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ORIGINAL ARTICLE
Year : 2012  |  Volume : 1  |  Issue : 2  |  Page : 94-98

Evaluation of centrally acting skeletal muscle relaxant activity of aqueous extract of Cinnamomum zeylanicum bark in albino mice


Department of Pharmacology, Mamata Medical College, Khammam, Andhra Pradesh, India

Date of Web Publication11-Jul-2012

Correspondence Address:
Jayasree Tirumalasetty
Department of Pharmacology, Mamata Medical College, Khammam - 507002, Andhra Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2277-8632.98345

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  Abstract 

Background: Cinnamomum Zeylanicum is traditionally used in various diseases because of its medicinal properties. Cinnamon is a spice obtained from the inner bark of several trees from the genus Cinnamomum that is widely used as a spice. Cinnamomum Zeylanicum is principally employed in cookery as a condiment and flavoring material. One of its uses is in musculoskeletal disorder.
Aims: The study aims to evaluate the skeletal muscle relaxant activity of the Aqueous Extract of Cinnamomum Zeylanicum Bark (AECZB) in albino mice in Comparison with Diazepam.
Materials and Methods: Thirty Swiss albino mice (SWR) aged six to seven weeks, of either sex, weighing about 25 - 30 g, were taken, and after acute toxicity studies three different doses were selected. The animals were divided into five different groups. The first group was kept as the control (Normal Saline), second as the standard (Diazepam) and the remaining three groups as Test I, Test II, and Test III, and given different doses of the AECZB. Skeletal muscle relaxant activity (motor coordination) on Rotarod and Locomotor activity on Photoactometer were performed. The results were expressed as mean ± SD. Statistical analysis was carried out by using ANOVA, followed by Dunnet's multiple comparison tests, using primer of biostatistics McGraw-Hill software version 5.0.0.0 (2011). P-values < 0.05 were considered significant.
Results: The result from the Actophotometer test and Rotarod test showed that the extract significantly reduced the motor coordination of the tested animals.
Conclusions: Our data indicates that AECZB possesses sedative and skeletal muscle relaxant activities.

Keywords: Actophotometer, cinnamomum zeylanicum, rota-rod, soxhlet′s apparatus, muscle relaxant


How to cite this article:
Tirumalasetty J, Rasamal K, Nutalapati C, Samreen S. Evaluation of centrally acting skeletal muscle relaxant activity of aqueous extract of Cinnamomum zeylanicum bark in albino mice. J NTR Univ Health Sci 2012;1:94-8

How to cite this URL:
Tirumalasetty J, Rasamal K, Nutalapati C, Samreen S. Evaluation of centrally acting skeletal muscle relaxant activity of aqueous extract of Cinnamomum zeylanicum bark in albino mice. J NTR Univ Health Sci [serial online] 2012 [cited 2019 Oct 24];1:94-8. Available from: http://www.jdrntruhs.org/text.asp?2012/1/2/94/98345


  Introduction Top


Cinnamomum Zeylanicum (CZ) is commonly known as cinnamon. Cinnamon oil has been reported to possess antimicrobial and anti-inflammatory [1] actions. Pharmacological experiments suggest that the cinnamon-derived dietary factor, cinnamic aldehyde, activates the Nrf2-dependent antioxidant response in human epithelial colon cells, representing an experimental chemopreventive dietary factor targeting colorectal carcinogenesis. [2] The anti-melanoma activity of cinnamic aldehyde can be observed in cell culture and a mouse model of human melanoma. [3] A 2011 study isolated a substance (CEppt) in the cinnamon plant, which inhibited the development of Alzheimer's in mice. [4] CEppt, an extract of cinnamon bark, seems to treat a mouse model of Alzheimer's disease. [5] Another study found that CZ exhibited anti-inflammatory activity, by inhibiting nitric oxide production via inhibition of NF-kappa B. [6] Its anti-inflammatory properties reduced the stiffness and pain associated with this musculoskeletal disorder. This study is taken to elucidate the skeletal muscle relaxant action of CZ.


