IPSIndian Journal of Pharmacology
Home  IPS  Feedback Subscribe Top cited articles Login 
Users Online : 11341 
Small font sizeDefault font sizeIncrease font size
Navigate Here
  Next article
  Previous article 
  Table of Contents
Resource Links
   Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
   Article in PDF (243 KB)
   Citation Manager
   Access Statistics
   Reader Comments
   Email Alert *
   Add to My List *
* Registration required (free)

In This Article
   Article Tables

 Article Access Statistics
    PDF Downloaded422    
    Comments [Add]    
    Cited by others 11    

Recommend this journal


Year : 2006  |  Volume : 38  |  Issue : 2  |  Page : 131-132

Antioxidant potential of methanolic extract of Dolichos biflorus Linn in high fat diet fed rabbits

1 Department of Pharmacy, Annamalai University, Annamalai Nagar, India
2 Division of Biochemistry, Rajah Muthiah Medical College, Annamalai University, Annamalai Nagar-608002, India

Correspondence Address:
A Kottai Muthu
Department of Pharmacy, Annamalai University, Annamalai Nagar
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0253-7613.24620

Rights and Permissions

How to cite this article:
Muthu A K, Sethupathy S, Manavalan R, Karar P K. Antioxidant potential of methanolic extract of Dolichos biflorus Linn in high fat diet fed rabbits. Indian J Pharmacol 2006;38:131-2

How to cite this URL:
Muthu A K, Sethupathy S, Manavalan R, Karar P K. Antioxidant potential of methanolic extract of Dolichos biflorus Linn in high fat diet fed rabbits. Indian J Pharmacol [serial online] 2006 [cited 2023 Sep 29];38:131-2. Available from: https://www.ijp-online.com/text.asp?2006/38/2/131/24620

Dolichos biflorus Linn (Fabaceae), is commonly known as Kollu in Tamil and horse gram in English. It has been reported to lower lipids in rats.[1] A high fat diet induces oxidative stress in the cells by producing reactive oxygen species. Therefore, in this study, the influence of the Dolichos biflorus extract on high fat diet (HFD) induced oxidative stress in rabbits, has been investigated.

Whole plants of D. biflorus were collected from Sankaran Koil, Tirunelveli district of Tamilnadu, India. Taxonomic identification was made by the Botanical Survey of Medicinal Plant Unit, Siddha, Government of India, Palayamkottai, Tamilnadu. Four month-old whole plants were dried in the shade, segregated, and pulverized by a mechanical grinder and passed through a 40 mesh sieve. The powder was extracted by methanol in Soxhlet apparatus by continuous hot percolation method. After filtration through Whatmann filter paper No 40, the filtrate was vacuum dried at 35 to 40oC. The extracts were stored in screw cap vials at 4oC until further use. The extractive value of the methanolic extract was 8.13% w/w. The methanolic extract of D.biflorus was subjected to preliminary phytochemical screening to find out the presence of active principles. The extracts were suspended in 2% tween 80.

New Zealand white rabbits, weighing 900-1050 g were procured from the central animal house, Rajah Muthiah Medical College, Annamalai University. The animals were kept in cages, 2 per cage, with 12:12 h light/dark cycle at 25 + 2oC. The animals were maintained on their respective diets and water ad libitum . Animal ethical committee's clearance was obtained for the study.

Rabbits were divided into the following five groups, with six rabbits in each group:

Group I : (Control): Standard chow diet.

Group II : High fat diet (HFD)

Group III : High fat diet plus methanolic extract of D.biflorus (dose I -200 mg/kg body weight)

Group IV : High fat diet plus methanolic extract of D.biflorus (dose II -400 mg/kg body weight)

Group V : High fat diet plus standard drug atorvastatin (1.2 mg/kg body weight).

The compositions of the two diets were as follows:

Control diet

Wheat flour 22.5%, roasted Bengal gram powder 60%, skimmed milk powder 5%, casein 4%, refined oil 4%, salt mixture with starch 4%, and vitamin and choline mixture 0.5%.

