Transplantation of Hyaluronic Acid Coated Umbilical Cord Stem Cells Can Reduce Got, Gpt, Liver Fibrosis and Recover Albumin to Normal of Liver Fibrosis Mice and Therapy Mechanism Study

Liver fibrosis (LF) is mainly due to viral hepatitis, autoimmune diseases, alcoholism or chemical drug-induced liver damage caused by a long time. Mesenchymal stem cells (MSC) with immunosuppressive and anti-inflammatory functions, the use of MSC to treat LF are a very feasible. Hyaluronic acid (HA) ligates MSC surface CD44 with anti-inflammatory and immunosuppressive effects. We hypothesize HA should enhance the Wharton’s jelly MSC (WJMSC) in the treatment of LF mice function and effect. Research aims of this project are (1) the use of carbon tetrachloride to establish stable and reliable animal model of LF, transplantation HA-coated WJMSC to treat LF mice, (2) using the ICPL to label of normal mice, the LF mice, stem cells and HA treated mice liver tissue protein, and then nanoLC-MS/MS to analyze the differences between the three groups of protein expression to study the therapy mechanism. Normal mouse serum glutamate oxaloacetate transaminase (GOT) value is 103.67 ± 20.32 U / L (n = 6), glutamate pyruvate transaminase (GPT) value is 34.5 ± 13.91 U / L (n = 6), albumin was 3.0 ± 0.045 g / dL (n = 6). In a dose 1.0 ml / kg of carbon tetrachloride-induced C57BL / 6 mice, weekly injections for two doses after the eight weeks, compares serum GOT value is more significant increase to 679.75 ± 244.27 U / L (n = 8) (p ＜0.05), GPT value is significantly increased to 733.25 ± 389.74 U / L (n = 8) (p ＜0.05), albumin is significantly decreased to 2.72 ± 0.097 g / dL (n = 8) (p ＜0.05) with normal mice. Liver fibrosis area percentage is 1.76 ± 0.34% (n = 8) significantly greater than normal mice 0.18 ± 0.10% (n = 6) (p = 0.012), determined from the above results liver fibrosis can be induced by carbon tetrachloride in mice after eight weeks. We use HA-coated 6 × 106 WJMSC transplanted into LF mice within the liver capsule (stem cells and HA treated mice). After 28 days of treatment, stem cells and HA treated mice serum GOT decreased to 218.83 ± 46.74 U / L (n = 6), GPT decreased to 177.83 ± 50.84 U / L (n = 6), serum albumin is a significant increase to 3.06 ± 0.089 g / dL (n = 6) (P ＜0.05) and slightly higher than normal mice albumin, the liver fibrosis area percentage is 1.54 ± 0.10% (n = 6) significantly less than the sham group 2.10 ± 0.06% (n = 3) (p = 0.009), albumin in the liver tissue is 2.9 times higher than that of LF mice. Therefore, transplantation of HA-coated 6 × 106 WJMSC is indeed effective in treatment of LF mice to liver function recovery and reduce the degree of liver fibrosis. After 28 days of treatment, in stem cells and HA treated mice group liver tissue, there are still a lot of PKH26 staining of WJMSC and MMP9 expression is very large. MMP9 has been reported in the literature can degrade collagen fibers. We will do in situ zymography to confirm a high level MMP9 in the stem cells and HA treated mice group liver tissue with whether gelatinolytic activity. We use the GeLC-ESI-MS/MS to do WJMSC proteomic analysis. Combined data from three WJMSC proteomic analyses, received a total of 4381 protein expression, including WJMSC have MMP and FGF2. FGFR signaling pathway has 61 proteins, identified to the 25 proteins via GeLC-ESI-MS/MS, up 40.98% of the protein involved in FGFR signaling pathway has been identified to us. Recently been reported in the literature of bone marrow mesenchymal stem cells can secrete FGF2 to inhibit liver fibrosis.

Project ID:PB10108-2796
External Project ID:NSC101-2221-E182-057