  Materials and Methods Top


Collection of the Bark

The study was conducted during the period January 2012 to February 2012. The bark of Cinnamomum Zeylanicum was obtained from a local supermarket. The identification and authentication of the bark was done at the Department of Botany, Government Degree College. The aqueous extract of powdered bark was prepared using the soxhlet's apparatus in the Department of Pharmacology. The extract was dried under vacuum, stored at room temperature, and protected from direct sunlight.

Animals

Swiss albino mice (SWR) aged six to seven weeks of either sex weighing about 25 - 30 g, were obtained from the Central Animal House. The animals were fed a Purina Chow diet, with water ad libitum, and were maintained under standard conditions of temperature, humidity, and light (12 hours light / 12 hours dark cycle). The experiment complied with the guidelines for animal experimentation of our laboratory and was approved by the Institutional Animal Ethics Committee (IAEC); Registration number 285 / CPCSEA. The guidelines for the investigation of experiments in conscious animals were followed in all tests.

Drugs and Chemicals

Diazepam (Lupin Laboratories Ltd., India), 10 mg / kg and normal saline (0.9% NaCl solution) were administered in a volume of 10 ml / kg. The extracts were suspended in distilled water and subjected to muscle relaxant activity using the Rotarod apparatus and Actophotometer. The extracts were administered orally (p. o.) in a volume of 10 ml / kg of body weight, in doses of 50 mg, 100 mg, and 200 mg / kg

Acute Toxicity

A total of 35 mice were randomly allotted to one control and six treatment groups. The animals were fasted overnight, prior to the experimental procedure. The method of up-and-down or staircase was used to determine the dose. [7],[8] The procedure was followed as per the Organization for Economic Co-operation and Development (OECD) 423 guidelines. The extract in each case was administered orally in three doses: 0.5 g / kg, 1.0 g / kg, and 3 g / kg. The animals were observed for 24 hours for signs of toxicity and mortality. [9] In the acute toxicity tests, C. zeylanicum treated animals exhibited no alarming signs of toxicity. Only at the 3 g / kg dose level there was some decreased locomotor activity observed. This finding verified the earlier report where C. zeylanicum was described as possessing narcotic effects at high doses. [10],[11]

Selection of Dose for Pharmacological Screening

The aqueous extract of Cinnamomum Zeylanicum was found to be non-toxic up to a dose of 2000 mg / kg and did not cause death, therefore it was considered to be safe. Hence, one-tenth of this dose, that is, 200 mg / kg body weight and half of the one-tenth dose, that is, 100 mg / kg, as also half of this, that is, 50 mg / kg, were used for the elucidation of muscle relaxant activity.

Experimental Design

Group I - Control Rats (Normal saline 10 ml / kg)

Group II - Standard (Diazepam 10 mg / kg)

Group III -AECZB 50 mg / kg

Group IV - AECZB 100 mg / kg

Group V - AECZB 200 mg / kg

(I) Skeletal muscle relaxant activity (motor coordination) [7],[12] The mice were divided into five groups consisting of six animals each. Group I served as the control, which received Normal saline 10 ml / kg, Group II received the standard drug Diazepam, at a dose of 10 mg / kg, p.o., Group III, IV, and V received the aqueous extract of Cinnamomum Zeylanicum bark orally at a dose of 50, 100, and 200 mg / kg. The animals remained on Rotarod (25 rpm) for five minutes or more after low successive trials were included in the study. After the administration of control, standard, and test material, the fall off time from the rotating rod was noted after 30 minutes. The difference in the fall off time from the rotating rod between the control and the treated mice was taken as an index of muscle relaxation.

(II) Locomotor activity: The spontaneous locomotor activity was assessed with the help of a photoactometer. [13] Each animal was observed for a period of five minutes in a square closed field arena (30 × 30 × 30 cm) equipped with six photocells in the outer wall. Interruptions of photocell beams (locomotor activity) were recorded by means of a six digits counter.

