The aim of this study is to isolate and evaluate the anticonvulsant components from the leaves of Leonotis leonorus (L) R.aR. and to see if there is any change in activity with the origin of the plant material and I or the season in which plant material is collected. Therefore, in this study, two sites were chosen for collection of plant material and the collection was made in summer and in winter. Chemical, physical and pharmacological methods were used to isolate, identify and to evaluate compounds isolated from the leaves of Leonotis leonorus for anticonvulsant activity.

All six plants studied (Senna petersiana, Terminalia sericea, Cassine transvaalensis, Elephantorrhiza burkei, Rauvolfia caffra and Anredera cordifolia) proved to have considerable antibacterial activity. The water extracts of five of the six plants tested, showed activity against Bacillus pumilis, B. subtilis and Staphylococcus aureus respectively. Water extracts from S. petersiana showed a significant antibacterial activity by inhibiting all Gram-positive and two Gram-negative bacteria. A cytotoxicity assay of three plants (S. petersiana, T. sericea and A. cordifolia) on primary vervet monkey kidney ceelsl showed that A. cordifolia was the least cytotoxic extract with an ID50 value of 1.560 mg/ml. Both S. petersian and T. sericea showed an ID50 value of 0.024 mg/ml. Cytotoxicity as determined in this study does not necessarily mean that the active compound which can be isolated from these plants will also be toxic. Antiviral activity of S. petersiana, T. sericea and A. cordifoli crude extracts were investigated against herpes simplex virus type I at the non-toxic concentrations. Both T. sericea and A. cordifoli extracts showed to be non-active against HSV -I, but S. petersiana showed a 20 % reduction in replication of the virus after the sixth day of the experiment. Because of the sensitivity and instability of compounds in the root extract of S. petersiana, it was very difficult to isolate any pure compound. Bioassay-guided fractionation of the seeds of S. petersiana resulted in the isolation luteolin. Its structure was identified and confirmed through spectroscopic methods including IH, BC, UV, HMBC and HMBQ. An antibacterial assay of luteolin isolated from the seeds of S. petersiana showed activity against Baccilus cereus, B. pumilis, Streptococcus aureus and Staphylococcus areus at the concentration of I mg/ml. In the assay to assess the possible antiviral activity of luteolin against herpes simplex type I virus, 50% of the virus was inactivated at the concentration of 250 μg/ml. The results of this study have shown that it is possibl4e that the extracts studied, can provide humankind with valuable agents of potential use in the treatment of herpes and some bacterial species. / Dissertation (MSc ( Plant Physiology))--University of Pretoria, 2006. / Plant Science / unrestricted

The use of traditional medicine through the use of medicinal plants in Africa and especially in South Africa has long been considered an important characteristic of people's daily lives and socio-cultural heritage. Cotyledon Orbiculata is among the medicinal plants that are used by South African traditional practitioners for the treatment of epilepsy and painful conditions such as corns, warts, toothache, earache, boils and various other ailments. However, the claim of therapeutic successes of medicinal plants by traditional medicine practitioners are hardly subjected to scientific scrutiny. This study therefore, investigated the anti-epileptic property of Cotyledon Orbiculata by studying the effects of the methanol extract of the plant against chemically induced seizures by pentylenetetrazole, picrotoxin, bicuculline and N-methyl-DL-aspartic acid in mice. The study also investigated the analgestic effects of Cotyledon Orbiculata by studying the effect of the plant extract on pain induced by acetic acid and hot plate thermal stimulation.

The use of traditional medicine through the use of medicinal plants in Africa and especially in South Africa has long been considered an important characteristic of people's daily lives and socio-cultural heritage. Cotyledon Orbiculata is among the medicinal plants that are used by South African traditional practitioners for the treatment of epilepsy and painful conditions such as corns, warts, toothache, earache, boils and various other ailments. However, the claim of therapeutic successes of medicinal plants by traditional medicine practitioners are hardly subjected to scientific scrutiny. This study therefore, investigated the anti-epileptic property of Cotyledon Orbiculata by studying the effects of the methanol extract of the plant against chemically induced seizures by pentylenetetrazole, picrotoxin, bicuculline and N-methyl-DL-aspartic acid in mice. The study also investigated the analgestic effects of Cotyledon Orbiculata by studying the effect of the plant extract on pain induced by acetic acid and hot plate thermal stimulation.

