Indian Journal of Pharmacology 2001; 33:

283-285

SHORT COMMUNICATION
HEMIGRAPHIS COLORATA ON WOUND HEALING

EFFECT OF HEMIGRAPHIS COLORATA (BLUME) H. G. HALLIER LEAF ON WOUND HEALING AND INFLAMMATION IN MICE
A. SUBRAMONIAM, D. A. EVANS, S. RAJASEKHARAN, G. SREEKANDAN NAIR
Division of Ethnomedicine and Ethnopharmacology, Tropical Botanic Garden and Research Institute, Pacha-Palode (P. O), Thiruvananthapuram - 695 562.
Manuscript Received: 4.10.2000 Revised: 16.1.2001 Accepted: 22.1.2001

SUMMARY

Objective: To study the wound healing and anti-inflammatory properties of Hemigraphis colorata (leaf). Methods: The wound healing property of H. colorata leaf paste (topical application) or suspension (p.o., 1g/kg) was studied using excision wound which was inflicted by cutting away 500 mm2 of the skin on the anterio-dorsal side of mouse. The anti- inflammatory activity of the leaf was evaluated using carrageenaninduced paw oedema model in mice. Results: H. colorata leaf paste when applied on the wound promoted wound healing in mice but oral administration was ineffective. The wound contraction and epithelialisation was faster in the leaf paste applied on mice. The leaf suspension or paste was devoid of anti-inflammatory activity. Conclusion: H. colorata leaf paste promotes excision wound healing in mice.

KEY WORDS

Hemigraphis colorata

wound healing

anti-inflammation.

INTRODUCTION One of the exotic plants adopted to our country, Hemigraphis colorata (Blume) H.G. Hallier (Acanthaceae) is an excellent garden plant. This is a tropical showy species grown in front of houses and garden. This is a prostrate herb with rooting branches, opposite broad cordate and toothed leaves and terminal heads of small white flowers1. The leaves are 6 to 10 cm long and shimmering silvery violet underneath red purple1. It is claimed in folk medicine that the plant has very good wound healing activity. The leaves are ground into a paste and applied on fresh cut wounds. However, pharmacological studies were not carried out on this plant. In the present study the excision wound healing, and anti inflammatory properties of the plant leaf suspension/paste were studied in mice. MATERIALS AND METHODS Collection of plant material: Hemigraphis colorata leaves were collected in the months of April and May,
Correspondence: G. Sreekandan Nair

2000 from the garden of Tropical Botanic Garden and Research Institute (TBGRI) where the plant is cultivated. A voucher specimen of the plant is kept in the herbarium of the institute. Preparation of leaf suspension and paste: The fresh plant leaves were cleaned and ground into a paste and used. A 20% aqueous suspension of the leaves was prepared in 1% gum acacia. This was used for oral administration. The water content of the leaf was found to be about 80%. Animals: Swiss male albino mice (25-30 g) were used. The mice were fed a standard diet and water ad libitum in our animal house facility and maintained under standard laboratory conditions. Excision wound healing2: An excision wound was inflicted by cutting away approximately 500 mm2 full thickness of shaved skin of a predetermined area on the anterio-dorsal side of each mouse. The wounded mice were divided into 4 groups of 6 mice each. They were kept in individual cage. The control group

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A. SUBRAMONIAM et al.

Table 1. Effect of H. colorota leaf on excison wound contraction in mice. Wound area (mm2) Povidoneiodine ointment Oral administration 528 450 346 276 174 150 + + + + + + 22 (100) 17 (85) 15 (65) 9 (52) 8.5 (33) 8 (28) Topical application 523 320 186 80 18 + + + + + 0 18 (100) 13 (61)* 10 (36)** 7.4 (15)** 1.9 (4)**

H. colorata leaf Post wounding days Control Topical application 508 + 368 + 165 + 76 + 10 + 0 20 (100) 14 (72)* 11 (32)** 6.4 (15)** 1.8 (2)**

0 4 8 12 16 18

533 + 18 (100) 460 + 19 (86) 363 + 14 (69) 278 + 14 (52) 172 + 8 (32) 156 + 6 (29)

Values are mean + S. D; n = 6 in each group; * p < 0.05; ** p < 0.01 (compared to control). % wound contraction is given within parentheses

