The effects of terpenoids on the expression and function of cytokines and adipokines in pre-adipocytes and differentiated adipocytes
- Authors: Bloom, Carri-Ann
- Date: 2017
- Subjects: Terpenes , Cytokines , Fat cells
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/9208 , vital:26479
- Description: CURRENTLY UNDER EMBARGO UNTIL THE 26/4/2019: Type 2 diabetes is a metabolic disorder characterised by inflammation, insulin resistance and the inability of pancreatic β-cells to secrete enough insulin to produce a physiological effect. Obesity and high levels of triacylglycerol’s are associated with the development of Type 2 diabetes. Adipose tissue is an active endocrine organ that secretes various protein and peptide hormones, known as adipokines, which mediate important metabolic functions. In an insulin resistant and hyperglycaemic state, levels of anti-inflammatory adipokines, adiponectin, are reduced, whereas levels of pro-inflammatory cytokines, interleukin-6 and interleukin-1β, are elevated; this results in a shift from an anti- to a pro-inflammatory state that is accompanied by dysfunction and apoptosis of the pancreatic β-cells. Cannabis sativa L. has been traditionally used as an anti-inflammatory agent in Southern Africa, specifically treating snakebites, fever and malaria. Δ9-tetrahydrocannabinol is the main psychoactive compound derived from C. sativa, whereas the other major cannabinoids, cannabinol and cannabidiol, have shown anti-inflammatory and sedative properties respectively. Marrubiin is a compound derived from the plant Leonotis leonurus L. and has been traditionally used as an anti-inflammatory and anti-diabetic agent. To determine the effects of these compounds in a hyperglycaemic state, pre- and differentiated mouse adipocytes (3T3-L1 cells) were exposed for seven and fourteen days to the following treatments: Δ9-tetrahydrocannabinol, cannabidiol, cannabinol, marrubiin, anandamide (an endogenous endocannabinoid) and cannabis extract, individually and in combination, under normal glucose and hyperglycaemic conditions. Levels of adiponectin, interleukin-6, leptin, tumour-necrosis factor-α and interleukin-1β were quantified using mouse enzyme-linked immunosorbent assay kits and Oil Red O staining was carried out to determine lipid distribution and lipid droplet characteristics. Results indicate that various cannabinoids, in combination, mediate an anti-inflammatory effect by decreasing the expression of various pro-inflammatory cytokines, which may have allowed for a shift from a pro- to an anti-inflammatory state by these compounds, and may also contribute to the reduction of lipid, which may be used as a supplementary option to current diabetic treatment regimes.
- Full Text: false
- Date Issued: 2017
- Authors: Bloom, Carri-Ann
- Date: 2017
- Subjects: Terpenes , Cytokines , Fat cells
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/9208 , vital:26479
- Description: CURRENTLY UNDER EMBARGO UNTIL THE 26/4/2019: Type 2 diabetes is a metabolic disorder characterised by inflammation, insulin resistance and the inability of pancreatic β-cells to secrete enough insulin to produce a physiological effect. Obesity and high levels of triacylglycerol’s are associated with the development of Type 2 diabetes. Adipose tissue is an active endocrine organ that secretes various protein and peptide hormones, known as adipokines, which mediate important metabolic functions. In an insulin resistant and hyperglycaemic state, levels of anti-inflammatory adipokines, adiponectin, are reduced, whereas levels of pro-inflammatory cytokines, interleukin-6 and interleukin-1β, are elevated; this results in a shift from an anti- to a pro-inflammatory state that is accompanied by dysfunction and apoptosis of the pancreatic β-cells. Cannabis sativa L. has been traditionally used as an anti-inflammatory agent in Southern Africa, specifically treating snakebites, fever and malaria. Δ9-tetrahydrocannabinol is the main psychoactive compound derived from C. sativa, whereas the other major cannabinoids, cannabinol and cannabidiol, have shown anti-inflammatory and sedative properties respectively. Marrubiin is a compound derived from the plant Leonotis leonurus L. and has been traditionally used as an anti-inflammatory and anti-diabetic agent. To determine the effects of these compounds in a hyperglycaemic state, pre- and differentiated mouse adipocytes (3T3-L1 cells) were exposed for seven and fourteen days to the following treatments: Δ9-tetrahydrocannabinol, cannabidiol, cannabinol, marrubiin, anandamide (an endogenous endocannabinoid) and cannabis extract, individually and in combination, under normal glucose and hyperglycaemic conditions. Levels of adiponectin, interleukin-6, leptin, tumour-necrosis factor-α and interleukin-1β were quantified using mouse enzyme-linked immunosorbent assay kits and Oil Red O staining was carried out to determine lipid distribution and lipid droplet characteristics. Results indicate that various cannabinoids, in combination, mediate an anti-inflammatory effect by decreasing the expression of various pro-inflammatory cytokines, which may have allowed for a shift from a pro- to an anti-inflammatory state by these compounds, and may also contribute to the reduction of lipid, which may be used as a supplementary option to current diabetic treatment regimes.
