Cytokines, hepatic cell profiling and cell interactions during bone marrow cell therapy for liver fibrosis in cholestatic mice.

Bone marrow cells (BMC) migrate to the injured liver after transplantation, contributing to regeneration through multiple pathways, but mechanisms involved are unclear. This work aimed to study BMC migration, characterize cytokine profile, cell populations and proliferation in mice with liver fibrosis transplanted with GFP+ BMC. Confocal microscopy analysis showed GFP+ BMC near regions expressing HGF and SDF-1 in the fibrotic liver. Impaired liver cell proliferation in fibrotic groups was restored after BMC transplantation. Regarding total cell populations, there was a significant reduction in CD68+ cells and increased Ly6G+ cells in transplanted fibrotic group. BMC contributed to the total populations of CD144, CD11b and Ly6G cells in the fibrotic liver, related to an increment of anti-fibrotic cytokines (IL-10, IL-13, IFN-γ and HGF) and reduction of pro-inflammatory cytokines (IL-17A and IL-6). Therefore, HGF and SDF-1 may represent important chemoattractants for transplanted BMC in the injured liver, where these cells can give rise to populations of extrahepatic macrophages, neutrophils and endothelial progenitor cells that can interact synergistically with other liver cells towards the modulation of an anti-fibrotic cytokine profile promoting the onset of liver regeneration.

O transplante de células mononucleares de medula óssea contribui para o remodelamento da matriz extracelular durante a regeneração do fígado em ratos com fibrose hepática induzida por ligadura do ducto biliar / Bone marrow mononuclear cells transplantation contributes to extracelular matrix remodelling during liver regeneration in rats with hepatic fibrosis induced by bile duct ligation

A fibrose hepática é o resultado de uma lesão crônica, com a ativação de células inflamatórias e fibrogênicas no fígado, as quais levam a um acúmulo excessivo de proteínas de matriz extracelular (MEC). Essas alterações resultam na morte de células do fígado, com desorganização e perda da função do parênquima hepático. A cirrose é o estágio avançado da fibrose, e culmina na falência hepática, uma condição potencialmente fatal cujo único tratamento efetivo é o transplante de fígado, o qual é limitado pela disponibilidade de órgãos. Na busca por terapias alternativas visando a regeneração hepática, o transplante de células mononucleares de medula óssea (CMMO) mostrou resultados benéficos e promissores em modelos animais e alguns protocolos clínicos. Entre essas células, estão as células-tronco hematopoiéticas e mesenquimais, que apresentam potencial regenerativo e modulador da resposta inflamatória. Este estudo pretendeu avançar na compreensão dos mecanismos pelos quais as CMMO podem ajudar na regeneração hepática. Ratos com fibrose hepática induzida por ligadura do ducto biliar (LDB) foram transplantados com CMMO e comparados com ratos com fibrose sem transplante e ratos normais. Parâmetros hepáticos como componentes da MEC (colágeno total, colágenos tipos I e IV, laminina, metaloproteinases de matriz – MMPs), componentes celulares (células fibrogênicas, células de Kupffer e colangiócitos) e enzimas hepáticas foram analisados por microscopia de luz, microscopia confocal, western blotting e espectrofotometria. Os resultados mostraram que o transplante de CMMO contribui para a regeneração hepática de maneira global, (a) diminuindo o acúmulo de colágeno e laminina; (b) aumentando a produção de MMPs que favorecem o remodelamento da MEC, principalmente por células de Kupffer; (c) normalizando a quantidade de colangiócitos e diminuindo a quantidade de células fibrogênicas; e (d) normalizando os níveis sanguíneos das enzimas hepáticas...

Liver fibrosis results from a chronic injury, with the activation of hepatic inflammatory and fibrogenic cells, which lead to an excessive extracellular matrix (MEC) protein accumulation. These alterations result in the death of liver cells, along with disorganization and loss of function of the hepatic parenchyma. Cirrhosis is the advanced stage of fibrosis, and culminates in hepatic failure, a potentially fatal condition whose only effective treatment is liver transplantation, which is limited by organ shortage. Amid the search for alternative therapies aiming hepatic regeneration, bone marrow mononuclear cell (CMMO) transplantation has shown promising and benefic results in animal models and some clinical protocols. Amongst these cells are hematopoietic and mesenchymal stem cells, which present regenerative potential and modulate inflammatory response. This study aimed to add knowledge on the mechanisms by which CMMO may prompt hepatic regeneration. Rats with liver fibrosis induced by bile duct ligation (LDB) were transplanted with CMMO and compared to fibrotic and normal rats without transplantation. Hepatic parameters such as MEC components (total collagen, collagens types I and IV, laminin and matrix metalloproteinases – MMPs), cellular components (fibrogenic and Kupffer cells, cholangiocytes) and liver enzymes were analyzed through confocal, fluorescence and light microscopy, western blotting and spectrophotometry. Results showed that CMMO contribute to liver regeneration globally, (a) reducing collagen and laminin accumulation; (b) increasing MMP production, specially by Kupffer cells, that favor MEC remodeling; (c) normalizing cholangiocyte numbers and reducing fibrogenic cells; and (d) normalizing plasma levels of hepatic enzymes...