleep which can influence important physical and psychological disorder. Sedatives are that types of drugs which diminish the action, inducing a relaxing and calming effect. At higher doses, sedatives generally produce sleep. Those drugs which are used predominantly to cause sleep are called hypnotics. The distinction among sedatives and hypnotic is generally the quantity of the dose; lesser doses undergo a serene effect and greater doses produce sleep. Incessant use of these currently obtainable sedative-hypnotic therapies inclined to have significant undesirable effects ranging from respiratory, digestive, physical dependence, and tolerance. The improvement of newly hypnotic-sedative drugs with lesser undesirable effects has been suggested to be a promising approach to combat different psychiatric disorders.
Cyperus rotundus Linn. (Nutgrass, family Cyperaceae) is extensively distributed in numerous tropical and subtropical territory of the world. In Bangladesh, C. rotundus is mostly known as “motha” or “bada.” The term Cyperus is derived from Cypeiros, which was the ancient Greek name for the genus, rotundus is Latin word for round and refers to the tuber. It is an erect, smooth, and perennial medicinal plant having scaled, wiry, creeping, slender, tenebrous, and persistent rhizomes. Phytochemical analysis has demonstrated that the principal chemical components of this herb are essential oils, terpenoids, mono sesquiterpenes, and flavonoids. The plant includes the following chemical components: acyperone, isocyperol, cyperotundone, mustakone, cyprotene, acopaene, cyperene, aselinene, rotundene, valencene, cyperol, gurjunene, 1,8-cineole and 4,11-selinnadien-3-one.
Tubers of C. rotundus are used to treat excess bleeding, loss of appetite, boils, blisters, diarrhea, cough, fevers, lacteal disorders, inflammation, rheumatoid arthritis, stomach ailments, skin rashes, vomiting, excessive thirst, wounds, and worm infestation.,, Furthermore, they are used as a remedy for renal colic and dysentery. Moreover, the plant possesses several biological properties such as antioxidant, cytotoxic,,, antiallergic , insecticides,, antimalarial, antimicrobial,, antidiarrheal, antipyretic, inflammatory, antiemetic, hypotensive  hepatoprotective,, antidiabetic, and anticonvulsant.
Here, we aim to explore new activities of C. rotundus which were not investigated earlier. Therefore, we tried to evaluate sedative-hypnotic and antidepressant activities of methanolic extract of whole plant to understand its neurobehavioral characteristics.
| Materials and Methods|| |
The plant collection and extraction
The whole plants of C. rotundus were collected from Brahmanbaria, Bangladesh, in January 2019. The samples were then identified by Mahbuba Sultana, Senior Scientific Officer (Deputation), Bangladesh National Herbarium, Mirpur, Dhaka, Bangladesh. An acknowledgement receipt (DACB: 46873) has been kept in the Herbarium for further reference. The whole plants were then washed thoroughly for removing dirty materials with episodic sun drying and shade-dried for several days. Then, these were dried in an oven at notably low temperature for 24 h for better crushing. The dried plants were pulverized into coarse powder by a crushing machine in the laboratory of Department of Pharmacy, Jagannath University. Then, powdered dried plants (400 g) were macerated with approximately 2 L of methanol with extemporaneous agitation at 25°C ± 2°C for 10 days in an amber color glass bottle. Then, a sterilized cotton filter and Whatman No. 1 filters paper were used to filter the extract. For further studies, this crude extract was used.
Healthy Swiss albino mice (male and female) of 25–30 g were collected from Animal Resources Branch of the International Center for Diarrhoeal Disease Research, Bangladesh (ICDDR, B). The animals were placed in standard laboratory environment (room temperature 25°C ± 2°C; relative humidity 55%–60%; 12 h light/dark cycle) and were supplied with adequate amount of standard diet (ICDDR, B formulated) and clean water during acclimatization period. Before conducting our experiments, the animals were adapted to the laboratory conditions for 15 days. Experimental mice were treated according to the moral principles and guidelines for scientific experiments on animals (1995) created by the Swiss Academy of Sciences and the Swiss Academy of Medical Sciences.
