Evaluation of Antimutagenocity Effect of Lawsonia inermis (henna) Leaf Extract in Swiss albino Mice

 

Wasim Raja1*, RC Agrawal1 and M Ovais2

1Department of Research, Jawaharlal Nehru Cancer Hospital and Research, Centre Idgah hills, BHOPAL (M.P.)

2Department of Biosciences, Barkatullah University Bhopal (M.P.)

* Corresponding Author E-mail: wasimraja_84@yahoo.com

 

ABSTRACT

In the present investigation, antimutagenicity of Lawsonia Inermis leaf extract was studied using a battery of mutagenicity test systems such us micronucleus and chromosomal aberrations in swiss albino mice. This plant is widely used as a cosmetic agent to stain hair, skin and nails. Three groups of Swiss albino mice were selected randomly (group with cyclophosphamide only, group with Lawsonia Inermis leaf extract plus cyclophosphamide and group with Lawsonia Inermis leaf extract only). A Single intraperitonial treatment of different concentrations (i.e. 500, 1000 and 1500 mg/kg body weight) dose of Lawsonia Inermis leaf extract was given along with the well known mutagenic agent to the experimental group. On comparing the outcome of all three groups, experimental and group with Lawsonia Inermis leaf extract showed significant reduction in micronucleus formation and chromosomal aberration in bone marrow cells after 24 hrs of administration. So it can be conclude that Lawsonia Inermis leaf extract can reduce the mutagenic potential of cyclophosphamide.

 

 KEY WORDS Mutagecity, Lawsonia Inermis leaf extract, Bone marrow, micronucleus, chromosomal aberrations           

 

INTRODUCTION:

Lawsonia inermis leaf extract is an herbal plant which grows in abandoned area. This plant is a well known as a cosmetic agent and it is used to stain hair, skin and nails. The alcoholic extract of Lawsonia inermis have reported mild antibacterial activity against Micrococcus glatamicus and E. coli.1 Lawsonia inermis have also reported to have antitubercle activity.2 Leaves of Lawsonia inermis have been used as a prophylactic against dermatological problems. The major phytochemical constituents of Lawsonia inermis have also been found to possess significant anti- inflammatory, analgesics and antipyretic activities.3

 

Wound healing potential of different extracts of Lawsonia inermis leaves was also documented in the rat excision and incision wound models. Recently, this compound has been reported to have the inhibitory growth effect against human colon carcinoma, HCT-16 cells.4 Therefore the present study have been undertaken to assess the antimutagenic effects of Lawsonia inermis leaf extract using micronucleus assay and chromosomal aberrations in Swiss albino mice.

 

MATERIALS AND METHODS:

Lawsonia inermis leaves were collected from local herbal garden of Bhopal and the cyclophosphamide was purchased from Sigma Chemical Co. U.S.A. Swiss albino mice belong 6-8 weeks of age group and 25-30 g body weight were obtained from the animal house and provided standard pellet diet and water ad libitum. Different doses of Lawsonia inermis leaf extract (i.e. 500, 1000 and 1500 mg/kg body weight) was given intraperitonially to mice. The requisite dose of Lawsonia inermis leaf extract was dissolved in double distilled water just before use and administered to 6 animals of each groups. Control mice were injected an equal volume of vehicle alone. The group with cyclophosphamide only received single intraperitonially.

 

TABLE-1 Effect of lawsonia inermis leaf extract on micronucleus formation in mouse bone marrow cells.

Group

MNPCE+

SE

PCE/NCE

RATIO

Cyclophosphamide (50 g/kg)

3.30±0.56

0.455±0.219

Lawsonia inermis leaf extract

 

 

(a) 500 mg/kg

0.666±0.575

1.177±0.482

(b) 1000 mg/kg

0.539±0.447

1.222±0.158

(c) 1500 mg/kg

0.500±0.547

1.271±0.169

Lawsonia inermis leaf extract alone

0.166±0.407

1.191±0.200

 

TABLE- 2 Protection against cyclophosphamide induced chromosomal aberrations by lawsonia inermis leaf extract .

