The NLRP3 inflammasome is a multiprotein complex that controls caspase-1 activation and the development of the pro-inflammatory cytokines IL-1 and IL-18, as well as pyroptosis. The NLRP3 inflammasome in murine macrophages is normally triggered by pathogen-associated molecular patterns as well as a variety of structurally unrelated stimuli. The exact mechanism of NLRP3 activation by such a diverse set of stimuli is unknown, although many signaling processes have been proposed, including cytosolic efflux and the inflow of certain ions. As a result, numerous studies have suggested that anion channels play a role in NLRP3 inflammasome formation, although no direct evidence of their participation has been found. This thesis project aims to measure the expressions level of CLIC1 before and after LPS treatment. The measurement was done with the help of reference genes. Using qPCR, potential reference genes were tested for their stability and evaluated by GenEx. The study identified GUSB and TBP as the most stable genes. Using the delta delta cq method, data from qPCR were normalized by reference genes. The results from this analysis showed an upregulation in the expression levels of CLIC1. These results showed that CLIC1 an anion channel plays a role in the activation and formation of the NLRP3 inflammasome and other inflammatory disorders such as oxidative stress. The study identified GUSB and TBP as reference genes in LPS stimulated THP-1 macrophages and an upregulation in the expression levels of CLIC1, leading to speculation that LPS stimulation leads to the translocation of CLIC1 to the plasma membrane and suggests the possibility to target CLICs to treat NLRP3-driven diseases.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-20260 |
| Date | January 2021 |
| Creators | Roskvist, Karldiony |
| Publisher | Högskolan i Skövde, Institutionen för biovetenskap |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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