High fat diet

Wheat flour 20.5%, roasted Bengal gram 52.6%, skimmed milk powder 5%, casein 4%, refined oil 4%, coconut oil 9%, salt mixture with starch 4%, vitamin and choline mixture 0.5%, and cholesterol 0.4%.

Rabbits in groups III and IV were orally fed the methanolic extracts of D. biflorus dose I and dose II, respectively, and rabbits in group V were fed standard drug atorvastatin. The D. biflorus extracts and atorvastatin were suspended in 2% tween 80 separately and fed to the respective groups of rabbits by oral intubation. At the end of 11 weeks, all the animals were sacrificed by cervical decapitation after overnight fasting.

Portions of the tissues from liver, heart, and aorta were blotted, weighed, and homogenized with methanol (3 volumes). The lipid extract obtained by the method of Folch et al .[2] was used for the estimation of thiobarbituric acid reactive substances[3] (TBARS). Another portion of the tissues was homogenized with phosphate buffer saline and used for the estimation of reduced glutathione[4] (GSH), catalase[5] (CAT), and superoxide dismutase[6](SOD).

Results were expressed as mean + SE of 6 rabbits in each group. One-way analysis of variance (ANOVA) with Scheffe's multiple comparisons test, were used to determine the statistical significance. P<0.05 was considered significant.

The average body weight was found to have increased in high fat diet fed rabbits compared with those in control group. After administration of two doses of D.biflorus extract it was found to have decreased. [Table - 1] TBARS had significantly increased and GSH had significantly decreased in liver, heart, and aorta of rabbits fed HFD ([Table - 1]; Group II) compared with those in control group I. Administration of D.biflorus extract significantly lowered the level of TBARS and enhanced the level of GSH. Higher dose of the plant extract was found to be more effective and showed comparable results with standard drug atorvastatin on these two parameters. Activities of antioxidant enzymes, that is, SOD and CAT in different groups are given in [Table - 2]. These two enzymes showed a marked reduction in activity in liver, heart, and aorta of rabbits in group II which had been fed HFD. Supplementation of D.biflorus extract with HFD significantly improved the activities of SOD and CAT in the above tissues of rabbits in groups III and IV as compared with group II. Preliminary phytochemical study shows that methanolic extract of the experimental plant contains phytoconstituents, such as, alkaloids, steroids, flavonoids, and isoflavone.

Elevated levels of TBARS in liver, heart, and aorta in group II rabbits are a clear manifestation of excessive formation of free radical and activation of lipid peroxidation. The significant decline in the level of TBARS, in rabbits administered with D.biflorus (Groups III and IV), unveils the antioxidant potential of the D.biflorus extract. Both doses of the D.biflorus extract (groups III and IV) help to restore the GSH levels near normal. The higher dose has more effect, which was comparable to atorvastatin. Increase in GSH concentration in animals treated with D.biflorus extract may be due to increased activity of the enzyme, glutathione reductase, which catalyses the conversion of oxidized glutathione to reduced glutathione in liver. It may also be due to enhanced synthesis/transport of GSH.

Restoration of the activities of SOD and CAT to near normal, observed in tissues of rabbits supplemented with D.biflorous may be due to the removal of toxic intermediates by the plant extract in HFD fed rabbits. The components of the plant extract may also directly activate the antioxidant enzymes.[7] It is concluded that administration of D.biflorus manifests a protective action against HFD induced oxidative stress in different tissues in rabbits. However, further studies are needed to isolate the active principles, elucidate their structures, and determine their pharmacological activities.

