| background:With the continuous improvement of medical standards, autoimmune hepatitis diagnostic rate on the rise year by year. Autoimmune hepatitis (AIH) is due to the decreased liver immune tolerance, not recognize its liver tissue composition and its immune response, leading to a kind of self-limiting of hepatitis, its style is given priority to with slow onset of disease. The pathogenesis of AIH is unclear, so far the latest idea is that the body, drugs, or infection factor induced by environment, under the action of susceptibility genes, have liver cell membrane itself a target antigen, make the body’s immune function disorder, immune tolerance and the liver itself, by cellular immunity and humoral immune response, leading to autoimmune liver damage. A globulin hematic disease with elevated serum transaminase and high characteristic, autoantibody positive, liver histology change (interface hepatitis, collect abbacy lymphatic plasma cell infiltration) and immunosuppressive therapy response. The onset of the disease in middle-aged and old females, and clinically can see young women patients generally heavier, see a doctor at a high incidence of liver cirrhosis. About50%of patients with AIH onset conceals, about30%of patients diagnosed with liver cirrhosis has appeared,10%～20%decompensated cirrhosis, and even with haematemesis caused by gastric esophagus varicosity burst bleeds black for starting symptoms. At present mainly hormone and immune inhibitor treatment of the disease, and need long-term or lifelong treatment, long-term application of hormone and azathioprine can cause obesity, osteoporosis, bone marrow suppression and increases the risk of concurrent tumor, etc. For patients with autoimmune hepatitis a more safe and reliable economic side effects of treatment is the purpose of the subject. Have reported between umbilical cord mesenchymal stem cells inhibit immunity, reduce inflammatory, inhibiting apoptosis, promoting proliferation function, its main is on T lymphocyte immune inhibitory effect, treatment for acute liver failure and end-stage liver disease. Between adipose mesenchymal stem cells also have multidirectional differentiation ability, large amount of specimen, based on simple, less pain and cell culture is stable. Based on the above background, this experiment to cultivate ectomesenchymal stem cell and lymphocyte culture together fat, restrain conditions and multiple factor correlation analysis of lymphocytes, and provide theoretical basis for clinical treatment of autoimmune hepatitis.objective:This study used ADMSCs and T cell co-culture analysis of different concentrations of ADMSCs inhibition of T cells And in the presence of IFN-r ADMSCs on expression of IDO and NF-KB activation condition and correlation of the two, In order to explore stem cells inhibit immune mechanism. If successful, can not only in the exploration of stem cells inhibit the immune mechanism, still can treatment of AIH to develop an efficient method to lay the foundation, to other types of liver disease andautoimmune diseases has important reference.Methods:5rats, female, weight between250g, KunMing medical university animal families and provide, under normal temperature raising, feeding the solid feed and drinking water. Repurchase of rats after30days in the new environment after being fat enough to start the experiment, the preoperative fasting12h, the water6h, weighing. Specim ens broken neck death aseptic conditions before open to the rat mesenteric adipose, collagenase digestion after separation of cells into the cultivation bottles in37℃, three days in liquid, regular observation of cell growth, and with the subculture, stay3to5generations grow well when take part for specificity between fat cells flow cytometry determined whether mesenchymal stem cell markers, and join the fluid cell directional differentiation into fat cells and nerve cells, the above identified as between adipose mesenchymal stem cells after cryopreserved for next step application. Rats50ml of blood lymphocyte separation medium separation T lymphocyte separation. Select the INF-r induced IDO and NK-kB between high expression of adipose mesenchymal stem cells, after24h of culture cell culture fluid collection, filtering, join the lymphocyte culture stimulation training, after48h, collecting cell for detection. Divided into four way.Group A blank control group Don’t join in blank control group lymphocyte high expression of adipose mesenchymal stem cell culture and SN50; Group B lymphocyte stimulation group to join in the high expression of adipose mesenchymal stem cell culture, not to join SN50; Group C block control group not join high expression in lymphocyte ectomesenchymal stem cell cultures to join SN50fat; Group D block lymphocyte stimulation group with high expression of adipose mesenchymal stem cell culture and SN50. After each collect lymphocytes and culture supernatant, PI dyeing processing lymphocytes, upflow cell meter flow detection, analysis, the change of the cell cycle. Cell culture supernatant on PCR and Western Blot detection of IDO and NF-kB molecules and protein expression level.Results:Between adipose mesenchymal stem cells grown in culture after15days is good, the high line of flow cytometry to detect specific indexes, and differentiate into fat cells and nerve cells, and between adipose mesenchymal stem cells and lymphocytes of the separation of the joint training, the results of group B compared with group A, C, D cells stay mostly in GO/G1phase, group B compared with group A, C, D IDO and NF-kB molecules and high level of protein expression (P≤0.05)Conclusion:Through adipose mesenchymal stem cells and lymphocytes between common culture determine the cell cycle and molecular and protein expression, can be obtained throu-gh the INF-r between adipose mesenchymal stem cells induced by gamma has inhib-ition to the lymphocyte, to autoimmune diseases in liver and gallbladder surgery prov-ides a theoretical basis for the treatment of autoimmune liver disease. |
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