Pharmacological, toxicological and phytochemical evaluation of helichrysum petiolare hilliard & b.l. burtt - an indigenous plant traditionally used in the treatment of diabetes in the eastern cape province of South Africa
- Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
Anti-oxidant and cytotoxicity activity against the HCC70 Breast cancer cell line of aqueous and methanolic extracts of two red algae species from the Eastern Cape Coast, South Africa
- Authors: Basera, Afra Tsitsi
- Date: 2021
- Subjects: Drug resistance in cancer cells , Oxidative stress
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/22589 , vital:52586
- Description: Cancer, despite extensive research, remains one of the leading causes of mortality. Research into plant-derived compounds has gained traction, as some are not cytotoxic towards healthy cells and pose fewer side effects. Marine algae produce secondary metabolites, protecting them from oxidative stress, which causes cardiovascular diseases, cancer, and diabetes. Therefore, this study aimed to evaluate the anticancer potential of selected red algae species, collected from Humewood Beach, Gqeberha in South Africa, against HCC70 (human breast adenocarcinoma) cancer cells. Identification of the species was made through DNA barcoding of the rbcL gene. Methanolic and aqueous extracts of Gelidium pristoides and Laurencia natalensis were evaluated for antioxidant activity, phytochemical content, and anticancer potential. The phytochemical composition was determined using the phenol, flavonoid, flavonol, tannin, and proanthocyanidin content assays. Antioxidant potential was evaluated using the Ferric reducing antioxidant potential, Diphenyl-1- Picrylhydrazyl (DPPH), nitric oxide, 2, 2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity, and total antioxidant capacity (TAC) assays. The resazurin assay was used to determine the antiproliferative and cytotoxic activity of the methanolic extracts and solid-phase extraction fractions against the HCC70 breast cancer cells. The red algae species were identified as G. pristoides and L. natalensis. The aqueous extract of G. pristoides exhibited significantly higher phenols (24.0 mg/g GAE), tannins (31.6 mg/g TAE), and flavonols content (20.5 mg/g RE) than its methanolic extract, or the aqueous and methanolic extracts of L. natalensis. Methanolic extracts of G. pristoides and L. natalensis exhibited significantly higher proanthocyanidin contents (19.4 mg/g and 15.7 mg/g CE) than their respective aqueous extracts (13.0 mg/g and 11.8 mg/g CE). Flavonoids, however, were absent in both species. The aqueous and methanol extracts of Laurencia natalensis and Gelidium pristoides exhibited low inhibitory activities (less than 50%) against DPPH, ABTS, and nitric oxide radicals. Both species also showed low ferric reducing power. All the extracts were antiproliferative and cytotoxic against the HCC70 breast cancer cells, with L. natalensis exhibiting more significant inhibitory activity. However, its Solid Phase Extraction fractions did not display antiproliferative potential, unlike fraction 4 of G. pristoides. The study results suggest that both G. pristoides and L. natalensis possess bioactive compounds with potential anticancer activity. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021
- Authors: Basera, Afra Tsitsi
- Date: 2021
- Subjects: Drug resistance in cancer cells , Oxidative stress
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/22589 , vital:52586
- Description: Cancer, despite extensive research, remains one of the leading causes of mortality. Research into plant-derived compounds has gained traction, as some are not cytotoxic towards healthy cells and pose fewer side effects. Marine algae produce secondary metabolites, protecting them from oxidative stress, which causes cardiovascular diseases, cancer, and diabetes. Therefore, this study aimed to evaluate the anticancer potential of selected red algae species, collected from Humewood Beach, Gqeberha in South Africa, against HCC70 (human breast adenocarcinoma) cancer cells. Identification of the species was made through DNA barcoding of the rbcL gene. Methanolic and aqueous extracts of Gelidium pristoides and Laurencia natalensis were evaluated for antioxidant activity, phytochemical content, and anticancer potential. The phytochemical composition was determined using the phenol, flavonoid, flavonol, tannin, and proanthocyanidin content assays. Antioxidant potential was evaluated using the Ferric reducing antioxidant potential, Diphenyl-1- Picrylhydrazyl (DPPH), nitric oxide, 2, 2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity, and total antioxidant capacity (TAC) assays. The resazurin assay was used to determine the antiproliferative and cytotoxic activity of the methanolic extracts and solid-phase extraction fractions against the HCC70 breast cancer cells. The red algae species were identified as G. pristoides and L. natalensis. The aqueous extract of G. pristoides exhibited significantly higher phenols (24.0 mg/g GAE), tannins (31.6 mg/g TAE), and flavonols content (20.5 mg/g RE) than its methanolic extract, or the aqueous and methanolic extracts of L. natalensis. Methanolic extracts of G. pristoides and L. natalensis exhibited significantly higher proanthocyanidin contents (19.4 mg/g and 15.7 mg/g CE) than their respective aqueous extracts (13.0 mg/g and 11.8 mg/g CE). Flavonoids, however, were absent in both species. The aqueous and methanol extracts of Laurencia natalensis and Gelidium pristoides exhibited low inhibitory activities (less than 50%) against DPPH, ABTS, and nitric oxide radicals. Both species also showed low ferric reducing power. All the extracts were antiproliferative and cytotoxic against the HCC70 breast cancer cells, with L. natalensis exhibiting more significant inhibitory activity. However, its Solid Phase Extraction fractions did not display antiproliferative potential, unlike fraction 4 of G. pristoides. The study results suggest that both G. pristoides and L. natalensis possess bioactive compounds with potential anticancer activity. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021
