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| ORIGINAL ARTICLE |
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| Year : 2015 | Volume
: 9
| Issue : 1 | Page : 12-19 |
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An investigation to evaluate the analgesic and central nervous system depressant activities of Solanum nigrum (Linn.) in Homoeopathic potencies in experimental animal models
Echur Natarajan Sundaram1, Kushal Pal Singh2, Pratap Karnati Reddy3, Kainikkara Raven Janardanan Nair4, Anil Khurana2, Hari Singh2, Chaturbhuja Nayak2
1 Pharmacology Division, Central Research Institute (H), Noida, Uttar Pradesh, India
2 Central Council for Research in Homoeopathy, New Delhi, India
3 Department of Zoology, Osmania University, Hyderabad, Telangana, India
4 Central Research Institute (H), Kottayam, Kerala, India
| Date of Submission | 29-May-2014 |
| Date of Acceptance | 27-Nov-2014 |
| Date of Web Publication | 31-Mar-2015 |
Correspondence Address:
Echur Natarajan Sundaram
Central Research Institute (H), A?1/1, Sector ? 24, Noida ? 201 301, Uttar Pradesh
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0974-7168.154343
Background and Objective: In Homoeopathy, Solanum nigrum is clinically used in the treatment of ergotism, meningitis, irritation during dentition and some of the symptoms of neurological disorders but its Central Nervous System (CNS) potential has not been explored experimentally yet. Therefore, a preliminary study was conducted with an objective to evaluate the analgesic and CNS depressant effects of homoeopathic potencies of S. nigrum in experimental animal models.
Materials and Methods: The study was conducted in Wistar albino rats using a hot plate, ice plate and Randall-Selitto assay for analgesic; rota-rod and open field test for CNS depressant activities. The different potencies (3X, 6X, 12X and 30C) of Solanum nigrum were administered orally (0.5 ml/rat/day) for 30 days and response was assessed after 30 minutes of drug administration on 10 th , 20 th and 30 th day.
Results: The result shows that all the four potencies of Solanum nigrum has increased the latency time required to raise and lick the paws for thermal sensation on hot plate test and for cold sensation on ice plate test and also increased the degree of threshold pressure to mechanically induced pain on Randall-Selitto assay but depressed the motor coordination and locomotor activities.
Conclusion: The result obtained from this preliminary study suggests that homoeopathic preparation of Solanum nigrum in different potencies possess analgesic and CNS depressant activities. Further detailed investigations are required for its possible human use. Keywords: Albino rats, Analgesic, Central nervous system depressant activity, Motor coordination, Neurological disorders, Solanum nigrum
How to cite this article:
Sundaram EN, Singh KP, Reddy PK, Nair KJ, Khurana A, Singh H, Nayak C. An investigation to evaluate the analgesic and central nervous system depressant activities of Solanum nigrum (Linn.) in Homoeopathic potencies in experimental animal models. Indian J Res Homoeopathy 2015;9:12-9 |
How to cite this URL:
Sundaram EN, Singh KP, Reddy PK, Nair KJ, Khurana A, Singh H, Nayak C. An investigation to evaluate the analgesic and central nervous system depressant activities of Solanum nigrum (Linn.) in Homoeopathic potencies in experimental animal models. Indian J Res Homoeopathy [serial online] 2015 [cited 2023 Mar 22];9:12-9. Available from: https://www.ijrh.org/text.asp?2015/9/1/12/154343 |
| Introduction | |  |
Solanum nigrum commonly known as Kakmachi grows naturally throughout India. Solanum nigrum is an erect herb that grows about 30-45 cm in height [1] and demonstrates a diversity of therapeutic properties. It has been extensively used traditionally to treat various ailments such as pain, inflammation and fever. [2] In ethnomedicine, the plant is used as a remedy for cough, asthma, nasal catarrh, ophthalmic disorders, skin disorders, giddiness and nervous disorders. [3] The plant is also used in oriental systems of medicine as a diuretic and an antitumorigenic agent. [4]
Solanum nigrum possesses various compounds that are responsible for diverse activities. Its major active components are glycoalkaloids, glycoproteins, and polysaccharides. It also contains polyphenolic compounds such as gallic acid, catechin, protocatechuic acid, caffeic acid, epicatechin, rutin and naringenin. [5]
The fruit of Solanum nigrum has been found to possess antiulcer and antitumor effects in rats. [6] The extracts of plant exhibit antinociceptive, anti inflammatory [7] and antipyretic activity in rats [8] whereas its aqueous extract show anticonvulsant property in chicks, mice and rats. [9] Ethanol extract of Solanum nigrum has been reported to possess hepatoprotective activity against CCI 4 -induced hepatic damage in rats. [10] The ethanol extract of the fruit is also shown to possess potential Central Nervous System (CNS) depressant action. [11] Crude extract of Solanum nigrum leaves has been found to possess antioxidant effect in rats. [12]
In Homoeopathy, Solanum nigrum in different potencies used for treating ergotism, tetanic spasms and stiffness of whole body with mania, meningitis, irritation during dentition, chronic intestinal toxemia and other neurological disorders [13] . But the scientific evidence for its use in the treatment of neurological disorders as used in ethnomedicine was lacking. The present study was, therefore, carried out to assess the analgesic and CNS depressant activities of the homoeopathic potencies (3X, 6X, 12X and 30C) of Solanum nigrum in experimental animal models. The positive results in animal models, if obtained, can be made more meaningful by conducting human trials to prove its therapeutic effects on human brain.
