Topical application of serine proteases from Wrightia tinctoria R. Br. (Apocyanaceae) latex augments healing of experimentally induced excision wound in mice

Yariswamy, M. and Shivaprasad, H. V. and Joshi, V. and Nanjaraj Urs, A. N. and Nataraju, A. and Vishwanath, B. S. (2013) Topical application of serine proteases from Wrightia tinctoria R. Br. (Apocyanaceae) latex augments healing of experimentally induced excision wound in mice. Journal of Ethnopharmacology, 149 (1). pp. 377-383.

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Official URL: http://doi.org/10.1016/j.jep.2013.06.056

Abstract

Ethnopharmacological relevance Wrightia tinctoria R. Br. (Apocyanaceae) is a folk medicinal plant known to have immunomodulatory, anti-inflammatory and antihemorrhagic potential. Wrightia tinctoria latex is used for treatment of various clinical conditions including psoriasis, blisters, mouth ulcers, and extensively for topical application on fresh wounds to promote accelerated healing. Aims of the study To investigate the wound healing potential of Wrightia tinctoria latex proteases using a mouse model. Materials and methods Proteolytic activity of Wrightia tinctoria latex proteases (WTLP) was determined on various substrates (casein, gelatin and collagen (type-I and IV)). The thermal stability and the class of proteases present in WTLP were determined using heat treatment and specific protease inhibitors, respectively. Excision wound model in mice was used to evaluate the healing potential of WTLP application (twice daily, 10 mg/kg). Neosporin, a standard drug, was used for comparison. The progression of healing was monitored using physical (wound contraction), biochemical (collagen content, catalase and MMP activity) and histological examinations. Results WTLP contains thermostable serine proteases, which are completely inhibited by PMSF. WTLP showed strong caseinolytic, gelatinolytic and collagenolytic activity. The excision wound healing rate upon WTLP treatment was significantly higher than (>2-fold) the control group (49% vs. 18%, *p<0.01) on day 3 and throughout the study. PMSF pre-treated and heat denatured WTLP failed to promote wound healing. In addition, serial biochemical analysis of the granulation tissue demonstrated 1.5-fold more (2444±100 vs. 1579±121 μg/100 mg tissue) hydroxyproline content and 5.6-fold higher catalase activity (16.7±1.3 vs. 3±0.3 units/mg) compared to controls. Further, the enhanced collagen content and matrix metalloproteinase activity correlated with wound contraction rate following WTLP and Neosporin treatment. Histological analysis on day 9 confirmed complete epithelialization, re-establishment of skin structure and accelerated wound healing following WTLP treatment. Conclusions The thermostable serine proteases of Wrightia tinctoria latex are directly involved in the wound healing process. Our findings provide a biochemical basis for the role of WTLP in the enhancement of wound healing. The study supports traditional topical application of Wrightia tinctoria latex on fresh wounds to promote accelerated healing.

Item Type: Article
Uncontrolled Keywords: Administration, Animal, animal experiment, animal model, Animals, Apocynaceae, article, Bovine serum albumin, BSA, controlled study, Disease Models, Drug Stability, ECM, EDTA, Enzyme Stability, Ethnopharmacology, Ethylene diamine tetra-acetic acid, Extracellular matrix, female, Female, heat treatment, histopathology, Hot Temperature, IAA, India, Iodo acetic acid, latex, Latex, Latex proteases, male, Male, matrix metalloproteinase, Matrix metalloproteinases, Mice, MMPs, mouse, Mus, neosporin, nonhuman, p-DMAB, Para dimethyl amino benzaldehyde, Penetrating, Phenyl methyl sulphonyl fluoride, PMSF, Protease inhibitors, Protease Inhibitors, protein degradation, SDS-PAGE, Serine proteases, Serine Proteases, serine proteinase, Sodium dodecyl sulphate polyacrylamide gel electrophoresis, thermostability, Topical, topical treatment, wound contraction, Wound excision healing, wound healing, Wound Healing, Wounds, Wrightia tinctoria, Wrightia tinctoria latex proteases, WTLP
Subjects: C Chemical Science > Biochemistry
Divisions: Department of > Biochemistry
Depositing User: Arshiya Kousar
Date Deposited: 19 Nov 2019 05:40
Last Modified: 19 Nov 2019 05:40
URI: http://eprints.uni-mysore.ac.in/id/eprint/9571

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