- Full Text: false
- Date Issued: 2017
Cytokine properties of CD23 on human Eosinophilic cells
- Authors: Ferreira, Lauren
- Date: 2007
- Subjects: Cytokines , CD23 antigen , Eosinophil
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10333 , http://hdl.handle.net/10948/503 , http://hdl.handle.net/10948/d1012001 , Cytokines , CD23 antigen , Eosinophil
- Description: CD23, the low affinity IgE receptor, is expressed by various cell types and has numerous functions depending on the form of the protein, its interaction with various ligands and the type of cell involved. CD23 is pivotal in the regulation of IgE, with the soluble form involved in up-regulation, while the membrane bound form is involved in the down-regulation. It is clear why it is believed to be a central molecule in allergic responses, and a therapeutic target for the treatment of allergic disease. In this study a recombinant form of the entire extracellular domain of the protein, exCD23, was produced by PCR cloning and expressed in E. coli. His•Tag™s were introduced onto the C-terminus and N-terminus, respectively, in order to simplify the purification procedure. After renaturation and purification, the recombinant exCD23 bound IgE, indicating its activity. From the IgE binding studies it was established that the position of the tag did not influence the binding. GST•Tagged™ exCD23 was also produced in an attempt to increase the solubility of the recombinant protein, but this proved unsuccessful. Butyrate differentiated EoL-1 cells were treated with the Nterminal His•Tagged™ exCD23, and the protein appeared to suppress the secretion of the constitutively expressed cytokines, especially IL-8 and IFN- , when compared to untreated cells. In addition, treatment of the EoL-1 cells with exCD23 had a significant proliferative effect, but could not induce differentiation of this cell line into mature eosinophilic-like cells.
- Full Text:
- Date Issued: 2007
- Authors: Ferreira, Lauren
- Date: 2007
- Subjects: Cytokines , CD23 antigen , Eosinophil
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10333 , http://hdl.handle.net/10948/503 , http://hdl.handle.net/10948/d1012001 , Cytokines , CD23 antigen , Eosinophil
- Description: CD23, the low affinity IgE receptor, is expressed by various cell types and has numerous functions depending on the form of the protein, its interaction with various ligands and the type of cell involved. CD23 is pivotal in the regulation of IgE, with the soluble form involved in up-regulation, while the membrane bound form is involved in the down-regulation. It is clear why it is believed to be a central molecule in allergic responses, and a therapeutic target for the treatment of allergic disease. In this study a recombinant form of the entire extracellular domain of the protein, exCD23, was produced by PCR cloning and expressed in E. coli. His•Tag™s were introduced onto the C-terminus and N-terminus, respectively, in order to simplify the purification procedure. After renaturation and purification, the recombinant exCD23 bound IgE, indicating its activity. From the IgE binding studies it was established that the position of the tag did not influence the binding. GST•Tagged™ exCD23 was also produced in an attempt to increase the solubility of the recombinant protein, but this proved unsuccessful. Butyrate differentiated EoL-1 cells were treated with the Nterminal His•Tagged™ exCD23, and the protein appeared to suppress the secretion of the constitutively expressed cytokines, especially IL-8 and IFN- , when compared to untreated cells. In addition, treatment of the EoL-1 cells with exCD23 had a significant proliferative effect, but could not induce differentiation of this cell line into mature eosinophilic-like cells.
- Full Text:
- Date Issued: 2007
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