| Materials and methods | | |
Collection of Plant Material
The whole plant of Solanum nigrum Linn. (Family: Solanaceae) was collected from Nilgiris hills, Tamil Nadu, India during the month of July and taxonomically identified/authenticated by the Botanist at Survey of Medicinal Plants Collection Unit, Udagamandalam, Tamilnadu.
Preparation and Selection of Medicines
Homoeopathic potencies of Solanum nigrum in 3X, 6X and 12X dilutions in Decimal Scale and 30C dilution in Centesimal Scale were prepared according to the standard procedures mentioned in Homoeopathic Pharmacopoeia of India and supplied by M/S. Bahola Laboratories, Puducherry, India from a single batch of whole plant. [14] The 3X, 6X, 12X and 3C dilutions of Solanum nigrum were selected in this study because these potencies are commonly prescribed by Homoeopathic practitioners for their clinical uses. [13]
Animals
Adult Wistar albino rats (120-140 g) procured from National Centre for Laboratory Animal Sciences, Hyderabad were used in the study. They were housed in polypropylene cages (47 cm × 34 cm × 20 cm) under standard laboratory conditions (12/12 hour, light/dark cycles and room temperature 25C ± 2C) and allowed free access to food and water ad-libitum. The animals were acclimatized to laboratory conditions for a period of 10 days before initiation of the study.
Experimental Design
The study protocol was approved by the Institutional Animal Ethics Committee (Reg. No. 383/01/a/CPCSEA), Department of Zoology, Osmania University, Hyderabad, where the study was conducted. A total of 180 rats were taken and divided into five batches (one batch for each study) of 36 each which were further divided into six groups of six each. The animals were accustomed to respective test procedures initially by subjecting them for test trials for three subsequent observations at 10-minute intervals on each day for 3 days before giving them any drug treatment. The test potencies (3X, 6X, 12X and 30C) of Solanum nigrum and vehicle (91.5% alcohol used as vehicle for preparation of different potencies of test drug) were diluted with distilled water in the ratio of 1:4 and kept as stock solutions. The animals were fed with 2 ml stock solution of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum that is, 0.5 ml (drug)/rat/day for 30 days. Vehicle and saline-treated rats were also maintained simultaneously along with drug-treated animals. The response of the drug was measured after 30 minutes of drug administration on 10 th , 20 th and 30 th day. Reading taken just before administration of the drug/vehicle/saline on first day of the study was considered as the initial control value in the same group for comparison.
Analgesic activity
Analgesic activity was studied by (i) hot plate (ii) ice plate and (iii) Randall-Selitto methods
Hot plate method
The hot plate latency assay was performed as per the method of Eddy et. al. [15] 30 minutes after the administration of drug, vehicle or saline, the rats were gently placed individually on a hot plate maintained at a constant temperature of 55C ± 2C and recorded the latency time (s). The time taken by the animals to lick the fore or hind paw or jump out of the hot plate was considered as latency (reaction) time. The reaction time noted just before administration of the drug, vehicle or saline on day 1 first day of the study was considered as the initial control values for comparison. A cut-off reaction time of 15 seconds was chosen in order to avoid physical injury to the animal. [16]
Ice plate method
Ice plate test was carried out as described by Sundaram et al.[17] After 30 minutes of drug, vehicle or saline administration, all rat, one at a time was gently placed on the ice cubes (0-4C) filled in a transparent container (20 cm × 20 cm × 20 cm) and covered with a plastic cover. The rat was visualized through the transparent wall and latency time (s) taken to lick the fore or hind paws to cold sensation was noted. The latency time of animal to cold sensation taken just before administration of drug, vehicle or saline on day first day of the study was considered as the initial control values for comparison. A cut-off reaction time was set at 15 seconds in order to avoid tissue damage to the animals due to frozen temperature.
