| Objective:Autoimmune hepatitis (AIH) may be caused by inducing agent, autoimmune reaction, hereditary susceptibility and immune-regulatory networks disorder. Drugs of its standard treatment is glucocorticoid combine with immunosuppressive agent, but the rate of complete remission was only40%in AIH patients and often accompanied by adrenal cortical functions of hyperthyroidism syndrome after long course of treatment. AIH will develop into liver cirrhosis and need liver transplantation eventually. It is very necessary to find other safe, effective and economical treatments. Researchers find mesenchymal stem cells (MSCs) could play critical part in immunosuppression, hepatocytes differentiation and anti-fibrosis. MSCs have been successfully used in clinical treatment of heart transplantation, graft versus host disease, hepatic failure and so on, with no side effect and lower costs. The aim of this research was to investigate wether MSCs could be applied in the treatment of AIH in mice model, to explore its underlying mechanism and to inverestgate the effect of MSCs on the AIH animal model’s formation.Materials and Methods:1. Establishment of AIH mice model:the syngeneic liver antigen was emulsified with complete Freund’s adjuvant (CFA), and injected into C57BL/6mice’s enterocoelia at day1and day8.4weeks after first injection,6mice were sacrificed to identify whether the model were established.2. Efficacy of MSCs transplation in the treatment of AIH mice model:AIH mice were divided into3groups:group1were treated with1×106MSCs per mice, group2were treated by0.25mg per mice per week dexamethasone (DXM) by enterocoelia injection, and group3was control group treated by phosphate buffered saline. Till5th and9th week, mice were sacrificed to test the liver and glutamic-pyruvic transaminase (ALT) in blood.3. Effect of MSCs on the AIH animal model’s formation:It contains2groups:group I mice were given1×106MSCs injection at the first day then emulsified fluid of liver antigen and CFA at the second day, group II mice were given Phosphate buffer solution (PBS) at the first day, then emulsified fluid of liver antigen and CFA at the second day. After4weeks, all mice were sacrificed to test their liver and blood.4. The possible mechanism of MSCs in the treatment of AIH mice:splenocyte was harvest and proliferate by con A (group A), supernatants fluid of MSCs conbined with con A (group B), liver antigen s-100-1(group C), and supernatants fluid of MSCs conbined with con A s-100-1(group D), then do MTT assay, and measure the optical density (OD) value.Results:1. The animal model of AIH was successfully established. The average Knodell scores were5.50±1.975, with average ALT was122.00±15.476.2. Efficacy of MSCs transplation in the treatment of AIH mice model:(1) In the4th week, the average ALT level of mice was122.00±15.476U/L, In the5th week, the average ALT level of group1,2and3were:101.43±9.144U/L115.00±10.315U/L and118.33±15.161U/L respectively. There was a significant difference among3groups (P<0.05), but no difference between group1and2(P>0.05). In the9th week, the average ALT level of group1,2and3were:84.00±6.557U/L、98.83±15.767U/L and107.50±11.777U/L. There was a significant difference among3groups (P<0.05), also a significant difference between group1and2(P<0.05).(2) The average liver Knodell score in the4th week was5.50±1.975. In the5th week, this score of group1,2and3were:3.00±1.528、3.33±1.366and4.67±1.862respectively. There was no difference among3groups (P>0.05), and no difference between group1and2(P>0.05). In the9th week, this score of group1,2and3were:2.00±1.291、2.67±0.516and4.33±2.066. There was a significant difference among3groups (P<0.05), and no difference between group1and2(P>0.05).3. Effect of MSCs on the AIH animal model’s formation:(1) The ALT lever of group Ⅰ and Ⅱ was:52.17±15.741and122.±15.476U/L respectively, there was a significant difference between group Ⅰ and Ⅱ (P<0.05).(2) The liver Knodell scores of group Ⅰ and Ⅱ were:3.33±0.816and5.50±1.975respectively. There was a significant difference between group Ⅰ and Ⅱ (P<0.05).4. The possible mechanism of MSCs in the treatment of AIH mice: The OD value of group A was0.267±0.167, the OD of group B was0.217±0.128, the OD of group B was significantly lower than group A (P<0.05). The OD of group C was0.165±0.187, the OD of group D was0.082±0.051, the OD of group D was significantly lower than group C (P<0.05).Conclusions:1. Intraperitoneal injection of syngeneic liver antigen emulsified with CFA can successfully establish the AIH mice model.2. MSCs transplantation can reduce serum ALT of AIH mice, and the effect may be better than of DXM, it is suggested that MSCs transplation could be a effective method in the treatment of AIH patient.3. MSCs pre-transplantation can significantly reduce the ALT levels and ease the liver infiltration of inflammatory cells of AIH mice. It is showed that MSCs pre-transplation can prevent AIH.4. MSCs supernatant can significantly inhibit T lymphocyte proliferation which caused by antigen, which can play central role in the treatment of AIH mice by MSCs. |
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