In Africa, use of medicinal plants for the treatment of diabetes is very common. However, efficacy on co-administering of medicinal plants with therapeutic drugs hasn't been fully determined, especially for African medicinal plants. The current study focused on assessing the in vitro modulation effects of three popular African medicinal plants, namely: Aloe ferox, Sutherlandia frutescens and Prunus africana (including five commercial preparations containing these medicinal plants) on two of the most important anti-diabetic drug metabolising enzymes, Cytochrome P450 (CYP450) 2C9 and CYP3A4 and a key drug efflux transporter, P-glycoprotein (P-gp). Vivid® microsome-based screening kits were used to assess inhibitory potency of plants preparations on CYP2C9 and CYP3A4 enzymes activities. The study showed that P. africana was a more potent inhibitor of CYP2C9 and CYP3A4 activity than the corresponding positive controls Ginkgo biloba and St. John's wort, which are known to cause clinically significant drug-herb interactions. S. frutescens leaf extract demonstrated potent to moderate inhibition on both the tested CYP activities, while its commercial products (Promune® and Probetix®) possessed moderate to mild inhibitory effects on the activities of both CYPs. Potent inhibitory effect on CYP2C9 and CYP3A4 was seen with Aloe Ferox®. Prosit® and Aloes powder® showed potent to moderate inhibition on CYP2C9 activity and moderate to mild inhibition on CYP3A4 activity. In addition to CYP450 activity, the present study also investigated the effects of the selected medicinal plant products on the activity of the main drug efflux protein, P-gp. A screening assay was specifically developed to assess the potential for herbal remedies to interact with P-gp mediated drug absorption. The assay is based on the principle of the reversal of drug resistance in modified Caco-2 cells specifically altered to express high iv efflux protein activity. These cells display a multidrug resistance phenotype and the addition of a plant extract containing a P-gp inhibitor or substrate will inhibit or compete with any cytotoxic drug and consequently reverse the drug resistance. The suitability of the assay was confirmed using a known P-gp inhibitor. The study observed that the anti-proliferation effect of vinblastine was significantly enhanced in vinblastine-resistant Caco-2 cells, which have high P-gp expression, when they were exposed to the selected African herbal preparations. This observation indicates that the studied plant preparations may alter P-gp functionality and therefore lead to interference with the absorption of co-administered drugs. The outcomes of this study provide useful information on whether there are any potential drug-herb interactions between the commonly used African medicinal plants and oral anti-diabetic drugs, at the level of CYP and P-gp drug metabolism and could contribute to better therapeutic management of Type II diabetics. However these predicted interactions will need to be verified in a clinical setting.

Commercial marrubiin, aqueous and organic extracts of Leonotis leonurus were tested in vitro for their anticoagulant and antiplatelet activities. The aqueous extract inhibited platelet aggregation by 69.5 percent (100 μg/mL), while the organic extract (100 μg/mL) and marrubiin (5 μg/mL) showed 92.5 percent and 91.6 percent inhibition, respectively, by inhibiting the binding of fibrinogen to glycoprotein IIb/IIIa receptor in a concentration dependent manner. The extracts significantly prolonged activated partial thromboplastin time compared to untreated plasma controls. Fibrin and D-Dimer formation were drastically decreased. The extracts and marrubiin concentration-dependently inhibited calcium mobilization induced by collagen and thrombin. The formation of thromboxane A2 was also significantly reduced by both the extracts and marrubiin. Protein secretion and platelet adhesion were significantly reduced by both the extracts and marrubiin. The organic extract and marrubiin showed a more pronounced effect than the aqueous extracts in all the in vitro assays. The ex-vivo animal model confirmed the results obtained in vitro. Similar to the in vitro studies, activated partial thromboplastin time clotting time was prolonged by marrubiin and the number of aggregated platelets were significantly reduced relative to aspirin. The findings reflect that marrubiin largely contributes to the organic extract's anticoagulant and antiplatelet effect in vitro. INS-1 cells were cultured under normo- and hyperglycaemic conditions. Marrubiin and the two Leonotis leonurus extracts were screened for anti-diabetic activity in vitro. The stimulatory index of INS-1 cells cultured under hyperglycaemic conditions was significantly increased by 60 percent and 61 percent (p<0.01; n=5) in cells exposed to the organic extract (10 μg/mL) and marrubiin (500 ng/mL), respectively, relative to the normoglycaemic conditions. The gene expression of insulin was significantly increased by 76.5 and 71 percent, and of glucose transporter-2 by 93 and 92.5 percent for marrubiin and the organic extract, respectively, under the same conditions stipulated above (p<0.01; n=4). The extract and marrubiin similarly showed an increase in respiratory rate under hyperglycaemic conditions. Marrubiin increased insulin secretion, HDL-cholesterol, while it decreased total cholesterol, LDL-cholesterol and the atherogenic index in the in vivo rat model.

