In vitro effects of Sutherlandia frutescens water extracts on cell numbers, morphology, cell cycle progression and cell death in a tumorigenic and a non-tumorigenic epithelial breast cell line.
ABSTRACT Sutherlandia frutescens is a South African herb traditionally used for internal cancers, diabetes, a variety of inflammatory conditions and recently to improve the overall health in cancer and HIV/AIDS patients. The in vitro effects of S. frutescens extracts were evaluated on cell numbers, morphology, cell cycle progression and cell death. Dose-dependent studies (2-10 mg/ml) revealed a decrease in malignant cell numbers when compared to their controls. S. frutescens extracts (10 mg/ml) decreased cell growth in a statistically significantly manner to 26% and 49% (P<0.001) in human breast adenocarcinoma (MCF-7) and human non-tumorigenic epithelial mammary gland cells (MCF-12A) respectively after 72 h of exposure. Cell density was significantly compromised and hypercondensed chromatin, cytoplasmic shrinking, membrane blebbing and apoptotic bodies were more pronounced in the MCF-7 cell line. Both S. frutescens-treated cell lines exhibited and increased tendency for acridine orange staining, suggesting increased lysosomal and/or autophagy activity. Flow cytometry showed an increase in the sub G(1) apoptotic fraction and an S phase arrest in both the 5 mg/ml and 10 mg/ml S. frutescens-treated cells. S. frutescens induced an increase in apoptosis in both cell lines as detected by Annexin V and propidium iodide flow cytometric measurement. At 10 mg/ml, late stages of apoptosis were more prominent in MCF-7 S. frutescens-treated cells when compared to the MCF-12A cells. Transmission electron microscopy revealed hallmarks of increased vacuolarization and hypercondensed chromatin, suggesting autophagic and apoptotic processes. The preliminary study demonstrates that S. frutescens water extracts exert a differential action mechanism in non-tumorigenic MCF-12A cells when compared to tumorigenic MCF-7 cells, warranting future studies on this multi-purpose medicinal plant in southern Africa.
- [Show abstract] [Hide abstract]
ABSTRACT: The influence of strigolactones as hormones in plants is not fully characterised even though they are known to affect plant architecture, both above ground and in the roots. Using an in vitro system, the effects of the synthetic auxins 1-naphthalene acetic acid and indole-3-butyric acid (NAA and IBA) and synthetic strigolactones (GR24 and Nijmegen-1) were tested on microplant development of Sutherlandia frutescens, a leguminous medicinal plant native to South Africa. Considerable phytochemical variation in wild populations has led to the proposal of using micropropagation for this species. This will assist with domestication and provide plants with a more predictable chemistry for the phytopharmaceuticals industry. Nodal explants with an axillary bud were grown on Murashige and Skoog (Plant Physiol 15:473–497, 1962) medium [0.8 % (m/v) agar (pH 5.8), 3 % (m/v) sucrose and 0.1 g/L myo-inositol] supplemented with NAA, IBA, GR24 and Nijmegen-1, either singly or in combination. The amino acid profile and secondary metabolite pool was monitored using LC–MS-profiling. Treatment with NAA promoted mass shoot production, whilst a combination of NAA and Nijmegen-1 also positively influenced the accumulation of amino acids, flavonoids (sutherlandins) and terpenoids (sutherlandiosides) that S. frutescens produces. Since these compounds represent the presumed active compounds in this species and the biomarkers used in quality control assessment of S. frutescens tissues harvested for the pharmaceutical industry, this treatment holds promise for the commercial production of Sutherlandia extracts and herbal medications.Plant Cell Tissue and Organ Culture 06/2014; 117(3):401-409. · 2.61 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: The metabolite profiles of Sutherlandia frutescens populations may vary depending on their geographical location, affecting the quality of plant-based pharmaceutical products generated from this species. This paper aims at using metabolic profiling through liquid chromatography mass spectrometry (LC–MS) to assess the metabolite content of seed pods from populations of the medicinal plant S. frutescens growing in geographically different environments. Terpenoid (retention time: 7.5–9.0 min) and flavonoid (retention time: 15.0–19.0 min) regions of the chromatograms were useful in distinguishing between samples and five distinct clusters were revealed after principal component analysis (PCA). This may assist in tracing the region where plants actually come from and the identification at the subspecies level. To increase the class separation and simplify interpretation, we focused on those populations from the arid Karoo and the coastal Gansbaai area, applying orthogonal partial least squares discriminant analysis (OPLS-DA) to organize these into two clear groups. The presence or absence of sutherlandioside B (SU1) and its derivatives contributed significantly to the separation of the Karoo plants from those from the Gansbaai cluster. Processing procedures of herbal products require standardization, but this becomes challenging when plants do not contain key chemical principles. Extracts from the Gansbaai population had virtually undetectable levels of SU1; consequently products manufactured from farmed plants originating from this region may lack these compounds, which are now proposed to be anti-cancerous. There were several sutherlandioside-type metabolites that distinguished populations from each other. These chemicals may add new knowledge in terms of the broader metabolomic understanding of Sutherlandia populations and their potential pharmacological action. In vitro plants generated as part of a commercialization–conservation strategy had a similar metabolite profile to non-propagated plants. In fact, these plants could be traced to the West Coast populations, further confirming their identity. This study highlighted that SU1 cannot be used as the only quality control marker for Sutherlandia products, since it does not occur in all populations and there is no conclusive evidence that it is the main active ingredient of the plant. The effect of α-naphthalene acetic acid (NAA) at 1 mg l− 1 was tested on in vitro plants. Sutherlandiosides and sutherlandins were detectable in treated plants. Although the treatment had impacts on the growth capacity of plants, SU1 did not accumulate at higher levels in auxin-treated plantlets. The similarity of micropropagated plants to wild plants proved that tissue culture does not have deleterious effects on the chemistry of Sutherlandia plants. Metabolomic approaches using LC–MS are thus an important feature as a diagnostic tool and should be integrated into the herbal product manufacturing process utilizing Sutherlandia and its extracts.South African Journal of Botany 09/2012; 82:33-45. · 1.34 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: The surfactant–cobalt(III) complex, cis-[Co(trien)(4CNP)(DA)](ClO4)3 (trien = triethylenetetramine, 4CNP = 4-cyanopyridine and DA = dodecylamine) was synthesized and characterized by various spectroscopic and physicochemical techniques. The critical micelle concentration (CMC) value of this surfactant–cobalt(III) complex in aqueous solution was obtained from conductance measurements. The conductivity data (at 303, 308, 313, 318 and 323 K) were used for the evaluation of the temperature-dependent CMC and the thermodynamics of micellization (ΔG0m, ΔH0m and ΔS0m). Absorption, fluorescence, cyclic voltammetry, circular dichroism and viscosity experiments have been carried out to study the interaction of the surfactant–cobalt(III) complex with DNA and RNA. The results suggest that the complex can bind to nucleic acids by intercalation via the long aliphatic chain of the complex into the base pairs of DNA/RNA. In the presence of an ionic liquid additive, the binding strength of the surfactant–cobalt(III) complex to the nucleic acids increased. The complex was tested in vitro on HepG2 (human hepatocellular liver carcinoma) tumor cell lines and found to be active.New Journal of Chemistry 12/2013; 38(1). · 3.16 Impact Factor