Bone Marrow Mesenchymal Stem Cell Transplantation Can Induce Markers Of Alternative Kupffer Cells Activation And Improve Mouse Liver Cirrhosis Induced By CCL4

Objective：1. Isolation and identification of bone marrow mesenchymal stem cells in mice.2. To establishm mouse models of hepatic Cirrhosis induced byCCL4, and thendepletion of partof their kupffer cells by clodronate liposome.3. To observe the repairing effect of the four groups of mouses as well as the changs ofkupffer cells phenotypic and function in vivo,after bone marrow mesenchymal stem celltransplantation for three weeks4. Investigate whether the kupffer cells involved in the process of mesenchymal stemcells improve liver cirrhosisMethods:1. bone marrow mesenchymal stem cells were obtained by whole bone marrowadherent method in vitro, then adherent characteristics of bone marrowmesenchymal stem cells as well as morphological characteristics, combined withflow cytometry surface antigen to identify.2. Hepatic fibrosis in mice was induced by intrraperitoneal (i.p.) injection with2mL/kgbody weight of10%CCl4(Sigma) dissolved in olive oil (Sigma),3times a weekfor up to10weeks. 3. Mouse models of hepatic Cirrhosis were intravenously injected with liposomalclodronate (200μl) to deplete KC, Immunohistochemical observation depletionefficiency.4. Using the models of liver injury induced byCCL4, we compared the development ofliver fibrosis in MSCs translation mice and kupffer cells depletion and MSCstranslation mice with of kupffer cells depletion and control group. Blood analysiswas performed to evaluate albumin (ALB)alanine aminotransferase (ALT) andaspartate aminotransferase (AST). Histological analysis of the livers for fibrosiswere performedResults1. MSCs which were obtained by whole bone marrow adherent method, showed a typical longspindle, fibroblast-like,radial or vortex-like arrangement. Flow cytometry confirmed,CD-29, CD105expression rate is more than85%, CD-34, CD-45expression in15%or less2. The cirrhosis model is to establish success,after CCL4induced for10week.It isconfirmed by Liver HE staining and Sirius red staining.3. liposomal clodronate tail vein injection24hours after immunohistochemistryconfirmed excluded Kupffer cell efficiency of90%or more。4. Three week later, the therapeutic effect of the M group was significantly better thanthe other three groups, significantly reduced liver fibrosis, inflammation of the liverand liver cell necrosis, and liver function has also been significantly improved.Eliminate Kupffer cells group L and LM group treatment effects and no significantdifferences between the control group C, and even cirrhosis of the liver and livercell necrosis intensifying. 5. The number of CD68(+) kupffer cell in M group was significantly lower than thecontrol group, while the number of CD206(+)Kupffer cells was significantlyhigher.6. The mice in group M which had been infused with MSCs had increased mRNAexpression of M2associated markers Arg-1,CD206and IL-10compared withmice in group C. The expression of M1associated markers TNF-α,iNOS in themice of group M were significantly higher than group CConclusions1. Transplanted mesenchymal stem cells ameliorate carbon tetrachloride-inducedliver cirrhosis in mouse.2. Liver kupffer cells involved in the process of mesenchymal stem cell therapy forliver cirrhosis.3. MSCs induce changes to macrophage recruitment and promote a wound-healingphenotype that is associated with amelioration of hepatic fibrosis.