| Objective To explore the therapeutic effect of human umbilical cord blood stem cells (UCBSCs) transplantation into a SD rat model of decompensated cirrhosis, based on the determination of liver function, hepatic fibrosis indexes, and pathological score. Thereby, to provide experimental evidences for the application of UCBSCs in the treatment of decompensated cirrhosis.Methods Mononuclear cells were isolated via hydroxythyl starch method, and flow cytometry was adopted to detect umbilical CD34+blood stem cells in the isolated cells. The SD rat model of decompensated cirrhosis was established by peritoneal injection of CCl4-ethanol solution, and was verified with liver function indexes and pathological examination results. The model rats were randomly divided into two groups:the experimental group and the control group,10rats in each group, upon detection of two groups each index differences were not statistically different (P>0.05). The experimental group received UCBSCs via tail vein injection, whereas control group received a single dose of DMEM/F-12, after which each rat was given2mg/100g of intro-peritoneal cyclophosphamide every day for a week. All rats were observed for4weeks after injection, and blood samples were taken for liver function and liver fibrosis index weekly. Survival status and survival time of rats in each group was closely observed and recorded. All survived rats were killed by cervical dislocation four weeks later, and liver tissues were taken for HE and Masson staining for the observation of liver cell injury and cirrhosis analysis, and pathological grading.Results1. Weekly dynamic test demonstrated that, compared with the control group, in the experimental group rats, ALT and AST were decreased significantly in a time-dependent manner (P<0.05), as well as PCIII (P<0.05). The experiment also showed that Alb was slightly increased, whereas TBil, HA, LN, CIV were decreased, the result was not statistically significant (P>0.05).2. Observation of liver specimens:in the control group, livers were hard in texture and dark in color, showing various degree of atrophy and hardened nodules of various sizes on the surface; in contrast, in the experimental group, livers were showing with blunt edges and thickened surface texture. Small particle-like protrusions were observed locally. Nodule formation was significantly reduced compared with the control group.3. HE and Masson staining of liver specimens of the control group showed moderate to severe liver fibrosis, fibrous septa thick pseudo lobule, severe liver cell degeneration and necrosis, large amounts of fat vacuoles, while that of the experimental group showed moderate liver fibrosis, slender interval fiber of pseudo lobule, lesser liver cell degeneration and necrosis, and only small amount of fat vacuoles. Histopathological scores showed that, comparing to the control group, the extent of necrosis and fatty degeneration of liver cells of the experimental group were significantly reduced (P<0.05), however the degree of liver fibrosis reduced was not statistically different(P=0.136).4. After treatment,2rats of the experimental group died, survival rate was80.0%(8/10), while5died in the control group, survival rate was50.0%(5/10). The survival rates of two groups were not significantly different (P=0.259) by the Log-rank test.Conclusions1. MNC with UCBSCs could be isolated from umbilical blood through HES method, and the amount of UCBSCs is sufficient for therapeutic purposes. They are in coincidence with the characteristics of blood stem cell in terms of immunological marks and biological properties, and can be used as a source of hematopoietic stem cell.2. Rat model with chronic liver cirrhosis which using the method of the modified intraperitoneal injection CCl4is high stability and is consistent with the process of the occurrence and development of human liver injury to hepatic fibrosis.3. UCBSCs promote the repair of liver cell injuries and may inhibit the process of liver cirrhosis. |
PDF Full Text Request