Interstitial cystitis (IC) is a chronic inflammation of the bladder, accompanied by pain in the bladder, changes in the frequency of urination and discomfort in the pelvic area. The causes of this disease are still unknown, but due to the frequent recurrence of the symptoms of this disease, efforts are being made to discover possible targets for treatment. One possible target is the cannabinoid receptor 2 (CBR2), which is present in the urothelial (epithelial) cells of the mammalian bladder. In my master's thesis, we determined the distribution and the amount of CBR2 in cultures of normal porcine urothelial cells (NPU) and in non-invasive human urothelial papilloma (RT4) cell lines. We used untreated (control) cells and cells treated with tumor necrosis factor α (TNFα), which induces inflammation in cells, to obtain an in vitro model of IC. CBR2 immunofluorescence labeling was used to determine the distribution and amount of CBR2 in cells, and western blotting was used to determine the amount of CBR2 in cells. We found that in untreated NPU cells, CBR2 is present in the cytoplasm, and in NPU cells treated with TNFα, CBR2 is present in the cytoplasm and in the plasmalemma. In untreated as well as TNFα-treated RT4 cells, CBR2 was always present only in the cytoplasm. The amount of receptors increased in NPU cells after induced inflammation and decreased in RT4 cells compared to cells without induced inflammation, but in both cases without statistically significant differences. The distribution pattern of CBR2 was punctate in untreated as well as treated NPU and RT4 cells. The results of my master's thesis indicate differences in the response of healthy urothelial cells and urothelial papilloma cells to TNFα, which was also expressed in differences in the amount and distribution of the expressed receptor. These results thus provide an important basis for further research to understand the role of CBR2 as a therapeutic target for the treatment of IC.
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