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dc.contributor.advisorMukwevho, E.
dc.contributor.authorMunansangu, Brian Moono Ssimapulasi
dc.date.accessioned2021-09-13T07:58:29Z
dc.date.available2021-09-13T07:58:29Z
dc.date.issued2018
dc.identifier.urihttp://hdl.handle.net/10394/37377
dc.identifier.urihttps://orcid.org/0000-0002-8919-0925
dc.descriptionMSc (Biology), North-West University, Mafikeng Campusen_US
dc.description.abstractThe incidence and prevalence of type 2 diabetes mellitus (T2D) has nearly quadrupled since 1980 and is increasing at disturbing rate worldwide, particularly in low and middle -income countries. The causes are complex, but the rise is due in part to increase in the number of people who are overweight, increase in obesity, and in a widespread lack of physical activity. Besides environmental factors, the combination of heterogeneous genetic defects also plays an important role in determining susceptibility to T2D. Skeletal muscle and adipose tissue are the major tissues involved with post-prandial peripheral glucose disposal in response to insulin. Insulin resistance reduces the ability to clear glucose from the circulation resulting in hyperinsulinemia and hyperglycaemia with the development of T2D. On the other hand, the currently used biguanides and the thiazolidinediones (TZDs) activate AMPK pathway, but these drugs have a wide range of unwanted side effects to the users. A new class of up to eight (08) thiosemicarbazone-triazole hybrids (labelled 1a-h) have been developed which have electron-donating groups such as methoxy substituent attached in one of the aromatic rings resulting in superior mechanisms which can alleviate the symptoms of T2D. In this dissertation, we refer to the thiosemicarbazone-triazole hybrid as hybrid compound b´ or TZD throughout this work). This study focused on assessing if newly synthesized thiosemicarbazone-hybrid (hybrid compound b´) can stimulate the major enzyme in cellular and whole-body energy homeostasis, AMP-activated protein kinase (AMPK). We further assessed the influence on transcription genes involved in T2D such as NRF-1, MEF2A and GLUT-4 as well as its antioxidant capabilities. Mouse C2C12 myoblasts were differentiated from myocytes to myotubes by serum deprivation. Myocytes were cultured in media containing 90% DMEM, 10% FBS, and 1% antibiotic (containing penicillin, streptomycin and fungizone). Thereafter, media was switched to differentiation media (70% DMEM, 2% FCS, 10% DMSO) and then to media with 0.75 mM of Sodium palmitate and metformin 1 μM, a total volume of 5 μL (10 mg/mL) hybrid compound was used at various intervals i.e. 4h,16h, and 24h respectively. The effect of treatment with compound b´ on cell viability was evaluated by MTT assay and total Messenger ribonucleic acid (mRNA) was extracted using the Purelink RNA mini kit and converted to cDNA using the Superscript Vilo kit. Catalytic AMPKα was silenced through a predesigned PRKAA2 siRNA, while mRNA extracted from the siRNA was processed using Cells to CT™ kit. Gene expression was analysed via qRT-PCR. Western blot was employed for protein analysis while ChIP assay was used to assess protein-DNA interactions of MEF2A to NRF-1 promoter. Lastly, the FRAP and TEAC assays were used to assess the Ferric Reducing Antioxidant power of the hybrid compound. The study showed that the newly synthesized thiosemicarbazone-triazole hybrid increased expression levels of glucose transporter genes i.e. GLUT-4, MEF2a, and NRF-1 in including lipid metabolising genes such as PPARα, on palmitate-induced insulin-resistance in C2C12 myotubes. The up-regulation of these genes is a positive effect in that they could help in alleviating the symptoms for type 2 diabetes. The results showed that the hybrid compound was dependent on AMPK pathway; thus AMPK (α1 and α2), as silencing resulted in down-regulation of genes associated with glucose transport. Furthermore, the thiosemicarbazone-triazole hybrid exhibited antioxidant potential activity that could mitigate the effects of free radicals that are usually present in type 2 diabetes.en_US
dc.language.isoenen_US
dc.publisherNorth-West University (South Africa)en_US
dc.titleThe role of thiazolidinedione on adenosine monophosphate-activated protein kinase: Implications on diabetesen_US
dc.typeThesisen_US
dc.description.thesistypeMastersen_US
dc.contributor.researchID24350095 - Mukwevho, Emmanuel (Supervisor)


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