Evaluation of plant extracts : artemisia afra and annona muricata for inhibitory activities against mycobacterium tuberculosis and human immunodeficiency virus

Mycobacterium tuberculosis and Human Immuno-Deficiency Virus (HIV) have a high prevalence in South Africa. The development and spread of drug resistant tuberculosis is a serious problem which is exacerbated by tuberculosis (TB) co-infection in HIV patients. Traditional medicinal plants like Annona muricata and Artemisia afra are used for respiratory ailments and antiviral therapies respectively. The aim of this study was to evaluate Annona muricata (ethanolic extract) and Artemisia afra (ethanolic and aqueous extracts) for inhibitory activities against M. tuberculosis and HIV. In vitro bioassays for anti-TB activity included: microplate alamar blue assay (MABA), flow cytometry and ρ-iodonitrotetrazolium chloride assays while anti-HIV activity was determined using an HIV-1 reverse transcriptase colorimetric ELISA kit and an HIV-1 integrase colorimetric immunoassay. Cytotoxicity of plant extracts were assessed by the MTT assay on Chang Liver and HepG2 cells. Potential synergistic effects were determined using the basis of Combination Index. Potential interactions of plant extracts with drug metabolic pathways were evaluated with the Glutathione-S-Transferase assay kit as well as the CYP3A4 assay kit. A. muricata ethanolic extract exhibited anti-TB activity with MIC 125 μg/mL. MABA was shown to be the most sensitive and effective method for the detection of anti-TB activity. Artemisia afra aqueous extract showed HIV-1 reverse transcriptase inhibition exhibiting ˃85 percent inhibition at 1 mg/mL while the ethanolic extracts of A. afra and A. muricata showed inhibition of HIV-1 integrase activity at ˃86.8 percent and ˃88.54 percent respectively at concentrations >0.5 - 4 mg/mL. The aqueous extract of A. afra displayed inhibition of HIV-1 integrase ˃52.16 percent at 0.5 mg/mL increasing to 72.89 percent at 4 mg/ml of the extract. A. muricata was cytotoxic at an IC50 of 30 μg/mL and 77 μg/mL on Chang Liver and HepG2 cells respectively, whilst A. afra aqueous and ethanol extracts were not cytotoxic to both cell lines. The ethanolic extract of A. muricata showed both antagonistic and synergistic properties at various IC values, when used in conjunction with rifampicin. A. afra ethanolic extract interrupted GST activity while aqueous extracts of A. afra and A. muricata had a slight effect. All extracts interrupted CYP3A4 activity, however the ethanolic extracts of A. muricata and A. afra showed greater inhibition than the aqueous extract of A. afra. These extracts should be investigated further as they could be an important source of compounds for treatment of M. tuberculosis and HIV respectively