The in vitro and in vivo anti-flammatory properties and cytotoxicity of extracts of Euphorbia hirta

Asthma is considered one of the most common respiratory complaints in the world today but a medical cure for this condition is currently not available. The use of herbal medicines to treat asthma has however been reported and Euphorbia hirtais one such herb. The alkaloids, flavonoids, glycosides, sterols, tannins and triterpenoids in E. hirta appear to exert the anti-asthma effects reported. In the first part of this study, the aqueous, acetone, dichloromethane and hexane extracts of E. hirta were evaluated for their effects on the lysosomal membrane integrity, cell viability and cell number of MRC-5 cell-line using the NR/MTT/CV assay. Hydrocortisone was used as a pharmaceutical control. The differences between the effects of the different extracts were investigated and the effects of the extracts were compared with hydrocortisone. Results obtained showed that hydrocortisone was relatively toxic to the MRC-5 cells whereas all four extracts studied showed very limited cytotoxic effects, with the aqueous extracts generally exhibiting the least effects. In the second part of this study, the effects of the aqueous E. hirta extract on the blood coagulation system and general airway wall microstructure and ultrastructure were investigated using the BALB/c mouse asthma model. Hydrocortisone was also used as a pharmaceutical control. Parameters studied included inflammatory cell population in peripheral blood and their migration into the lung parenchyma; platelet aggregation and fibrin fibre morphology; fibroblast and mucous cell proliferation; alveolar cell numbers, lamellar body formation as well as filopodia formation. The animal weights were continuously being monitored throughout the study. Results from the animal studies showed that the aqueous extract of E. hirta had limited effects on changes in the animal weights and did not cause fragility of blood fibrin fibres nor change the integrity and morphology of the platelets in the mice as seen in those treated with hydrocortisone. E. hirta extracts also significantly reduced the number of active inflammatory cells (especially neutrophils, eosinophils and basophils); restored the histological alterations observed in respiratory structures studied and had diverse, dose-dependent beneficial ultrastructural effects like reduction of smooth muscle hypertrophy, inhibition of macrophages into the airway parenchyma, among others. The final judgment and conclusion of this study was that the aqueous E. hirta extract did not show cytotoxic effects and could be used for the treatment of asthma in the BALB/c mice at doses ranging 25-62.5mg/kg. Further research leading to clinical trials is recommended after testing the potency of equivalent doses of this extract in other animal asthma models.Thesis (PhD)--University of Pretoria, 2008.AnatomyPhDUnrestricte

Differential sensitivity of two endothelial cell lines to hydrogen peroxide toxicity: Relevance for in vitro studies of the blood–brain barrier

Oxidative stress (OS) has been linked to blood–brain barrier (BBB) dysfunction which in turn has been implicated in the initiation and propagation of some neurological diseases. In this study, we profiled, for the first time, two endothelioma cell lines of mouse brain origin, commonly used as in vitro models of the blood–brain barrier, for their resistance against oxidative stress using viability measures and glutathione contents as markers. OS was induced by exposing cultured cells to varying concentrations of hydrogen peroxide and fluorescence microscopy/spectrometry was used to detect and estimate cellular glutathione contents. A colorimetric viability assay was used to determine changes in the viability of OS-exposed cells. Both the b.End5 and bEnd.3 cell lines investigated showed demonstrable content of glutathione with a statistically insignificant difference in glutathione quantity per unit cell, but with a statistically significant higher capacity for the b.End5 cell line for de novo glutathione synthesis. Furthermore, the b.End5 cells demonstrated greater oxidant buffering capacity to higher concentrations of hydrogen peroxide than the bEnd.3 cells. We concluded that mouse brain endothelial cells, derived from different types of cell lines, differ enormously in their antioxidant characteristics. We hereby recommend caution in making comparisons across BBB models utilizing distinctly different cell lines and require further prerequisites to ensure that in vitro BBB models involving these cell lines are reliable and reproducible

Neuroprotective Activities of Crossyne flava Bulbs and Amaryllidaceae Alkaloids: Implications for Parkinson’s Disease

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases and affects approximately 6.3 million people worldwide. To date, the treatment of PD remains a challenge, as available treatment options are known to be associated with serious side effects; hence, the search for new treatment strategies is critical. Extracts from the Amaryllidaceae plant family as well as their alkaloids have been reported to have neuroprotective potentials. This study, therefore, investigated the biological activities of Crossyne flava and its isolated alkaloids in an in vitro MPP+(1-methyl-4- phenylpyridinium) PD model using SH-SY5Y cells. The effects of the total extract as well as the four compounds isolated from Crossyne flava (i.e., pancratinine B (1), bufanidrine (2), buphanisine (3), and epibuphanisine (4)) were evaluated for cell viability, neuroprotection, levels of reactive oxygen species (ROS), adenosine triphosphate activity (ATP), and caspase 3/7 activity in SH-SY5Y cells. The results obtained showed that pre-treatment with both the extract and the isolated compounds was effective in protecting the SH-SY5Y cells from MPP+ -induced neurotoxicity and inhibited ROS generation, ATP depletion as well as apoptosis induction in the SH-SY5Y cells. The results of this study show that the Amaryllidaceae plant family may be a source of novel compounds for the treatment of neurodegenerative diseases, which validates the reported traditional uses

