Exploring the interactions of hyperglycaemia, SARS-CoV-2 spike protein & iron in HepG2 cells

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dc.contributor.advisorNell, Theo A. en_ZA
dc.contributor.advisorJoseph, Danzilen_ZA
dc.contributor.authorBreytenbach, Jacomi Heidien_ZA
dc.contributor.otherStellenbosch University. Faculty of Medicine and Health Sciences. Dept. of Biomedical Sciences. Division of Medical Physiology.en_ZA
dc.date.accessioned2024-03-06T12:57:25Zen_ZA
dc.date.accessioned2024-04-26T14:07:14Zen_ZA
dc.date.available2024-03-06T12:57:25Zen_ZA
dc.date.available2024-04-26T14:07:14Zen_ZA
dc.date.issued2024-03en_ZA
dc.descriptionThesis (MSc)--Stellenbosch University, 2024.en_ZA
dc.description.abstractENGLISH ABSTRACT: Background: Diabetic COVID-19 patients have increased severity, disease progression and mortality, as well as an increased risk to develop persisting symptoms. Hyperglycaemia could be a central driving force of the pathophysiology in diabetic COVID-19 patients, demonstrating a multifactorial interaction with metabolic abnormalities, immune dysfunction, and dysregulated iron metabolism. An intricate relationship between hypoxia inducible factor 1, iron and cellular metabolism was also uncovered. Considering the need for a greater mechanistic understanding regarding this interplay, an in vitro investigation into the interaction between glucose and SARS-CoV2 Spike protein (protein involved in viral entry) and the effect of this on HIF-1 activation, iron and inflammatory parameters was therefore undertaken. Methods: HepG2 cells were cultured, and the following treatment groups were allocated: Low glucose (LG), High glucose (HG), Vehicle control (DMSO), LG + Spike protein, HG + Spike protein, HG + Prolyl Hydroxylase Inhibitor (PHI) (HIF-1α stabilizer), and HG + PHI + Spike protein. Various assays were used to assess cell viability, and production of reactive oxygen species (ROS). Western Blots were employed to measure iron, inflammatory, metabolic, and viral entry factor expression. Results: Significant decreases in ferritin levels were observed in HG + Spike, and HG + PHI treatment groups, indicating an interesting interaction between glucose and SARS-CoV-2 Spike that could underly attenuated circulating iron in COVID-19 patients. No significant differences in ROS, or inflammatory parameters were noted between different groups, indicating a lack of an inflammatory response, potentially mediated by low levels of intracellular iron. Lastly, stabilization of HIF-1α, as well as protein expression in different groups, could not be determined due to protein fragmentation. Conclusion: A remarkable interaction between glucose and Spike was associated with lower intracellular ferritin levels, independent of inflammation, necessitating more research.en_ZA
dc.description.abstractAFRIKAANSE OPSOMMING: Agtergrond: Diabetiese COVID-19-pasiënte het verhoogde graad van siekteprogressie en verhoogde mortaliteit, asook 'n verhoogde risiko om volgehoue simptome te ontwikkel. Hiperglisemie kan 'n sentrale dryfveer van die patofisiologie by diabetiese COVID-19-pasiënte wees, wat 'n multifaktoriale interaksie met metaboliese abnormaliteite, immuundisfunksie en gedisreguleerde ystermetabolisme toon. 'n Ingewikkelde verband tussen hipoksie-induseerbare faktor 1, yster en sellulêre metabolisme is ook ontbloot. Die behoefte aan meganistiese navorsing wat lig skyn op hierdie wisselwerking, het gelei tot 'n ondersoek na die interaksie tussen glukose en SARS-CoV-2 Spike-proteïen, bemiddel deur HIF-1-aktivering, en die effek hiervan op yster- en inflammatoriese parameters. Metodes: HepG2-selle is gekweek, en die volgende behandelingsgroepe is toegeken: Lae glukose (LG), Hoë glukose (HG), Draer kontrole (DMSO), LG + Spike proteïen, HG + Spike proteïen, HG + Prolielhidroksilase Inhibeerder (PHI) (HIF-1α stabiliseerder), en HG + PHI + Spike proteïen. Verskeie toetse is gebruik om sellewensvatbaarheid en produksie van reaktiewe suurstofspesies (ROS) te bepaal. Westerse blattering is gebruik om yster, inflammatoriese, metaboliese en virale toegangsfaktor-uitdrukking te meet. Resultate: Beduidende afname in ferritienvlakke is waargeneem in HG + Spike, en HG + PHIbehandelingsgroepe, wat dui op 'n interessante interaksie tussen glukose en SARS-CoV-2 Spike wat moontlik onderliggend is aan verswakte sirkulerende yster by COVID-19-pasiënte. Geen beduidende verskille in ROS, of inflammatoriese parameters is tussen verskillende groepe opgemerk nie, wat dui op 'n gebrek aan inflammatoriese respons, wat moontlik bemiddel kan word deur lae vlakke van intrasellulêre yster. Laastens kon stabilisering van HIF-1α, sowel as proteïenuitdrukking in verskillende groepe, nie bepaal word nie as gevolg van proteïenfragmentasie. Gevolgtrekking: 'n Merkwaardige interaksie tussen glukose en Spike is geassosieer met laer intrasellulêre ferritienvlakke, onafhanklik van inflammasie, wat meer navorsing noodsaak.af_ZA
dc.description.versionMastersen_ZA
dc.format.extent114 pagesen_ZA
dc.identifier.urihttps://scholar.sun.ac.za/handle/10019.1/130342en_ZA
dc.language.isoen_ZAen_ZA
dc.language.isoen_ZAen_ZA
dc.publisherStellenbosch : Stellenbosch Universityen_ZA
dc.rights.holderStellenbosch Universityen_ZA
dc.subject.lcshCOVID-19 (Disease) -- Complicationsen_ZA
dc.subject.lcshHyperglycemiaen_ZA
dc.subject.lcshInflammationen_ZA
dc.subject.lcshDiabetesen_ZA
dc.titleExploring the interactions of hyperglycaemia, SARS-CoV-2 spike protein & iron in HepG2 cellsen_ZA
dc.typeThesisen_ZA
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