Randall-Selitto method
The analgesic activity of the drug against mechanically induced pain was measured by Randall-Selitto assay. [18] After 30 minutes of drug, alcohol or saline administration, the rats were gently held in the hand. Afterward, the paw of the right foot of the rat was placed on the rubber base of the apparatus and pressure (in pounds; expressed in grams) was applied either on 2 nd -3 rd or 3 rd -4 th metatarsal region through a pointed tip and increased gradually until vocalization elicited which was considered as threshold pressure to mechanical induced pain. Threshold pressure to mechanical induced pain taken on first day just before the administration of drug, alcohol or saline was considered as the initial control values for comparison.
Central Nervous System depressant activity
Central Nervous System depressant activity was performed using (i) Rota-rod and (ii) open field methods
Rota-rod Test method
Motor coordination and grip strengths were measured using the automated rota-rod apparatus. [19] The rats capable of remaining on the rota-rod for 60 second or more, in three successive trials were selected for the study. After 30 minutes of drug, vehicle or saline administration, the rats were gently placed on the rotor with the body axis perpendicular to the rotor's long axis with the head directed opposite to the direction of rotating rod (5 rpm) and the fall-off time from the rod was noted for each rat. The control grip strengths of the rats were measured on first day just before administration of the drug, vehicle or saline for comparison.
Open field test method
For recording the locomotor activity of the rats, the Open Field Test was used. [20] The apparatus (96 cm × 96 cm × 6 cm) was made up of the wooden box and divided in to 36 equal squares which were painted alternatively with black and white colors. At the time of the experiment, it was illuminated with low-intensity diffuse light (40 W) placed at a height of 100 cm. The rats were placed gently in the center of the apparatus one after another and the number of squares crossed in 5 minutes was recorded before and 30 minutes after drug, vehicle or saline administration. The floor of the box was cleaned after every trial.
Statistical Analysis
The data were expressed as mean ± standard error of the mean. The difference between mean values of groups were statistically analyzed by student's' t- test. P <0.05 were considered significant. [21]
| Results | | |
Analgesic Activity
Hot plate test
[Table 1] represents the data for the analgesic effect of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum. The latency time required to raise and lick the hind paw for thermal stimulus was more or less same (3.38-3.67 seconds) in untreated (on first day before administration of drug/vehicle) and saline-treated animals when measured on different days of the experiment. On the other hand, the animals treated with different potencies of Solanum nigrum and vehicle at a dose of 0.5 ml/rat/day showed increase in the latency time (5.22-5.88 seconds) to thermal stimulus when measured after 30 minutes of drug/vehicle administration on 10 th day. The difference in the increase in latency time to hot sensation was significant (P < 0.05) with those groups which were treated with 3X and 6X potencies of Solanum nigrum. Thereafter, the increase in the duration of latency time tapered off gradually on 20 th day and 30 th day of treatment [Figure 1]. | Table 1: Analgesic effect of S. nigrum (0.5 ml/rat/day) on a hot plate test
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Ice plate test
[Table 2] represents the data for the analgesic effect of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum. The latency time required to raise and lick the hind paw for cold sensation was more or less same (5.80-6.09 seconds) in untreated and saline-treated animals when measured on different days of the experiment whereas Solanum nigrum in different potencies and vehicle at a dose of 0.5 ml/rat/day had significantly increased the latency time (7.31-8.09 seconds) on 10 th day which was gradually tapered off on 20 th day and 30 th day of treatment. The difference was significant (P < 0.05) only with those rats treated with 3X and 6X when compared to initial latency time taken just before administration of drug on first day of the study [Figure 2]. | Table 2: Analgesic effect of S. nigrum (0.5 ml/rat/day) on ice plate test
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Randall-Selitto assay
The results of the analgesic effect of different potencies of Solanum nigrum are presented in [Table 3]. The quantum of threshold pressure required to elicit vocalization to applied mechanical pain was more or less same (131.33-133.66 g) on first day before administration of drug, vehicle or saline and 30 minutes after the administration of saline on different days of experimentation. There was an increase in the quantum of applied threshold pressure (145.00-148.66 g) required to elicit vocalization to mechanical pain when measured 30 minutes after the administration of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum and vehicle at a dose of 0.5 ml/rat/day on 10 th day. The difference was significant (P < 0.05) only with those rats treated with 3X and 30C potencies when compared to that of normal saline-treated rats. Afterward, the increase in the quantum of threshold pressure required to elicit vocalization to applied mechanical pain tapered off gradually on 20 th day and 30 th day of experiments [Figure 3]. | Table 3: Analgesic effect of S. nigrum (0.5 ml/rat/day) on Randall-Selitto test