  References Top

1.Kottai Muthu A, Sethupathy S, Manavalan R, Karar PK. Hypolipidemic effect of methanolic extract of Dolichos biflorus Linn in high fat diet fed rats . Indian J Exp Biol 2005;43:522-5.  Back to cited text no. 1    
2.Folch J, Lees M, Sloane GH. A simple method for the isolation and purification of total lipids from animals tissues. J Biol Chem 1957;226:497-9.  Back to cited text no. 2    
3.Nichans WH, Samulelson B. Formation of malondialdehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur J Biochem 1968;6:126-30.  Back to cited text no. 3    
4.Ellmann GL. Tissue sulfahydryl groups. Arch Biochem Biophys 1959;82:70-7.  Back to cited text no. 4    
5.Sinha AK. Colorimetric assay of catalase. Annal Biochem 1972;47:389-94.  Back to cited text no. 5  [PUBMED]  
6.Kakkar P, Das B, Visvanathan PN. A modified spectrophotometric assay of SOD. Indian J Biochem Biophys 1984;21:130-2.  Back to cited text no. 6    
7.Jeyakumar SM, Nalini N, Venugopal P Menon. Antioxidant activity of ginger (zingiber officinale Rose) in rats fed a high fat diet . Med Sci Res 1999;27:341-4.  Back to cited text no. 7    


[Table - 1], [Table - 2]

This article has been cited by
1 Effect of starter culture on underutilized millet-legume based Indian traditional fermented product in streptozotocin-induced diabetic rats
Palanisamy Bruntha Devi, Suresh Rajendran
Journal of Agriculture and Food Research. 2023; 11: 100483
[Pubmed] | [DOI]
2 Influence of processing methodology on phenolic acid content, fatty acid profile and oxidative stability of Macrotyloma uniflorum
Rahul Vashishth, A. D. Semwal, A. Padmashree, Mahadeva Naika, G. K. Sharma
Journal of Food Science and Technology. 2020; 57(8): 2894
[Pubmed] | [DOI]
3 Pharmacognostic and phytochemical evaluation of Dolichos biflorus Linn.
Shashi Alok,Sanjay Kumar Jain,Amita Verma,Mayank Kumar
Asian Pacific Journal of Tropical Disease. 2014; 4: S97
[Pubmed] | [DOI]
4 Chemical composition of ethanol extract of Macrotyloma uniflorum (Lam.) Verdc. using GC-MS spectroscopy
Sneha Das,Neeru Vasudeva,Sunil Sharma
Organic and Medicinal Chemistry Letters. 2014; 4(1): 13
[Pubmed] | [DOI]
5 Role of Antioxidant Herbal Drugs in Renal Disorders: An Overview
Rasikh Javaid,Mohd Aslam,Qudsia Nizami,Raheela Javaid
Free Radicals and Antioxidants. 2012; 2(1): 2
[Pubmed] | [DOI]
6 Chemomodulatory effect of Dolichos biflorus Linn. on skin and forestomach papillomagenesis in Swiss albino mice
Nanta, R., Kale, R.K.
Indian Journal of Experimental Biology. 2011; 49(7): 483-490
7 Antioxidant effect of triticum aestivium (wheat grass) in high-fat diet-induced oxidative stress in rabbits
Sethi, J., Yadav, M., Dahiya, K., Sood, S., Singh, V., Bhattacharya, S.B.
Methods and Findings in Experimental and Clinical Pharmacology. 2010; 32(4): 233-235
8 Dolichos biflorus Linn attenuate progression of renal damage in alloxan-induced diabetic rats
G.U. Chidrewar, V.S. Mane, M.M. Ghaisas, A.D. Deshpande
Oriental Pharmacy and Experimental Medicine. 2010; 10(2): 134
[VIEW] | [DOI]
9 Effect of Achyranthes aspera (Linn) seeds on redox and oxidative status in plasma and selected tissues of rats fed with high doses of fructose
Malarvili, T., Gomathi, N.
Biosciences Biotechnology Research Asia. 2009; 6(2): 659-664
10 Studies on antioxidant and antiradical activities of Dolichos biflorus seed extract
Hazra, B., Sarkar, R., Mandal, S., Biswas, S., Mandal, N.
African Journal of Biotechnology. 2009; 8(16): 3927-3933
11 The antioxidant activity and free radical-scavenging capacity of dietary phenolic extracts from horse gram (Macrotyloma uniflorum (Lam.) Verdc.) seeds
Siddhuraju, P., Manian, S.
Food Chemistry. 2007; 105(3): 950-958


Print this article  Email this article


Site Map | Home | Contact Us | Feedback | Copyright and Disclaimer | Privacy Notice
Online since 20th July '04
Published by Wolters Kluwer - Medknow