Over the past 25 years, there has been a resurgence of worldwide scientific research in the fields of ethnopharmacology. The Western world has acknowledged the continued use of traditional medicines by the majority of third world countries, and the need for novel drug development. Hence, much of the pharmaceutical research in recent years has focused on the ethnobotanical approach to drug discovery (Light et al., 2005). In South Africa, as in most developing parts of the world, traditional herbal medicine still forms the backbone of rural healthcare. The government health services in South Africa provide only western medical care although the majority of the population consult traditional healers for some or all of their healthcare needs (McGaw et al., 2005). Medicinal plants like Harpephyllum caffrum are used as blood purifiers or emetics (Watt and Breyer-Brandwijk, 1962), and also for treating acne and eczema. The antimicrobial activity of this plant can be used to treat septicaemia, which is ranked the sixth leading cause of death among neonates and the eighth leading cause of death for infants through the first year of life (Heron, 2007). In this study, the plants investigated for antimicrobial activity were Harpephyllum caffrum, Hermannia cuneifolia, Chironia baccifera, Rhigozum obovatum, Felicia muricata and Pentzia incana. These plants were tested against ATTC (American Type Culture Collection) strains and microorganisms isolated from clinical isolates of patients suffering from septicaemia. The assay methods used included the agar diffusion method using the Mast multipoint inoculator, the microtitre dilution method were used to determine the minimum inhibitory concentration, thin layer chromatography fingerprints accompanied by bioautographic assay were used to detect the inhibition of bacterial growth by active compounds separated from plant extracts and the Ames test was required to assess the possibility of bacterial mutagenesis upon the exposure to plant extracts which can lead to carcinogenicity. In agar diffusion method, extracts of Harpephyllum caffrum inhibited nine strains of Candida albicans, three species of Acinetobacter and four strains of E.faecalis. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus and three strains of Staphylococcus aureus. Extracts of Chironia baccifera inhibited one strain of Acinetobacter and five strains of E.faecalis. Extracts of plants Rhigozum obovatum, Felicia muricata, and Pentzia incana showed no antimicrobial activity. In the microtitre dilution method used to determine the minimum inhibitory concentration (MIC), the results were different from the agar diffusion method. More activity was observed. Extracts of Harpephyllum caffrum inhibited three strains of E.coli, six strains of S.aureus, three species of Acinetobacter and one strain of Klebsiella pneumonia. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus, three strains of S.aureus, two strains of K.oxytoca and one species of Acinetobacter. Extracts of Chironia baccifera inhibited three strains of S.aureus, one strain of MRSA, one species of Acinetobacter and one strain of S.haemolyticus. The MIC values ranged from 0.049 to 6.25mg/ml. Using the thin layer chromatography fingerprints, bioautography showed the presence of various inhibitory chemical compounds. Methanol and acetone extracts of Harpephyllum caffrum, separated very well and showed various inhibition zones on exposure to Candida albicans, Enterococcus faecalis and Staphylococcus aureus. The different inhibition zones were recorded as Rf In the Ames test (Maron and Ames, 1983) the methanol and acetone extracts of Harpephyllum caffrum and Hermannia cuneifolia were negative which means they were devoid of any mutagenic properties. Methanol extracts of Harpephyllum caffrum showed similar results in the Ames assay as reported by Verschaeve and Van Staden (2008). values ranging from 0.25 to 0.95. The zones indicate the different inhibiting chemical compounds present in the plant. Petroleum ether, ethyl acetate, chloroform and formic acid were the solvents used in the assay in the ratio 8:7:5:1, respectively. Establishing the antimicrobial activity of these plants contribute to the systematic scientific investigation of indigenous South African medicinal plants.

The worldwide AIDS epidemic is known to have had a profoundly negative social, economic and personal impact and has taken a heavy toll on existing health care systems, particularly in developing countries. South Africa is experiencing an HIV epidemic with enormous social and economic consequences. Lopinavir/ritonavir antiretroviral treatment has been accredited with having a significantly positive effect and is a key advance in controlling HIV morbidity and mortality. An indigenous South African medicinal plant, Sutherlandia frutescens, known for its anti-diabetic properties and immune-boosting effects, is used for treating HIV positive patients suffering from opportunistic infections. Despite the use of the medicinal plant extract as homeotherapeutic medication, there is little evidence of toxicity testing that identifies its potential for interaction with antiretroviral drugs. However, scientific data relating to the mechanism through which Sutherlandia frutescens acts on the immune system has not been comprehensively documented. The aim of this study was to investigate lopinavir/ritonavir induced metabolic abnormalities in rats and whether the introduction of a plant extract of Sutherlandia frutescens would counteract the side effects of ARV medication. The results indicated that the rodents did not become insulin resistant, however, biochemical analysis indicated that extended ARV drug treatment would have caused insulin resistance. Significant morphological changes were found in the livers, kidneys and pancreases of rats exposed to the lopinavir/ritonavir. Rats exposed to the Sutherlandia frutescens plant extract showed improved histopathology with minimal abnormalities.