Reproductive, antioxidant, anti-inflammatory, antimicrobial, protective and antidiabetic activities of Helichrysum Mill. species

South African Helichrysum species are known for diverse medicinal use and treatment of different illnesses. Ethnopharmacological studies have revealed the potential use of Helichrysum plants in drug discovery. Although some of these species have been documented, there is still paucity of information on most species. This review seeks to provide a compilation of documented traditional uses, reproductive potential, antioxidant, anti-inflammatory, antimicrobial, protective and antidiabetic activities and other therapeutic properties of some Helichrysum species of South Africa. Information on Helichrysum cymosum, H. foetidum, H. odoratissimum, H. patulum and H. petiolare were collected from such scientific databases as Google scholar, Scifinder, PubMed, Elsevier, Scopus, Science direct in the form of journal articles, scientific reports, theses and books from the Library of the Cape Peninsula University of Technology. Our findings show that these species have historic values in traditional medicine through their diverse use for the management and treatment of such illnesses as cold, diabetes, headaches, digestive problems, sores and wounds, HIV, cancer etc. Additionally, the reported bioactive constituents isolated from these species have been shown to indicate several activities such as antimicrobial, antioxidant, antidiabetic, neuroprotection, reproductive potentials and others. Thus, the current review highlights the phytochemical and bioactive constituents as well as some of the pharmacological properties of the five selected Helichrysum species with a view to providing validation for their use in the pharmaceutical drug development process

CYTOTOXIC EFFECTS OF FERMENTED AFRICAN LOCUST BEAN SEEDS ON A BREAST CANCER CELL

Objective: There is a growing scientific evidence of the health-enhancing benefits of fermented food especially in cancer treatment and prevention. Fermented African locust beans (FALB) are a condiment with many medicinal activities and consume in many West African countries. Breast cancer is the most common cancer among women globally. This study investigated the cytotoxicity of FALB extracts on breast cancer (MCF-7) cells overtime and at different concentrations. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to quantify the extent of cytotoxicity of the extracts. Results: The aqueous extract of FALB had an half maximal inhibitory concentration (IC50) of 1.51 and 0.98 mg/mL on exposure for 24 and 48 h, respectively, against MCF-7 cells. Comparatively, the IC50 obtained for the same extract against normal human fibroblasts was 1.90 and 1.37 mg/mL, respectively. Conclusion: The results obtained here suggest some measure of selective cytotoxicity by the aqueous extract against transformed as compared with normal cells. These findings present an important lead to the usefulness of this condiment in cancer treatment. Further studies are recommended

Antioxidant and apoptosis-inhibition potential of Carpobrotus edulis in a model of parkinson’s disease

Background: Parkinson’s disease (PD) is a neurological disorder resulting from the progressive loss of dopaminergic neurons. There is currently no known cure for PD, thus the search for complementary and alternative medicines capable of halting the degeneration of dopaminergic neurons is plausible. Carpobrotus edulis (CE) is an indigenous plant used in South African traditional medicine used for the treatment of a number of disease conditions including tuberculosis, diabetes mellitus and constipation. It has been suggested that CE contains bioactive compounds which are responsible for its acclaimed medicinal potential. No studies have been reported on the potential benefit of CE to the nervous system. This study was therefore done to evaluate the protective effects of CE against 1-methyl-4-phenylpyridinium (MPP+)-induced toxicity in the dopaminergic SH-SY5Y cell line, as well as its underlying mechanism. Methods: In this study, SH-SY5Y cells were treated with varying concentrations of CE and MPP+ respectively to determine the optimal concentrations of MPP+ and CE for further experiments. Thereafter, SH-SY5Y cells were pre-treated with 30 μM of CE before exposure to 2 mM of MPP+ to induce cellular damage. Cell viability was evaluated using the MTT assay, intracellular reactive oxygen species (ROS) production was determined using flow cytometry and the Hoechst nuclear staining was used to visualize apoptosis. Caspases 3/7 and 9 activity was assessed using commercially available kits. Results: MPP+ treatment induced marked cell viability, increased the number of condensed nuclei and apoptotic cells, increased ROS production, initiated caspase 9 and activated caspase 3/7 in SH-SY5Y cells. The observed effects of MPP+-induced toxicity were attenuated by the pre-treatment of SH-SY5Y cells with 30 μM of CE. Conclusion: The protective effects of CE against MPP+-induced toxicity in SH-SY5Y cells may be attributed to its antioxidant and anti-apoptotic properties