To see the locomotor activity, the Actophotometer was turned on and each mouse was placed individually in the activity cage for five minutes. The basal activity score for all the animals was noted. Control normal saline, Standard Diazepam, and three different doses of aqueous extract of Cinnamomum Zeylanicum bark were given orally and after one hour of re-testing, and the activity score for five minutes was observed. The difference in the activity, before and after drug administration, was noted. The percentage decrease in motor activity was calculated.

Statistical Analysis

The results were expressed as mean ± SD. Statistical analysis was carried out by using the Analysis of Variance (ANOVA) followed by Dunnet's multiple comparison tests using primer of windows McGraw-Hill software version 5.0.0.0 (2011). P-values < 0.05 were considered significant.


  Results Top


Rotarod Test

For muscle relaxation

In this test, AECZB (50 mg / kg, 100 mg / kg, and 200 mg / kg) showed highly significant reduction in the time spent by the animals on the revolving rod when compared to the control (P < 0.01). The standard drug (Diazepam) also showed a highly significant effect when compared to the control (P < 0.01). Low dose of AECZB (50 mg / kg) showed a significant effect (P value < 0.05) [Table 1]. Three different doses of AECZB (50, 100, and 200 mg / kg p.o.) showed a dose-dependent increase in muscle relaxation, that is, 62.3, 77.7, and 79%, respectively, when compared to the control. Maximum muscle relaxation was observed with 200 mg / kg of Cinnamomum zeylanicum. The result from the Rotarod test showed that the extract significantly reduced the motor coordination of the tested animals.
Table 1: Effect of AECZB on the locomotor activity on the actophotometer and muscle Coordination on the rotarod apparatus

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Actophotometer

Test for locomotor activity: The percentage of reduction in the locomotor activity with Diazepam (10 mg / kg, p.o.) after 30 minutes was 89.5%, that is, there was a highly significant (P < 0.000) decrease in locomotor activity compared to the control, whereas, three different doses of AECZB (50,100, and 200 mg / kg, p.o.) showed a dose-dependent decrease in the locomotor activity, that is, 62.3, 77.7, and 79%, respectively, when compared to the control. Maximum muscle relaxation was observed with 200 mg / kg of Cinnamomum Zeylanicum. The values were highly significant (P < 0.000) [Table 1].


  Discussion Top


Cinnamon is one of the world's oldest spices. Cinnamon comes from the inner bark of the shoots of a tree (Cinnamomum Zeylanicum) that grows predominantly in India, China, and Sri Lanka. The fruit and coarser pieces of bark, when boiled, yield fragrant oil. Cinnamon is aromatic and one of the best tasting spices. In recent years scientists have discovered that cinnamon extract has a strong antioxidant activity and has the potential to help maintain healthy blood sugar and cholesterol levels. One of the main uses of C. zeylanicum in herbal medicine is to improve insulin resistance and blood sugar levels, and to help balance the metabolism of individuals with a metabolic syndrome. C. zeylanicum has been reported to help improve insulin sensitivity, in both laboratory and human studies. [14],[15],[16],[17],[18]

The herb stimulates the circulation, especially to the fingers and toes and has been used for arthritis. Cinnamon is also a traditional remedy for aching muscles and other symptoms of viral conditions such as colds and flu. The cinnamon bark contains volatile oils, cinnamaldehyde, eugenol, transcinnamic acids; phenolic compounds, condensed tannins, catechins, and proanthocyanidins; monoterpenes, and sesqiterpenes, (pinene); calcium monoterpenes oxalate; gum; mucilage; resin, starch, sugars, and traces of coumarin. [19] The bark contains dimeric, trimeric, and higher oligomeric proanthocyanidins with doubly linked bis-flavan-3-ol units in the molecule. [20] The bark of Cinnamomum zeylanicum was found to contain a major phenolic metabolite of doubly linked proanthocyanidins. C. zeylanicum bark extract was found to have anti-inflammatory activity. Cinnamomum species including Cinnamomum osmophloeum [21] Cinnamomum camphora [22] Cinnamomum insularimontanum [23] and Cinnamomum cassia[24] have also been demonstrated to have anti-inflammatory properties. Cinnamon possesses chemopreventive, antispasmodic, sedative, hypothermic, choleretic, antibacterial, antifungal, antipyretic, antiviral, antiplatelet, antiseptic, lipolytic, anesthetic, cytotoxic, anodyne, and hypolipidemic properties, and also stimulates the immune system, which may be useful adjuncts in helping to reduce the risk of cardiovascular disease and cancer.