(group I) did not receive any treatment. In one of the experimental groups (groups II), the leaf paste was applied on the wound with a fine brush, daily, till the wound was completely healed. Another group (group III) of mice received daily the fresh leaf suspension (1 g/kg, p.o.). A positive control group (group IV) of mice received topically 5% (w/w) povidone-iodine ointment (Betadine, Win Medicare Ltd.) in an identical manner. Wound contraction rate was monitored by planimetric measurement of the wound area once in 4 days. This was done by tracing the wound surface on a graph paper2. Reduction in the wound area was expressed as % of original wound area. Epithelialisation time was noted as a number of days after wounding required for the scar to fall off leaving no raw wound behind. Acute inflammation: Carrageenan-induced paw oedema in mice was used as a model. 0.12 ml of 1% carrageenan was injected subcutaneously into the right hind paws of mice with the help of a Hamilton microsyringe. After 3 hrs, the mice were killed with an over dose of ether and both the paws were cut identically at the ankle joint and weighed. The difference in weight between the right and left paws gave the amount of oedema developed. In the experimental group fresh leaf paste was applied on the paw just after carrageenan injection. In another group, the

fresh leaf suspension was administered orally (1g/ kg) just after carrageenan administration. 1% diclofenac (Nac gel) was used as a positive control. Two control groups were taken. 1 % gum acacia was administered (p.o.) to one of the control groups for comparison with oral administration of the drug. Statistical analysis: The data were analysed using one-way analysis of variance (ANOVA). Post-hoc comparisons using Dunnetts test were carried out for the analysis. P values <0.05 were considered significant. RESULTS The wound contraction and epithelialisation were faster in H. colorata leaf paste applied mice when compared to control. As shown in Table 1, wound contraction progressed faster when the leaf paste was applied on the wound compared to untreated wounds. The leaf paste adhered to the wound as a covering. In the first two days after wounding, fluid was oozing from the untreated wounds (control) and, to some extent, from povidone-iodine ointment treated wounds. But in the case of H. colorata leaf paste treated wounds, the drug adhered on the wound and prevented the discharges from the wound within a few hrs after the application. In the drug treated mice the wounds were completely healed in less than 18 days whereas in the control animals it

HEMIGRAPHIS COLORATA ON WOUND HEALING Table 2. Effect of H. colorata leaf on carrageenan-induced paw oedema in mice. Group Control Control (1% gum acacia) H. colorata Oral administration (1 g/kg) Topical application Diclofenac Topical application 42 + 5.1* 88 + 7.1 81 + 5.8 Inflammation (mg)# 84 + 6.2 83 + 7.8

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motes cut wound healing in mice. This observation substantiates the use of this plant leaf in folklore medicine in the treatment of wounds. The wound healing mechanism of this plant leaf is not clear. The leaf extracts (1 mg/ml) did not exhibit in vitro antimicrobial activity. Both alcohol and water extracts of this plant leaf were inactive against bacteria (Bacillus subtilis and Pseudomonas aeruginosa) and fungi (Candida albicans and Aspergillus niger) [unpublished observations of the authors]. However, it is possible that the leaf paste by firmly adhearing on the wound may provide a physical barrier to the invading microorganisms. Oral administration of the drug (1 g/kg; wet weight) moderately enhanced immune responses, such as haemagglutination antibody titre against sheep RBC, number of antibody secreting cells in the spleen, blood leucocyte count and peritoneal macrophage number in mice (unpublished observation of the authors). However, the oral administration did not stimulate wound healing. Thus it appears that the moderate immune stimulation is not significantly involved in the wound healing processes. The drug may stimulate angiogenesis (formation of new blood capillaries). In fact our preliminary observations at the time of drug coat falling suggests that it may stimulate angiogenesis. However, this remains to be confirmed. This herb is a promising wound healing promoter worthy of further studies and clinical evaluation.

# Difference in the weights of carrageenan treated and untreated paws. Values are mean + S. D; n = 6 in each group; * p < 0.01 compared to control

took more than 26 days. Even on the 8th day the wound contraction was 68% in the treated mice whereas it was only 31% in the controls. The efficacy of the leaf preparation was comparable to that of povidone-iodine ointment (Table 1). However, in contrast to topical application, oral administration of the leaf suspension, daily at a dose of 1 g/kg (wet weight) did not influence excision wound healing (Table 1). Acute inflammation: As shown in Table 2, both oral administration of the leaf suspension (1g/kg) as well as topical application of the leaf paste were devoid of anti-inflammatory activity in the carrageenan induced acute inflammation in mice. Diclofenac (which was used as a positive control) when applied topically inhibited inflammation by 42% (Table 2). DISCUSSION The present study shows for the first time that application of H. colorata leaf paste on the wound pro-

REFERENCES
1. Graf AB, Exotica international. Vol. 2, New Jercy. Roehrs Company Publishers, 1982. Udupa SL, Udupa AL, Kulkarni DR. Studies on the antiinflammatory and wound healing properties of Moringa oleifera and Aegle marmelos. Fitoterapia 1994;65:11923.

2.

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