Optimisation of expression of a rice (Oryza sativa L.cv Nipponbare) plant natriuretic peptide (OsPNP-B) and its functional characterisation
- Authors: Affun, Ogheneochuko Janet
- Date: 2012
- Subjects: Rice , Plant hormones , Water-electrolyte imbalances
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/24354 , vital:62644
- Description: Maintenance of water and solute homeostasis is a key requirement for living systems, and in vertebrates, homeostasis is in part achieved by natriuretic peptides (NP), a family of peptide hormones. A related family of peptide hormones have also been found in the plant kingdom. Plant natriuretic peptides (PNP) are a novel class of plant proteins with two closely related homologous genes (AtPNP-A and AtPNP-B) identified in the model plant Arabidopsis thaliana. AtPNP-A has been extensively studied and evidence obtained points to a role in plant water homeostasis. No research has been conducted on the function of PNP-B proteins. In this study, we focus on the bioinformatic analysis of the PNP-B gene in various plants, as well investigating whether PNP-B plays a role in water homeostasis in rice plants exposed to drought stress. Basic local alignment search tool (BLAST) queries of the ―The Gene Index‖, EST and available plant genome databases revealed the presence of the PNP-B mRNA in rice, oil seed rape, oak, leafy spurge, poplar, sugarcane, pepper, cotton, apple and maize. All the identified genome sequences contained a predicted intron/insert, which was not present in the related mRNA sequences. However, RT-PCR analysis revealed the presence of the 101 bp insert in the 976 bp amplified rice PNP-B (OsPNP-B) cDNA and therefore expression was optimized for the protein encoded by only the exon2 sequence as this contained the predicted active site region of PNP-B. OsPNP-B was shown to be translated to a protein of 14 kDa with a sequence similarity to AtPNP-B (54percent), AtPNP-A (37percent) and CjBAp12 (55percent), suggesting two possible functions for PNP-B viz water homeostasis and/or pathogenesis defence. To determine whether PNP-B is involved in water homeostasis, total protein extracted from 4 weeks old (4 leaves stage) rice plants subjected to drought treatment for a period of 24, 48, 72, 120, 168 and 240hrs were resolved by 17percent SDS-PAGE and analysed by western blot analysis. The PNP-B protein was found to be down-regulated during drought stress, implying that PNP-B may play a role in water homeostasis through the release of water from cells rather than the up-take of water as seen At-PNP-A. PNP-B could therefore also be involved in plant defence mechanisms to pathogens where plants induce desiccation of infected leaves, thereby ridding the plant of the relevant pathogen. , Thesis (MSc) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012
- Authors: Affun, Ogheneochuko Janet
- Date: 2012
- Subjects: Rice , Plant hormones , Water-electrolyte imbalances
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/24354 , vital:62644
- Description: Maintenance of water and solute homeostasis is a key requirement for living systems, and in vertebrates, homeostasis is in part achieved by natriuretic peptides (NP), a family of peptide hormones. A related family of peptide hormones have also been found in the plant kingdom. Plant natriuretic peptides (PNP) are a novel class of plant proteins with two closely related homologous genes (AtPNP-A and AtPNP-B) identified in the model plant Arabidopsis thaliana. AtPNP-A has been extensively studied and evidence obtained points to a role in plant water homeostasis. No research has been conducted on the function of PNP-B proteins. In this study, we focus on the bioinformatic analysis of the PNP-B gene in various plants, as well investigating whether PNP-B plays a role in water homeostasis in rice plants exposed to drought stress. Basic local alignment search tool (BLAST) queries of the ―The Gene Index‖, EST and available plant genome databases revealed the presence of the PNP-B mRNA in rice, oil seed rape, oak, leafy spurge, poplar, sugarcane, pepper, cotton, apple and maize. All the identified genome sequences contained a predicted intron/insert, which was not present in the related mRNA sequences. However, RT-PCR analysis revealed the presence of the 101 bp insert in the 976 bp amplified rice PNP-B (OsPNP-B) cDNA and therefore expression was optimized for the protein encoded by only the exon2 sequence as this contained the predicted active site region of PNP-B. OsPNP-B was shown to be translated to a protein of 14 kDa with a sequence similarity to AtPNP-B (54percent), AtPNP-A (37percent) and CjBAp12 (55percent), suggesting two possible functions for PNP-B viz water homeostasis and/or pathogenesis defence. To determine whether PNP-B is involved in water homeostasis, total protein extracted from 4 weeks old (4 leaves stage) rice plants subjected to drought treatment for a period of 24, 48, 72, 120, 168 and 240hrs were resolved by 17percent SDS-PAGE and analysed by western blot analysis. The PNP-B protein was found to be down-regulated during drought stress, implying that PNP-B may play a role in water homeostasis through the release of water from cells rather than the up-take of water as seen At-PNP-A. PNP-B could therefore also be involved in plant defence mechanisms to pathogens where plants induce desiccation of infected leaves, thereby ridding the plant of the relevant pathogen. , Thesis (MSc) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012
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