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GRX, a liver stem cell line, is not affected by Hepatitis C sofosbuvir and daclatasvir drug treatment in vitro
Hepatitis C is characterized by chronic inflammation in the liver, who often leads to fibrosis, cirrhosis, and death. Upon inflammatory conditions, hepatic stellate cells (HSCs) are known to promote liver fibrosis by changing its profile from quiescent to fibrogenic, where these cells produce great quantities of extracellular matrix which replaces hepatic functional tissue, impairing liver function. Neverthless, recent studies reported HSCs to act as stem cells in the liver tissue, being able to differentiate into hepatocytes and thereby promoting tissue regeneration. In this report, we aimed to evaluate sofosbuvir and daclatasvir treatment on hepatic stellate cells seeking to see if drugs commonly used for the treatment of hepatitis would affect the endogenous liver stem cell.
Objetivos - Metodologia - Resultados - Discussão dos Resultados/Objectives - Methodology - Results - Discussion of Results/Objetivos - Metodología - Resultados - Discusión de los resultados
Objectives: To evaluate whether hepatitis C drugs sofosbuvir and daclatasvir treatment affects hepatic stellate cells normal functions.
Methodology: GRX cells (hepatic stellate cells line) were analyzed for autophagy, cell cycle, cell proliferation, cell viability and oil red staining assays. Cells were treated for 24h and 72h with 230ng/mL and 620ng/mL sofosbuvir and 400ng/mL and 1000ng/mL daclatasvir, respectively, in addition to a group with the highest concentrations of both drugs combined. These drugs concentrations were defined using hepatitis C patient’s serum concentration found in the literature. Cell viability, cell cycle and autophagy were assessed by markers annexin V-FITC, propidium iodide and acridin orange, respectively, and analyzed by flow cytometry. Fibrogenic or quiescent differentiation was evaluated by intracellular lipid droplets staining with Oil Red O after 9 days of treatment, and percentage of stained area was assessed using Image J software. Cell proliferation was evaluated by cumulative population doubling assay for 7 days, and the cells were counted every 72h.
Results and Discussion: We found no difference between treatments in neither of the parameters tested, indicating Hepatitis C sofosbuvir and daclatasvir drug treatment does not affect hepatic stellate cells autophagy, cell viability, cell proliferation or cell cycle, nor does it alter cell differentiation into quiescent or fibrogenic profile. However, we observed a tendency to a slower proliferation rate (1.316 population double score) in the group treated with 620ng/mL sofosbuvir and 1000ng/mL daclatasvir combined while control group population double score was 2.565.
Considerações Finais/Final considerations/Consideraciones finales
Our preliminary results show the hepatic stellate cell line GRX is not affected by sofosbuvir and daclatasvir treatment, thus indicating these drugs do not alter HSCs natural cell profile.
Palavras-chave/Key words/Palabras clave
hepatic stellate cells, hepatitis C, adult stem cells, in vitro, sofosbuvir, daclatasvir
Mesenchymal stem cells/adultas
MICHELE ARAMBURU SERAFINI, Raquel Ayres, Ana Carolina Henzel Raymundo, Eduardo Cremonese Filippi-Chiela, Themis Reverbel da Silveira, Mário Reis Álvares-da-Silva, Fabiany da Costa Gonçalves, Ana Helena da Rosa Paz