Thesis (MSc)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: The majority of South Africans still depend on the use of traditional remedies, as these are sometimes the only types of health care systems available, especially within rural communities. South Africa comprises approximately 400 000 traditional healers and an estimated 60 to 80% of individuals consulting such traditional healers. As a result, the over-harvesting of many traditional medicinal plants has become a threat to the country’s species diversity and has resulted in the scarcity of certain medicinal plant species. The non-sustainable use of traditional medicinal plants stems from their intense harvesting from the wild to supply the high demands from urban and rural markets. As a result of the escalating population growth rate; high rural unemployment; and fundamental value attached to traditional medicinal plants (socio-economic factors), the national and regional trade of traditional medicines is currently higher than it has ever been. Another reason for the increased threat to traditional medicinal plants is the degradation and weakening of customary laws that have previously regulated such resources. This study focuses on the use of traditional medicinal plants by the Setswana-speaking community for self-medication and as a form of primary health care. Research was conducted in Kimberley, Northern Cape of South Africa and focuses on the issue of the sustainability of medicinal plant use in the area, specifically on use and users as well as the acquisition of material sold by a single trader and harvesting techniques. This is to determine whether harvesting of medicinal plants is a potential threat to plant communities in the area. To address the shortcomings of medicinal anthropology the study also investigates the impacts of relocation and resettlement of various communities in the area, on plant use, methods of collection, the sustainability of the natural resource, as well as the transmission of Setswana indigenous knowledge inter-generationally. most abundant under high disturbances. Certain species reacted positively to disturbance and were most abundant in disturbed habitats. These included Elephantorrhiza elephantina and a Helichrysum sp. To minimise destructive harvesting in the Kimberley area and to ensure the sustainable harvesting of plant material, it is important that local harvesters are educated on proper harvesting techniques and that local gatherers are educated on sustainability issues as well as other ecologically fundamental issues. / AFRIKAANSE OPSOMMING: Die meeste Suid-Afrikaners is steeds afhanklik van tradisionele geneesmiddels aangesien dit soms, veral in landelike gemeenskappe, die enigste beskikbare gesondheidsorg is. Suid-Afrika het sowat 400 000 tradisionele geneeshere wat deur ’n geraamde 60% tot 80% van individue geraadpleeg word. As gevolg hiervan hou die oorontginning van talle tradisionele medisinale-planthulpbronne ’n bedreiging vir die land se spesiediversiteit in en het dit reeds tot ’n skaarste aan sekere medisinale plante gelei. Tradisionele medisinale plante word tans nievolhoubaar aangewend aangesien dit op groot skaal in die veld geoes word om in die groot vraag van stedelike en landelike markte te voorsien. As gevolg van die stygende bevolkingsgroeikoers, hoë landelike werkloosheidsyfer en die grondliggende waarde wat aan tradisionele medisinale plante geheg word (sosio-ekonomiese faktore), is die nasionale en streekhandel in tradisionele geneesmiddels tans groter as ooit tevore. Nog ’n rede vir die toenemende bedreiging van tradisionele medisinale plante is die verslapping en versagting van gewoonteregwetgewing wat voorheen sodanige hulpbronne gereguleer het. Hierdie studie fokus op die gebruik van tradisionele medisinale plante deur die Setswanagemeenskap vir selfbehandeling en as ’n vorm van primêre gesondheidsorg. Die navorsing vir die studie is in Kimberley in die Noord-Kaapprovinsie van Suid-Afrika gedoen en fokus op die kwessie van volhoubare medisinale-plantgebruik in die gebied, met bepaalde klem op gebruik en gebruikers, die verkryging van middels wat deur ’n enkele handelaar verkoop word, en oestegnieke. Die doel van die navorsing was om te bepaal of die oes van medisinale plante ’n moontlike bedreiging vir plantgemeenskappe in die gebied inhou. Om die tekortkominge van medisinale antropologie aan te