Pharmacological investigation of some trees used in South African traditional medicine.

Abstract

South Africa is home to a wide diversity of cultural groups, all of which utilize the flora for a variety of purposes. This is true with regard to traditional medicine systems which are similar to those of the rest of Africa south of the Sahara, with diviners (sangomas) and herbalists (inyangas) as the key health providers. In addition, the Country is rich in plant diversity with some 30 000 species of flowering plants - almost one tenth of the worlds recorded higher plants. This incorporates a large diversity of plants including trees, shrubs, herbs, bulbs and corms. The adverse effects of traditional medicinal plants and natural products are not well documented in the literature. Recently, many plants used as food or in traditional medicine have been shown to be potentially mutagenic using in vitro assays. Thus, the scientific evaluation of traditional medicine and medicinal plants is very important to validate claims made on safety and efficiency of such usages. After a survey of the available ethnobotanical literature, ten trees used in South African traditional medicine were selected. These species were: Acacia niolotica subspecies kraussiana, Acacia sieberiana, Albizia adianthifolia, Combretum kraussii, Faidherbia albida, Ficus sur, Prunus africana, Salix mucronata, Terminalia sericea and Trichilia dregeana. Plant parts including leaf, root and bark were collected from each of the selected trees (exceptions were Albizia adianthifolia, Faidherbia albida, Terminalia sericea and Prunus africana) and extracted using ethyl acetate, ethanol and water individually to ensure the extraction of compounds over a wide range of polarities. The extracts (in total, 78) were screened for antibacterial, anti-inflammatory (COX-1 and COX-2) and antiacetylcholinesterase activities and investigated for their potential mutagenic effects using the Ames test. Antibacterial activity was detected using the disc-diffusion and microdilution assays. The extracts were tested against Gram-positive bacteria: Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus and Gram-negative bacteria: Escherichia coli and Klebsiella pneumoniae. Of the 78 different plant extracts 111 tested (final amount of plant material was 1 mg per disc), 84% showed activity against Gram-positive bacteria. From this percentage, 20% also showed activity against Gram-negative bacteria. The best inhibition was observed with ethyl acetate and ethanol root extracts of Terminalia sericea against both Gram-positive and Gram-negative bacteria. In the micro-dilution assay, 55% of the plant extracts showed minimum inhibitory concentration (MIC) values ~ 1.56 mg/ml against Gram-positive and/or Gram-negative bacteria. The ethyl acetate bark extract of Acacia sieberiana and the root and bark ethyl acetate extracts of Acacia nilotica inhibited bacterial growth of both Gram-positive and Gram-negative bacteria at concentrations ~ 0.8 mg/ml. The aqueous leaf extracts of Acacia sieberiana had a low MIC value (0.3 mg/ml) against Gram-negative Kleibsiella pneumoniae and the ethyl acetate extracts of the root inhibited growth of Escherichia coli with an MIC value of 0.1 mg/ml. However, these two extracts showed no activity in the disc-diffusion assay. The MIC values of the neomycin (control) were 0.8 I-Ig/ml and 3.1 I-Ig/ml against Kleibsiella pneumoniae and Escherichia coli respectively. In the anti-inflammatory test, 70% of the plant extracts from different plant parts (leaf, root, bark) of the tree investigated showed strong inhibition in both the CQX-1 and CQX-2 bioassays. The CQX-2 inhibitory effects of aqueous extracts were generally lower when compared to the organic solvent extracts. However, water extracts of Acacia nilotica was an exception (~ 90%). In the acetylcholinesterase inhibitory test, 21% of the plant extracts were active at concentrations ~ 1 mg/ml using the micro-plate assay. The lowest IC50 value was 0.04 mg/ml obtained with an ethanol bark extract of Combretum kraussii. The IC50 value of the galanthamine (positive control) was 2 I-IM. None of the investigated plants showed any potential mutagenic effects with Salmonella typhymurium strain TA 98 using the Ames test. Using bioassay-guided fractionation, anolignan B was isolated from the ethyl acetate root extract of Terminalia sericea. Antibacterial activity of anolignan B was determined using the microdilution assay. The compound possessed activity against both Gram-positive and Gram-negative bacteria. The lowest MIC value (3.8 IJg/ml) was observed with Staphylococcus aureus. MIC value of the neomycin was 1.5 IJg/ml. Anti-inflammatory activity of anolignan B was detected using the CQX-1 and CQX-2 bioasays. The compound showed strong inhibitory activity against CQX-1 and weaker activity against CQX-2. The ICso values were 1.5 mM and 7.5 mM with CQX-1 and CQX-2 respectively. The ICso values of indomethacin were 0.003 mM and 0.186 mM against CQX-1 and CQX-2 respectively. There were no potential mutagenic effects showen by anolignan B against Salmonella typhimurium strain TA 98 in the Ames test. Isolation of anolignan B from Terminalia species and the antibacterial and anti-inflammatory activities observed in this work have not been reported previously and could therefore be recorded as novel biological activities for this compound. These results also support the idea that the use of ethnobotanical data can provide a valuable short cut by indicating plants with specific uses which might likely be sources of biologically active chemicals.