In evaluation of the efficacy and safety of Rumalaya gel, in the management of acute and chronic inflammatory musculoskeletal disorders, [25] the beneficial effects of Cinnamomum zeylanicum, such as, reduction of swelling associated with inflammatory conditions, shortening of recovery time, and increase of mobility in the joints were observed. In the 1980s, Japanese researchers found that the active constituent, cinnamaldehyde, acted as a sedative and analgesic. [26]


  Conclusion Top


The result of our study shows a dose-dependent decrease in locomotor activity when compared to the control, on the Actophotometer. Maximum muscle relaxation was observed with 200 mg / kg of Cinnamomum Zeylanicum. The Rotarod test showed highly significant reduction in the time spent by the animals on the revolving rod when compared to the controls. The result from the Actophotometer test and Rotarod test showed that the extract significantly reduced the motor coordination of the tested animals. In conclusion, our data indicates that AECZB possesses sedative and skeletal muscle relaxant activities. The AECZB contains volatile oils, cinnamaldehyde, eugenol, and transcinnamic acids, and phenolic compounds, which are probably responsible for the actions. [26] Further studies have to be conducted to elucidate the above-mentioned properties.

 
  References Top

1.Gomez Flores R, Hernandez Martinez H, Tamez-Guerra P, Tamez-Guerra R, Quintanilla-Licea R, Monreal Cuevas E, et al. Antitumor and immunomodulating potential of Coriandrum sativum, Piper nigrum and Cinnamomum zeylanicum. J Nat Prod 2010;3:54-63.  Back to cited text no. 1
    
2.Wondrak GT, Villeneuve NF, Lamore SD, Bause AS, Jiang T, Zhang DD. The cinnamon-derived dietary factor cinnamic aldehyde activates the Nrf2-dependent antioxidant response in human epithelial colon cells. Molecules 2010;15:3338-55.  Back to cited text no. 2
    
3.Cabello CM, Bair WB, Lamore SD, Ley S, Bause AS, Azimian S, et al. The cinnamon-derived Michael acceptor cinnamic aldehyde impairs melanoma cell proliferation, invasiveness, and tumor growth. Free Radic Biol Med 2009;46:220-31.  Back to cited text no. 3
    
4.Haaretz.com [Internet]. Jewish World News. Available from: http://www.haaretz.com/print-edition/news/tau-finds-cinnamon-could-combat-alzheimer-s-1.366678 [Last Accessed on 2011 May 24].  Back to cited text no. 4
    
5.Frydman-Marom A, Levin A, Farfara D, Benromano T, Scherzer-Attali R, Peled S, et al. Orally administrated cinnamon extract reduces â-amyloid oligomerization and corrects cognitive impairment in Alzheimer's disease animal models. PLoS One 2011;6:e16564.  Back to cited text no. 5
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7.Ghosh MN. Fundamentals of experimental pharmacology. 2 nd ed. Kolkata: Scientific Book Agency; 1984. p. 156.  Back to cited text no. 7
    
8.Perez GR, Perez LJ, Garcia DL, Sossa MH. Neuropharmacological activity of Solanum nigrum fruit. J Ethnopharmacol 1998;62:43-8.  Back to cited text no. 8
    
9.Lipnick RL, Cotruvo JA, Hill RN, Bruce RD, Stitzel KA, Walker AP, et al. Comparison of the up-and-down, conventional LD50, and fixed-dose acute toxicity procedures. Food Chem Toxicol 1995;33:223-31.   Back to cited text no. 9
    
