Metilsulfonilmetano atenua a citotoxicidade induzida por vancomicina em linhagem de células de túbulo proximal HK-2, relacionado com a via oxidativa Nrf-2

• Ano de defesa: 2022
• Autor(a) principal: Rodrigues, Rodrigo Alboim de Paiva Fernandes
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Palavras-chave em Português: Injúria Renal Aguda; Vancomicina; Estresse Oxidativo; Células; Rim
• Link de acesso: http://www.repositorio.ufc.br/handle/riufc/69750

The sudden loss of kidney function is called acute kidney injury and even though it is a syndrome, it is traditionally seen as a single disease. Vancomycin (VCM) is a drug commonly used in the clinic, capable of causing acute kidney injury. In the search for treatments for acute injury induced by MCV in vitro, we chose to use the compound Methylsulfonylmethane (MSM) as a possible treatment, given its antioxidant and anti-inflammatory activity described in the literature. We defined the IC50 for MSM in HK-2 cells through the cell viability assay using the MTT method, for 24 hours, with a result of 42.25 mg/mL. We used working concentrations of 12.5 and 25 mg/ml of MSM for the other analyses. Analyzing the concomitant treatment of MSM and 5 mM MCV, we found a significant improvement (p < 0.05) in cell viability at both concentrations when compared to the lesion group with an improvement of 27% and 35% for concentrations of 12.5 mg /mL and 50 mg/mL, respectively. By flow cytometry, we obtained the results, in relation to the control group, of 13.19 % (7AAD¬+/Anx-) related to the cell necrosis mechanism and 18.73 % (7AAD¬-/Anx+) to a mechanism of late apoptosis in the lesion group. MSM at a concentration of 12.5 mg/mL showed an increase in the number of unlabeled cells, with a value of 72.66 % (7AAD¬-/Anx-), while the MSM 25 mg/mL treatment group showed the value of 81.50% (7AAD¬-/Anx-). In the group with MSM 12.5 mg/mL, 15.07% (7AAD¬+/Anx+) was related to the mechanism of late apoptosis and 11.89% (7AAD¬+/Anx-) to cell necrosis. In the MSM 25 mg/mL group, 9.61% (7AAD¬+/Anx+) was related to the mechanism of late apoptosis and 7.16% (7AAD¬+/Anx-) to cell necrosis. The lesion group labeled with DCFH had an IFR value of 3.01, indicating a large production of ROS and the MSM 25 mg/mL group reduced such production with a value of 1.84. The Rho123-labeled lesion group had a value of 0.39, indicating mitochondrial damage. The 25 mg/mL MSM group was able to improve mitochondrial injury with a value of 0.62. The result of molecular docking was the interaction between the Keap1 molecule and the MSM with ΔG of -5.88Kcal/mol and the sharing of an isoleucine residue at position 559 of a hydrogen bond type when compared to the ideal ligand IQK. We observed the occurrence of interaction between the RAF-1 molecule and the MSM with ΔG of -6.26 Kcal/mol. We analyzed the gene expression of the NrF-2, PPAR- O PPAR-Ɣ, SIRT-1 and GSH-red genes. NrF-2 expression increased in relation to the control and lesion groups. PPAR- O PPAR-Ɣ increased in expression in the MSM 25mg/mL group alone and in the treatment group at this concentration. SIRT-1 was reduced in expression in the lesion and positive control groups with MSM 25 mg/mL. GSH-red showed higher expression in the lesion group and in both treatments with 12.5 and 25 mg/mL of MSM. The results of the study demonstrated that MCV, 5 mM for a period of 24 hours is cytotoxic to human renal tubular cells, being responsible for an intense ROS production and mitochondrial damage, causing a reduction in cell viability by increasing apoptosis. MSM improved cell viability in the face of injury, which may be related to the increase in NrF-2 and its interaction with Keap1, which may lead to more than one cell signaling pathway when HK-2 cells are treated with MSM.