Standard drugs and plant samples were prepared on the day of the experiments to administer freshly. Mice of control group were treated with saline water (0.1 ml/mice) as vehicle. Diazepam (1 mg/kg body weight [BW], for sedative activity) and nortriptyline (1 mg/kg BW, for antidepressant activity) were used to compare as standard drugs.
Acute toxicity test
The mice were divided into control and three test groups each containing five animals. MECR was administered to the mice orally at doses of 500, 1000, and 2000 mg/kg. The mice were given access to sufficient amount of food and water, and all mice were noticed for mortality and allergic symptoms for the next 14 days.
Open field test
This test is broadly used for the evaluation of emotive behavior of the animals, particularly the rodents. This process was performed as described by Gupta et al. Open field apparatus was constructed by a wooded field of half square meter with an order of squares and drawn in white and black color. It had a 30 cm height wall and was placed in a low light condition. First, mice were divided into four groups by using black, green, blue, and red color permanent marker and containing 5 mice each for standard, control, and test samples. Then, saline water, two doses of extract (100 and 200 mg/kg), and diazepam were orally administered consecutively to the marked mice. Treated mice were placed in the center of the open field for 3 min for each at 30, 60, 90, and 120 min after the treatments. In every 3 min, the number of squares visited by the mice were counted and noted down.
This test is used to measure emotionality, stress, neophilia, and anxiety in animals. The hole-board test was accomplished with little modifications to previously narrated process of Oztürk et al. Hole-board test apparatus was constructed using a perforated wood floor board with 60 cm × 30 cm in diameter and 16 evenly spaced holes. Experimental mice were divided into four groups by using black, green, blue, and red color permanent marker and containing 4 mice each for standard, control, and test samples. Then, control solvent (saline water), 100 and 200 mg/kg test samples, and diazepam as standard sample were orally administered to the marked mice. After 45 min, each mouse was placed in the center of the hole board for 5 min consecutively and allowed to move on the board. For each animal, the number of head dips into the holes was counted and noted down for 5 min.
Forced swimming test
The forced swimming test (FST) has been used extensively to evaluate the antidepressant efficacy of new compounds that intended to preventing or rendering depressive-like condition. This method is based on observing the animals exposed to a condition of swimming forcefully that they become immovable and lethargic after a period of an energetic activity (struggling) and generating only the movements required to keep their heads above the water. The FST was completed on mice following the method published by Porsolt et al. For this test, an individual Plexiglas's cylinders with 40 cm high and 24 cm diameter were used and filled the cylinder up to 20 cm by water. First, mice were divided into four groups using four different color permanent marker and containing 5 mice to each group for standard, control, and test samples for FST. Then, test samples, control, and standard sample (nortriptyline) were orally administered to the marked mice. After 45 min of administering samples, swimming sessions were performed by putting the treated mice in individual Plexiglas's cylinders. All the mice were forced to swim for 6 min, and the period of time spent in immovability during the final 5 min of a 6 min observation period was recorded. Mice are considered as immobile when floating without movement or making only those movements necessary to keep the head above water.
Tail suspension test
The tail suspension test is broadly useful rodent behavioral experiment to evaluate the effect of depression-related behavior and potential antidepressant properties. The total duration of immovability induced by tail suspension was measured following to the process stated by Steru et al. One compartment rectangular tail suspension box was made using wood (60 cm in total length, 40–55 cm in height, 15 cm in width, and 11.5 cm in depth). For this experiment, mice were divided into four groups by using black, green, blue, and red color permanent marker and containing 5 mice each for standard, control, and test samples. Then, plant samples, control, and standard sample (nortriptyline as standard) were orally administered. After 45 min, mice were suspended middle of the compartment which was 35 cm on the floor by fixative tape placed about 2–3 cm from the tip of the tail. After that, immobility time was manually documented during a 5 min period of time consecutively for each mouse. Mice were considered immobile only when they overhung motionless or stayed completely passive.
