Evaluation of anti-inflammatory Properties of Homoeopathic Gel prearations of calendula officinalis, arnica Montana, echinacea angustifolia and hypericum perforatum by Zymography- An in vitro study

Abstract

Context: Inflammation is a physiological process in response to tissue damage resulting from microbial pathogen, infection, chemical irritation and/or wounding.Aims: We investigated the anti-inflammatory effect of four homoeopathic medicaments Calendula officinalis, Arnica Montana, Echinacea angustifolia and Hypericum perforatum by Zymography a process of detection of enzymatic activity on gel electrophoresis. Materials and Methods: An apparatus for gel electrophoresis was used. Sample preparation (10mg in 1ml DMSO) and buffer preparation (225µl of D/W + 25µl of stock solution) was made. The apparatus was assembled. Then casting of polyacrylamide gel was done and it was allowed to polymerize and then the gel electrophoresis was run at least for one and half hours and later removed and was incubated. After electrophoresis the gel was washed with zymogram renaturing buffer (ie 2.5% Triton X-100). Then it was stained with coomassie blue R-250 and later destained with an appropriate destaining solution. Result: Coomasie blue staining of the gel revealed sites of proteolysis as translucent bands on a dark blue background. It showed a percentage of anti-inflammatory activity for Arnica Montana to be 85% for MMP-2 and 80% for MMP-9, for calendula officinalis 80% for MMP-2 and 75% for MMP-9,for echinacea angustifolia 75% for MMP-2 and 68% for MMP-9 and for hypericum perforatum 70% for MMP-2 and 60% for MMP-9. Conclusion: Our findings confirmed the anti-inflammatory potential of all homoeopathic medicaments used.