An investigation into the antimicrobial and anticancer activities of Geranium incanum, Artemisia afra and Artemisia absinthium

It has been estimated that between 3000 and 4000 plant species are used for their medicinal properties throughout South Africa, with approximately 27 million South Africans making use of traditional medicines. Of this 27 million, 3 million South Africans rely on traditional medicine as their primary source of health care. Of the 250 000 to 500 000 known plant species, very few have been investigated for their pharmacological qualities, and compounds of significant medicinal value may still remain undiscovered in many plant species. The aims of this study included investigating the antimicrobial properties of Geranium incanum and Artemisia afra, both plants traditionally used for their medicinal properties, and comparing the antimicrobial activity of the latter to that of Artemisia absinthium, as well as investigating the anticancer properties of G. incanum and A. afra, and comparing the anticancer activity of the latter to that of A. absinthium. Infusions, aqueous-, methanol- and acetone extracts of the three plants were prepared and used for anticancer and antimicrobial screening. Plant specimens used to prepare extracts for antimicrobial activity were collected and extracted over three seasons, while extracts used for anticancer screening were prepared from plants collected during the summer only. Considerable variation existed in the percentage crude extract yields obtained when different extractants were used, while the season in which the plants were harvested and extracted also appeared to play a significant role in the amount of extract obtained. The plant extracts were screened for antimicrobial activity against various strains of Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus and Bacillus cereus, using an agar dilution method. G. incanum and A. afra possessed activity for C. albicans, while all three plants showed activity for S. aureus and B. cereus. Activity was largely dependent on the extraction method used. iii The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay was used to screen for anticancer activity of the respective extracts, at varying concentrations, against MCF-7 (human breast adenocarcinoma) cells, HT-29 (human colonic adenocarcinoma) cells and HeLa (human cervical cancer) cells. All of the extracts showed cytotoxic activity in all three cell lines to varying extents, depending on the extract used and cell line screened. The acetone extract of A. afra proved to be the most effective inhibitor with the lowest IC50 (2.65 ± 1.05 μg/ml) having been shown in MCF-7 cells. A. afra and A. absinthium showed similar inhibitory patterns, with the methanol- and acetone extracts having been the most potent inhibitors of each of the respective cell lines in general. Fluorescence microscopy employing 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) and propidium iodide (PI) staining indicated that the acetone extract of A. afra induces apoptosis in MCF-7 cells as apposed to necrosis, and the results were comparable to those obtained for cells exposed to cisplatin. Screening of the A. afra acetone extract for toxicity in normal human cells using the CellTiter-Blue® assay indicated the extract to be toxic to peripheral blood mononuclear cells (PBMC’s) at concentrations comparable to that for MCF-7 cells, while cell cycle analysis of MCF-7 cells exposed to the A. afra acetone extract indicated the extract’s ability to induce apoptosis comparable to that of cisplatin, with the extract exerting its activity at a point during or just prior to the S phase of the cell cycle