Neuroprotective activities of crossyne flava bulbs and amaryllidaceae alkaloids: Implications for parkinson’s disease

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases and affects approximately 6.3 million people worldwide. To date, the treatment of PD remains a challenge, as available treatment options are known to be associated with serious side effects; hence, the search for new treatment strategies is critical. Extracts from the Amaryllidaceae plant family as well as their alkaloids have been reported to have neuroprotective potentials. This study, therefore, investigated the biological activities of Crossyne flava and its isolated alkaloids in an in vitro MPP+ (1-methyl-4- phenylpyridinium) PD model using SH-SY5Y cells. The effects of the total extract as well as the four compounds isolated from Crossyne flava (i.e., pancratinine B (1), bufanidrine (2), buphanisine (3), and epibuphanisine (4)) were evaluated for cell viability, neuroprotection, levels of reactive oxygen species (ROS), adenosine triphosphate activity (ATP), and caspase 3/7 activity in SH-SY5Y cells. The results obtained showed that pre-treatment with both the extract and the isolated compounds was effective in protecting the SH-SY5Y cells from MPP+-induced neurotoxicity and inhibited ROS generation, ATP depletion as well as apoptosis induction in the SH-SY5Y cells. The results of this study show that the Amaryllidaceae plant family may be a source of novel compounds for the treatment of neurodegenerative diseases, which validates the reported traditional uses

Cytotoxic and cell cycle arrest properties of two steroidal alkaloids isolated from Holarrhena floribunda (G. Don) T. Durand & Schinz leaves

The plant Holarrhena floribunda (H. floribunda; G. Don) is indigenous to sub-Saharan Africa and is traditionally used to treat several ailments. The present study was carried out to isolate and characterize bioactive compounds with anti-proliferative activity present in H. floribunda extracts. Compounds were isolated from H. floribunda using the bioassay-guided fractionation technique of repeated column chromatography and the step-wise application of the MTT reduction assay to assess antiproliferative bioactivity. The structures of the compounds were identified mainly using NMR. The effects of the isolated compounds on the viability, cell cycle and proliferation of human cancer cell lines (MCF-7, HeLa and HT-29) as well as the non-cancerous human fibroblast cell line (KMST-6) were investigated

Neuroprotective activities of Boophone haemanthoides (amaryllidaceae) extract and its chemical constituents

Parkinson’s disease (PD) is a neurodegenerative condition that progresses as age increases, and some of its major symptoms include tremor and postural and movement-related difficulties. To date, the treatment of PD remains a challenge because available drugs only treat the symptoms of the disease or possess serious side effects. In light of this, new treatment options are needed; hence, this study investigates the neuroprotective effects of an organic Boophone haemanthoides extract (BHE) and its bioactive compounds using an in vitro model of PD involving the toxin 1-methyl-4-phenylpyridinium (MPP+) and SH-SY5Y neuroblastoma cells

In vitro evaluation of the antiproliferative activity of Carpobrotus edulis on human neuroblastoma cells

Neuroblastoma is a solid neuroendocrine tumour located outside the cranial cavity and contributes about 15% of all cancer‑associated deaths in children. Treatment of neuroblastoma is quite challenging and involves the use of chemotherapy, surgery and radiotherapy. Despite treatment strategies, systemic toxicity are setbacks to patient well-being, hence the need for a new and affordable approach. Medicinal plants are of importance in the field of drug discovery for cancer as some notable anti-cancer agents have been isolated from them. In the present study, the anti-cancer activity of aqueous extract of Carpobrotus edulis (C. edulis), a ground-creeping edible medicinal plant was investigated in SK-N-BE(2) and SH-SY5Y neuroblastoma cells. The effect of C. edulis on cell viability and survival was determined using MTT (3-[4,5-dimethylthiazol-2-yl] 2,5 diphenyltetrazolium bromide) and clonogenic assays respectively. Apoptosis was determined using a Caspase-9 assay kit and flow cytometry was used to measure intracellular reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential. The results show that C. edulis inhibits cell viability (IC50 of 0.86 mg/ml and 1.45 mg/ml for SK-N-BE (2) and SHSY5Y cells respectively) and colony formation in the neuroblastoma cells as well as induce apoptosis, which is evidenced by an increase in caspase-9 activity in the cells. C. edulis also led to a loss of mitochondrial membrane potential and increased production of ROS. Collectively, these results suggest that C. edulis induces cell death via induction of mitochondrial-mediated apoptosis and accumulation of intracellular ROS, thus providing a rationale for further investigations