Group

% Chromoaber

Chromatid

break

Frag

Associ

Ring

%Protection

Cyclophosphamide (50 g/kg)

47.80 ±3.16

18.64

3

11

5.08

--

Lawsonia inermis leaf extract

 

 

 

 

 

 

 

(a) 500 mg/kg

28.2 ±0.134

11

5

11

1

41.00

(b) 1000 mg/kg

42.42±0.189

22

--

7

1

11.25

(c) 1500 mg/kg

25.68 ±0.077

7

2

2

4

46.27

Lawsonia inermis leaf extract alone

13.48±0.143

1

2

5

2

71.79

 

The animals were sacrificed at the appropriate time by cervical dislocation and slides of bone marrow micronucleus assay and chromosomal aberration assay, slides were prepared essentially as per modified methods.5-9

 

Micronucleus Assay:

For the micronucleus assay, slides are stained with May Grunwald and Giemsa stain.5-7 Total of 1000 polychromatic erythrocytes (PCE) and nonchromatic erythrocyte (NCE) were scored at a magnification of 1000 X for each animal’s sample The data are expressed as the average number of micronucleated PCE per thousand cells per animal ( + SE) for each group of 6 animals. Micronucleated PCE for the test compound were compared with vehicle control group by one tailed‘t’ test with significance determined at the p<0.05.

 

Chromosomal Aberration Assay:

In case of chromosomal aberration assay, slides were prepared.8,9. A total of 100 plates spread metaphase plates were scored for chromosomal aberrations observed were chromatid break, gap, chromatid fragmentation, polyploidy centromeric association etc. The data expressed as % chromosomal aberration.

 

RESULT:

Number of micronuclei is found decrease with the increase in the concentration of Lawsonia inermis leaf extract (500 mg - 0.666±0.575, 1000 mg - 0.539±0.447 and 1500 mg - 0.500±0.547). On the other hand, there was a very much decrease of micronuclei in case of the group with Lawsonia inermis leaf extract (i.e. 0.285 +11) as compared to the control group (i.e. Group with cyclophosphamide only).

 

In case of chromosomal aberration assay, there was a significant elevation of protection in chromosomal aberrations in group with cyclophosphamide plus Lawsonia inermis leaf extract with the increase in the concentration (500 mg – 41.00, 1000 mg – 11.25 and 1500 mg – 46.27). Very significant finding of the present study is maximum chromosomal protection in group with Lawsonia inermis leaf extract (i.e. 71.79%).

 

DISCUSSION:

The antimutagenicity of Lawsonia inermis leaf extract implies that Lawsonia inermis leaf extract may have potential anticarcinogenic effect on cancer induced by well-known mutagenic agent (I.e. cyclophosphamide). The mechanisms by which Lawsonia inermis leaf extract may reduce the risk of cancer may not solely be via reduction of mutation; but may involve the antioxidant activity, immune systems enhancing properties and high levels of tannins present in Lawsonia inermis leaf extract which help reduce incidence of tumors. Hence, the use of Lawsonia inermis as anticarcinogenic drug and also as an antioxidant may actually be giving protection to human body against mutation of cells and cancer. Also, Lawsonia inermis leaf extract possesses some activity against mutagens which possibly can be enhanced at higher concentration.

 

REFERENCES:

1.     Kirtikar KR and Basu BD. Lythraceae in KR. Kirtikar and BD Basu (eds.) Indian Medicinal plants, International Book Distribtors, Dehradun, India. 1981; 2: 1076- 1080.

2.     Sharma S. Tuberculostatic Activity of Henna (Lawsonia inermis inn.) Ubercle. 1990; 71: 293-295.

3.     Ali AH, Bashir AK and Tanira MO. Anti-inflammatory, antipyretic and analgesic Effect of Lawsonia inermis L. (Henna) in rats. Ind. J. Pharmacology. 1995; 51.

4.     Kamei H, Koide T, Kajima T, Hashimoto Y and Hasegawa M. Inhibition of cell growth in culture by quinones. Cancer Biother. Radiopharm. 1998; 13: 185-188.

5.     Suchmid W. The micronucleus test. Mutation Res. 1975; 31: 9-15.

6.     Aron CS, Sorg R, Zimmer D. The mouse bone marrow micronucleus test: Evaluation of 21 drug candidates. Mutation Res. 1989; 223:129-140.

7.     Agrawal RC and Kumar S. Prevention of Cyclophasphamide induced micronucleus formation in mouse bone marrow in indol-3-carbinol. Food and chemical Toxicology.1998; 36;975-977.

8.     Preston RJ, Dean BJ, Galloway A.F, Mcfee, Sheldy, Mammalian in vivo cytogenetic assay- analysis of chromosomal aberration in bone marrow cells. Mutation Res.1987; 189: 157-165.

9.     Agrawal RC and Kumar S. Prevention of Cyclophasphamide induced chromosomal aberrations in mouse bone marrow in indol-3-carinol.Toxicology Letters. 1999; 106: 137- 141.

 

Received on 26.06.2008    Modified on 02.07.2008

Accepted on 22.07.2008   © RJPT All right reserved

Research J. Pharm. and Tech. 1(3): July-Sept. 2008; Page 278-279