Antioxidante

Indian Journal of Clinical Biochemistry, 2003, 18 (1) 75.79

ALTERED ANTIOXlDANT ENZYME PROFILE IN WOUND HEALING Shivananda Nayak*, Laxminarayana Udupa~and Saraswati Udupa*
* Department of Biochemistry, Kasturba Medica/College, Manipa/ Department of Pharrnacology, Kasturba Medica/ College, Manipa/, Pin: 576119, Karnataka, India

ABSTRACT
The effects of the alcoholic extract of the flowersof Ixora coccinea were studied on some of the antioxidant enzymes in dead space wounds created in rats. Increases in the tensile strength of the wound and in the level of lysyl oxidase, the crucial enzyme for collagen maturation, were observed indicating a definite prohealing action. In addition, a highly significant increase in the levels of catalase, glutathione peroxidase and glutathionereductase was observed in the drug-treated group. Thus, the gain in tensile strength may be attributed not only to the better cross-linking but also to the antioxidant properties of the drug.

KEY WORDS
Antioxidant; catalase; glutathione peroxidase; glutathione reductase; Lysyl oxidase

INTRODUCTION
The flowers of/xora coccinea L. (Rubiaceae) have been used in traditional Indian systems of medicinefor dysentery, anti-inflammatory action (1), treatment of ulcers and more recently, for anti-tumor activity (2). In an earlier study on the effects of this plant on healing, we had reported that the observed increase in tensile strength of dead space wounds was due to increase in glycosaminoglycan and hydroxyproline contents (3). Very few reports are available (4) on the role of antioxidantenzymes on healing. Since this plant was rich in triterpenoids (attributed to have antioxidant action) we took up this study to investigate whether the prohealing action was due to its antioxidant properties.

removed under pressure to yield a solid residue (yield 12%).

Animals
Wistar rats of either sex, bred in the department of Pharmacology, KMC, weighing around 150-200 g were used. They werehoused individually in standardised environmental conditions, fed with pellet rodent diet and water ad libffum. They were made into two groups (control, test) of 6 animals each.

Wound model
Dead space wound model was used (5,6). Dead space wounds of 500 mm 2 were created by implantation of a polypropylene tube (0.5 cm x 2.5 cm) beneath the dorsal paravertebral lumbar skin. Controls received 2ml of 1% carboxymethyl cellulose orally, while the test group received 2 ml of the drug (200 mg/kg body weight) dissolved in 1% carboxymethyl cellulose through the same route, once daily, for a period of 10 days. On the 1lt" postoperative day, the granuloma tissue formed on the dead space wound was excised. Wet weight of the granuloma tissue was noted. In our earlier study (3) we tried withdifferent doses of the drug, 200 mg of drug per kg body weight was shown to be most effective in the process of wound healing.

MATERIALS AND METHODS Preparation of herbal extract
Ixora coccinea flowers cultivated for ornamental purposes were collected and identified in the department of Botany, Poornaprajna College, Udupi.
The shade dried, powdered flowers (220 mg) were soxlet extracted with 90%ethanol. The solvent was
Author for Correspondence:

Dr. S.LUdupa
Professor, Dept. of Biochemistry Kasturba MedicalCollege Manipal Pin:576119,Kamataka,India Telephone: 571201 Ext.22326 Fax: 91 8252 70062 E-mail: s udupa@yahoo.com

C h r o m a t o g r a p h i c techniques detection of triterpenoids

and

Crude ethanol extract from flowers of Ixora coccinea

Indian Journal of C/inica/Biochemistry, 2003

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Indian Journal of Clinical Biochemistry, 2003, 18 (1) 75-79

was used for phytochemical screening. Crude ethanol extract, 100 gm (32.5%) was treated with ether, and then the ether extract was washed with acid and then with alkalis. Only 0.7% by weight of the crude extract of the acid dissolved and alkalidissolved substances were obtained. Ether was completely evaporated...