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Central Nervous System depressant activity
Rota-rod test
The results of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum on motor coordination activity of rats are summarized in [Table 4]. Untreated rats and rats administered with saline stayed on the rotating rod on an average of 49.07-51.55 seconds whereas, rats treated with different potencies of Solanum nigrum and vehicle at a dose of 0.5 ml/rat/day fell between 32.81 and 38.05 seconds when they were subjected to test on 10 th day of experiment after 30 minutes of drug administration. The decrease in grip strength was significant (P < 0.05) only in those rats treated with 3X and 6X potencies of Solanum nigrum. Afterward, there was a progressive increase in the grip strength of drug/vehicle treated rats as they stayed comparatively for more time on the rotating rod when tested on 20 th and 30 th day of experiment though stay on rotating rod was still less than that of control initial values observed in the same group on first day just before administration of drug or with that of saline administered rats [Figure 4]. | Table 4: Behavioral effect of S. nigrum (0.5 ml/rat/day) on rota-rod test
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Open field test
The results of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum and vehicle on locomotor activity of rats are presented in [Table 5]. Control untreated rats and rats administered normal saline crossed on an average of (63.00-65.66) squares in 5 minutes. On the other hand, there was a decrease (46.00-47.33 squares in 5 minutes) in locomotor activity of rats administered Solanum nigrum and vehicle at a dose of 0.5 ml/rat/day when tested after 30 minutes of drug administration on 10 th day of experiment. The decrease in locomotor activity was significant (P < 0.05) only with those rats treated with 6X and 12X potencies of Solanum nigrum when compared to initial latency time taken just before administration of drug on first day of the study [Figure 5]. | Table 5: Behavioral effect of Solanum nigrum (0.5 ml/rat/day) in open field test
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| Discussion and conclusion | | |
Solanum nigrum is used as homoeopathic drug to treat various ailments including many symptoms of neurological disorders clinically, but has not garnered the attention to verify its use experimentally. [15] Therefore, the present preliminary study was carried out for the first time with an objective to assess the analgesic activity using hot plate, ice plate and Randall-Selitto assays and CNS depressant activities using Rota-rod Test (for motor coordination) and Open Field Test (for locomotor activity) of homoeopathic potencies of Solanum nigrum (3X, 6X, 12X and 30C) in albino rats.
The data for the analgesic effect showed that all the four (3X, 6X, 12X and 30C) potencies of Solanum nigrum has increased the latency time required to raise and lick the hind paw for thermal sensation on hot plate assay and for cold sensation on ice plate assay and also increased the quantum of threshold pressure to mechanical induced pain on Randall-Selitto assay when measured 30 minutes after the administration of different potencies (3X, 6X, 12X and 30C) of Solanum nigrum on 10 th day. These effects were gradually tapered off on 20 th day and 30 th day of treatment.
The data obtained for CNS depressant effect showed that different potencies (3X, 6X,12X and 30C) of Solanum nigrum has significantly decreased motor co-ordination on Rota-rod Test and locomotor activity in Open Field Test. The depressant effect was the maximum on 10 th days and slowly decreased thereafter very similar to that of analgesic response. The level of significance varied not only between the potencies of the drug but also between different sets of experiments as compared to the values of saline group or initial values taken just before administration of drug on first day of the study.
Increased in the latency time to noxious thermal stimulus and/or cold sensation, and increased in the quantum of threshold pressure to mechanically induced pain on drug treatment are the clear indication that the drug possess analgesic effect. [22] Similarly, decreased motor coordination and locomotor activity of the drug is the sign of CNS depression. [23] Wearing off the analgesic and depression on prolonged and continuous use of the drug may be either due to decreased sensitivity of the CNS or due to increased metabolizing enzymatic activity in the liver.
The present preliminary study demonstrates that the Homoeopathic potencies (3X, 6X, 12X and 30C) of Solanum nigrum possess analgesic and CNS depressant effect. However, further detailed investigations are needed for its possible clinical use.
| Acknowledgment | | |
The authors are thankful to Head, Department of Zoology, Osmania University, Hyderabad for extending laboratory facilities for conducting the study. The authors acknowledge the financial support from Ministry of AYUSH under collaborative study and Director General, CCRH for providing the support.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]
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