Therapeutic Potential of Bone Marrow-derived Mesenchymal Stem Cells on CCl4-induced Mouse Hepatic Injury Possibly through Matrix Metalloproteases and Differentiation into Hepatocyte-Like Cells

V. Rabani *, M. Shahsavani , M. Gharavi , Z. Azdari , A. Piriai , H. Baharvand ,


Objective: To study the therapeutic effect of mesenchymal stem cells (MSC) on induced liver fibrosis in mice model and to uncover its mechanism Materials and Methods: we infused GFP+ MSCs of male into the tail vein of female mice that received CCl4 injection biweekly to induce liver fibrosis. MSC which were derived from bone marrow obtained from femoral and tibial bones, isolated and proliferated in culture. They were characterized morphologically and by their potential of differentiation to osteo and adipo lineage. Animals were divided into 6 groups: control, CCl4 4 week, CCL4 8 week, CCl4 4week plus MSC, CCl4 8 week plus MSC, CCL4 plus opportunity to regeneration. Liver tissue was examined histopathologically and by software to quantify collagene deposition as the stage of liver fibrosis. Lipid peroxidation were quantified as a marker of liver injury level. Gene expression ratio of the collagen (type I),TIMP1,MMP-9,MMP-13,alph SMA was detected. Immunostaining were done to show homing and differentiation to hepatocyte of the cells and presentation of Hepatic stellate cells(HSC) . liver functions (serum ALT and AST) were estimated for all groups. Results: Quantitative RT-PCR analysis showed administration of MSCs has a significant antifibrotic effect as evidenced by the significant decrease in liver collagen and increase MMP13 gene expression in the CCl4/MSC group compared to the CCl4 group, 4 weeks after transplantation. The expression of αSMA (smooth muscle actin) and TIMP also reduced in CCl4/MSC group. However, this was statistically nonsignificant. Additionally, the expression of MMP9 significant increase in CCl4 treated groups; however, there was no significant change after MSC injection. The reduction in liver collagen confirmed histopathologically by quantitative analysis. Moreover, lipid peroxidation content in transplanted group decreases significantly. Immunostaining of transplanted cells showed GFP-positive cells in the liver and some of them expressed albumin or αSMA. Conclusion: MSCs can enhance recovery of CCl4-injured mouse liver through their influence in reduction collagen deposition by possible effect on matrix metalloptoteases and their capacity to differentiate into hepatocyte-like cells