10.Perry LM, Metzger J, Medicinal plants of southeast Asia. Cambridge: The MIT Press; 1980. p. 198-9.  Back to cited text no. 10
    
11.Akira T, Tanaka S, Tabata M. Pharmacological studies on the antiulcergenic activity of Chinese Cinnamon. Planta Med 1986;6:440-3.  Back to cited text no. 11
    
12.Al-Nagger TB, Gómez-Serranillos MP, Carretero MP, Villar AM. Neuropharmacological activity of N.sativa L. J Ethnopharmacol 2003;88:63-8.  Back to cited text no. 12
    
13.Vogel HG, Vogel WH. Drug discovery and evaluation. 2 nd ed. Berlin, Heidelberg, New York: Springer-Verlag; 2002.  Back to cited text no. 13
    
14.Jarvill-Taylor KJ, Anderson RA, Graves DJ. A hydroxychalcone derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes. J Am Coll Nutr 2001;20:327-36.  Back to cited text no. 14
    
15.Solomon TP, Banning AK. Effects of short-term cinnamon ingestion on in vivo glucose tolerance. Diabetes Obes Metab 2007;9:895-901.  Back to cited text no. 15
    
16.Mang B, Wolters M, Schmitt B, Kelb K, Lichtinghagen R, Stichtenoth DO, et al. Effects of a cinnamon extract on plasma glucose, HbA, and serum lipids in diabetes mellitus type 2. Eur J Clin Invest 2006;36:340-4.  Back to cited text no. 16
    
17.Pham AQ, Kourlas H, Pham DQ. Cinnamon supplementation in patients with type 2 diabetes mellitus. Pharmacotherapy 2007;27:595-9.  Back to cited text no. 17
    
18.Khan A, Safdar M, Ali Khan MM, Khattak KN, Anderson RA. Cinnamon improves glucose and lipids of people with type 2 diabetes. Diabetes Care 2003;26:3215-8.  Back to cited text no. 18
    
19.Duke JA. Handbook of medicinal herbs. Boca Raton, Fia: CRC Press Inc.; 1985. p. 33-4.  Back to cited text no. 19
    
20.Fang SH, Rao YK, Tzeng YM. Inhibitory effects of flavonol glycosides from Cinnamomum osmophloeum on inflammatory mediators in LPS / IFN-gamma-activated murine macrophages. Bioorg Med Chem 2005;13:2381-8.  Back to cited text no. 20
    
21.Tung YT, Chua MT, Wang SY, Chang ST. Anti-inflammation activities of essential oil and its constituents from indigeno cinnamon (Cinnamomum osmophloeum) twigs. Bioresour Technol 2008;99:3908-13.  Back to cited text no. 21
    
22.Lee HJ, Hyun EA, Yoon WJ, Kim BH, Rhee MH, Kang HK, et al. In vitro anti-inflammatory and anti-oxidative effects of Cinnamomum camphora extracts. J Ethnopharmacol 2006;103:208-16.  Back to cited text no. 22
    
23.Li TJ, Qiu Y, Mao JQ, Yang PY, Rui YC, Chen WS. Protective effects of Guizhi-Fuling-Capsules on rat brain schemia / reperfusion injury. J Pharmacol Sci 2007;105:34-40.  Back to cited text no. 23
    
24.Lin CT, Chen CJ, Lin TY, Tung JC, Wang SY. Anti-inflammation activity of fruit essential oil from Cinnamomum insularimontanum Hayata. Bioresour Technol 2008;99:8783-7.  Back to cited text no. 24
    
25.Singh RB, Giri S, Kolhapuri SA. Evaluation of efficacy and safety of Rumalaya gel in the management of acute and chronic inflammatory musculoskeletal disorders: An open, prospective, non-comparitive, phase III clinical trial. Indian J Clin Pract 2005;15:1-15.  Back to cited text no. 25
    
26.Chevallier A. The encyclopedia of medicinal plants. US: DK Pub.; 1996.  Back to cited text no. 26
    



 
 
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