pak, ondersoek die studie ook die uitwerking van die verskuiwing en hervestiging van verskeie gemeenskappe in die gebied op plantgebruik, oesmetodes, die volhoubaarheid van die natuurlike hulpbronne, asook die oordrag van inheemse Setswana-kennis oor geslagte heen. Selfbehandeling en die gebruik van tradisionele medisinale plante speel steeds ’n groot rol in Kimberley, aangesien die meeste van die individue wat aan die navorsing deelgeneem het steeds tradisionele geneesmiddels as deel van hulle kultuur en tradisie gebruik. Daar word in ’n uiteenlopende verskeidenheid plantmateriaal handel gedryf. Hoewel sommige van die middels plaaslik ingesamel word, word die meeste van ander dele van die land, en in party gevalle van buurlande soos Lesotho en Swaziland, ingevoer. Hoewel die meeste van die materiaal dus nie plaaslik ingesamel word en dus nie bepaald op hierdie studie betrekking het nie, is dit steeds aanduidend van oes- en volhoubaarheidskwessies in ander dele van die land. Die kruiekenner dryf in sewentig tradisionele medisinale-plantsoorte handel, waarvan party beskermd en erg bedreig is, waaronder Prunus africana en Warburgia salutaris wat slegs in beskermde gebiede in die land voorkom. Prunus africana is ’n gelyste spesie in CITES, aanhangsel 2. Ander bedreigde spesies sluit Ocotea bullata, Bersama lucens, Curtisia dentata en ’n Eugenia-spesie in. Die meeste van die plante wat (in Kimberley) geoes word, is in die vorm van ondergrondse bergingsorgane (uitlopers en bolle). Hoewel hierdie plante van stingelskade en die skade aan ondergrondse bergingsorgane kan herstel, vat hulle swak pos indien hulle oorgebruik en oorontgin word, en kan hulle dus mettertyd al hoe minder voorkom. In hierdie studie word die mettertydse afname in plantbevolkings deur die toename in reisafstande na insamelingspunte aangetoon. Hierdie tendens is egter nie in die handelsprys en -materiaalhoeveelhede oor die afgelope eeu weerspieël nie. Die meeste van die studiedeelnemers het bevestig dat die prys en hoeveelheid van die handelsmateriaal deurentyd betreklik stabiel gebly het. Van die teikenspesies wat vir kwesbaarheid of sensitiwiteit vir ontwrigting ondersoek is, het Withania somnifera, Boophane disticha, Dicoma anomala en Bulbine natalensis die laagste oorlewingspotensiaal en die hoogste ontwrigtingsensitiwiteit getoon. Die meeste van hierdie spesies het in baie klein hoeveelhede op die gekose terreine voorgekom. In die geval van Withania somnifera kon die negatiewe resultate egter met die laereënvalseisoen gedurende daardie betrokke jaar in verband gebring word. Hierdie spesie word oor die algemeen as ’n onkruid in ontwrigte gebiede geklassifiseer en kom meestal onder erg ontwrigte toestande voor. Sekere spesies, soos Elephantorrhiza elephantina en ’n Helichrysum-spesie, het positief op ontwrigting gereageer en het volop in ontwrigte habitatte voorgekom. Om vernietigende oestery in die Kimberley-omgewing te minimaliseer en die volhoubare ontginning van planthulpbronne te verseker, is dit belangrik dat plaaslike plukkers in gepaste oestegnieke, en plaaslike insamelaars oor volhoubaarheidskwessies en ander ekologies belangrike sake opgelei word.

M.Sc. (Botany) / A few poorly studied Cape herbal medicinal plants of historic and potential commercial interest which form part of the traditional medicine system of the Cape Region have been selected for this study. These are Centella glabrata L., Olea europaea L. subsp. africana (Mill.), Rafnia amplexicaulis (L.) Thunb. and Tulbaghia alliacea L.f. Centella C. asiatica is mainly used to treat skin conditions and wounds, tuberculosis and coughs and cooked and eaten as a vegetable in South Africa. The treatment of heartburn in the Wild Coast is a recent record. Decoctions of the leaves and stalks of C. glabrata were used for treating diarrhea and dysentery by the early Cape settlers. C. stenophylla or “klipdagga” is used in the treatment of high blood pressure and diabetes in the Still Bay area. This is also a recent new record. The anatomy of the few Centella species studied differed dramatically both in the leaf lamina and the petiole and could be used to differentiate between the species studied. A wider study would be very interesting and could be useful to distinguish between the species. This could have great potential as the ca. 45 species of Centella are often very difficult to distinguish from one another.