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Faculty of
Veterinary Science
Fakulteit Veeartsenykunde
Lefapha la Diseanse tša Bongakadiruiwa
Introduction
Tuberculosis (TB) is a disease of worldwide occurrence that affects millions of
people globally. Prevalence of resistant strains and comorbidity with other
disease conditions makes tuberculosis a disease of public health concern.
Plants offer potential novel sources of potent antimicrobial agents in the form
of bioactive secondary metabolites (Cowan, 1999). Anti-tubercular activity has
been reported in some plants as well as in lichen species (Jimenez-Arellanes, et
al., 2003; Gupta et al., 2007). Some plants from the Menispermaceae family
are used traditionally to treat cough and TB-related symptoms hence the need
for scientific enquiry into their potential anti-tuberculosis activity. The aim of
this study was to investigate and isolate potential antimycobacterial
compounds from selected species of the Menispermaceae family.
Methodology
§The antimycobacterial and cytotoxic effect of Cissampelos mucronata, and
two other species from the Menispermaceae family was evaluated;
§Antimycobacterial activity was determined against non-pathogenic and
pathogenic Mycobacterium strains using a two-fold serial dilution assay and
thiazolyl blue tetrazolium bromide (MTT) as the growth indicator (Eloff,
1998).
§Cytotoxicity was determined against Vero African monkey kidney cells using
a tetrazolium based colorimetric assay (Mosmann, 1983).
§The acetone extracts of C. mucronata leaves and stem were the most active
against the strains used i.e. Mycobacterium smegmatis, M. fortuitum, M.
aurum and M. bovis.
§Fractionation of the acetone extract C. mucronata (aerial parts), was carried
out using vacuum liquid chromatography with hexane, ethyl acetate,
acetone and methanol:chloroform (3:7)
§Bioassay-guided isolation of the active compounds was performed on the
ethyl acetate fraction using column chromatography.
Results and Discussion
§The ethyl acetate fraction of C. mucronata was most active with minimum
inhibitory concentration (MIC) of 0.015-0.08 mg/ml against all the
mycobacterial strains. The MIC was equal or close to the MIC of the positive
controls (rifampicin and streptomycin) with most of the strains (Table 1).
§The hexane fraction was not toxic to Vero cells (Table 2) compared to
doxorubicin (positive control).
§One compound was isolated and is in the process of being identified
(Figure 1).
1,2 1 1 3 4 1
R. T. Akande , I. Famuyide , S. Nkadimeng , A. Aro , T. Hlokwe , L. J. McGaw
1 2 3
Phytomedicine Programme, Department of Paraclinical Sciences, Faculty of Veterinary Sciences, University of Pretoria, Pretoria, South Africa. Nigeria Atomic Energy Commission, Abuja, Nigeria. Department of Agriculture and Animal
4
Health, College of Agriculture and Environmental Sciences, University of South Africa, Florida, South Africa. Bacteriology Section, Agricultural Research Council—Onderstepoort Veterinary Institute, Onderstepoort, Pretoria, South Africa.
References
Cowan, M.M. 1999. Clinical Microbiology Reviews 12, 564-582.
Eloff, J.N. 1998. Planta Medica 64, 711-713.
Gupta, V.K. et al. 2007. Pharmaceutical Biology 45, 200-204.
Jimenez-Arellanes, A. et al. 2003. Phytotherapy Research 17,
903-908.
Mosmann, T. 1983. Journal of Immunological Methods 65, 55-63.
Table 1: Minimum inhibitory concentration of fractions against pathogenic and non-
pathogenic strains (mg/ml)
Table 2: Cytotoxicity and selectivity index of fractions against Vero cells
Conclusion
The extract and fractions were active against the non-pathogenic and
pathogenic mycobacteria strains in line with the traditional use of the
plants against TB-related symptoms. The fractions were relatively
cytotoxic except for the hexane fraction which had a good selectivity
index against the mycobacteria. Structural elucidation and biological
evaluation of the isolated compound is ongoing.
Figure 1: Thin layer chromatography of the Cissampelos mucronata crude extract (CM),
ethyl acetate fraction (ET), and isolated compound (CPD)
The antimycobacterial and cytotoxic effect of Menispermaceae species
#2303
CM ET cpd