All data were processed and analyzed using MS Excel version 2013. The results obtained from the tests were expressed as mean ± standard error of mean of five animals.
| Results|| |
The crude extracts of the leaves of C. rotundus had been explored for antidepressant and sedative-hypnotic activity. The results are as follows:
Acute toxicity test
After oral administration of C. rotundus extract at the doses of 500, 1000, and 2000 mg/kg, p.o., no toxicity and no significant changes in the BW between the control and treated group were demonstrated at these doses. This result indicates that the LD50 was higher than 2000 mg/kg.
Hole-board test is used to evaluate potential sedative-hypnotic activities of sample by analyzing exploratory behavior of mice. This test is popular for its simple procedure, and behavioral response of mice can easily be analyzed. Head-dipping characteristics of mice are associated to their emotional conditions. Decreasing number of head dipping behavior is a sign of sedative activity. Our present study results [Table 1] demonstrated that MECR did not show a dose-dependent reduction in head-dip response in the animals (4.01% and − 2.51% head-dip inhibition for 100 and 200 mg/kg doses, respectively) compared to control group recommending that the extract has no sedative activity. The perceived results from the treated groups were significantly different from that of the standard (diazepam treated) group [Table 1].
|Table 1: Effects of methanolic extract of Cyperus rotundus on hole-board test (n=5)|
Click here to view
Open field test
Results obtained from open field test showed that number of square crossed by MECR-treated groups (100 and 200 mg/kg BW) were almost similar to control group [Table 2]. Activity differences between diazepam-treated group and MECR-treated groups revealed that the plant extracts have no suppression effects on locomotors activity, which in turn is an indicative of its ineffectiveness against depression.
|Table 2: Effects of methanolic extract of Cyperus rotundus on open field test (n=5)|
Click here to view
Forced swimming test
From [Table 3], it was observed in FST that standard sample (nortriptyline) significantly decreased immobility time compared to the control group. Here, the doses of the MECR could not reduce immobility time significantly in comparison with control and standard group. However, the result obtained from both of the doses of plant extracts were almost similar to each other and control group.
|Table 3: Effects of methanolic extract of Cyperus rotundus on forced swimming test and tail suspension test (n=5)|
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Tail suspension test
In tail suspension test, both of the doses of plant extracts (100 and 200 mg/kg BW) could not significantly decrease immobility time in comparison with control group but nortriptyline showed significant reduction in immobility time in comparison with control group [Table 3]. Hence, both of the doses of methanolic extract of C. rotundus (MERC) showed insignificant inhibitory activity compared to the standard drug.
| Discussion|| |
In this investigation, we performed acute toxicity, sedative-hypnotic, and antidepressant activity test of C. rotundus plant extracts to evaluate its safety and possible neuropharmacological activity. Sample having sedative-hypnotic and antidepressant activity can be used to discover possible drug which can act against neurological disorders.
We evaluated acute toxicity in mice at the doses of 500, 1000, and 2000 mg/kg BW to measure its safety parameters in animal. After observing results for several days, we can say that this plant is safe to use and its LD50 is >2000 mg/kg.
Forced swimming and tail suspension tests are two well established test for observing antidepressant activity of sample. Decreasing immobility time is an indicative of antidepressant activity. In our study both of the tests showed that MECR in 100 and 200 mg/kg doses could not reduce immobility time significantly like standard sample. Immobility time observed in mice after treated with MECR were almost similar to the control group. However, the plant extracts failed to show any significant antidepressant activity in forced swimming and tail suspension test.
Hole cross and open field test were used to assess sedative-hypnotic behavior of plant extracts. A dose-dependent reduction in number of head dips in hole board test and number of square crossed in open field test is an evidence of sedative-hypnotic effects. Our study demonstrated that MECR could not notably reduce number head dips and number of square crossed in mice as compared to control and standard group. Results of both tests indicated an absence of sedative-hypnotic activity of our plant extracts.
| Conclusion|| |
This study results revealed that methanolic whole plant extracts of C. rotundus is safe to use in animal but had insignificant sedative-hypnotic and antidepressant potential in albino mice. Extensive study regarding the plant materials can further be undertaken to discover their other pharmacological efficacy and to rationalize their applications as traditional medicines.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3]