| 【 Objective 】 By the injection of imDC induced by sinomenine to the recipient rats,observed the Th1 / Th2 cytokines in venous blood of the five groups receptors, did reseach of the effects on Th1 / Th2 cytokines through the imDC induced by sinomenine,to probe into the mechanism which imDC induced by sinomenine can lead transplantation immune tolerance,to provide theoretical basis and experimental basis for applying sinomenine to induce immune tolerance of renal transplantation.【 Methods 】 Took wistar rats bone marrow-derived precursor cells,then divided them into group A and group B,group A used the conventional inducement method,which used the medium which contain the FBS of 10% and GM-CSF of 10ng/ml to induce the bone marrow cells of Wistar rat,group B used the method which induced by sinomenine that add the sinomenine of 100 u M,the rest of them were the same as the group A.Cultivated a large number of immature dentritic cells,to observe the growth of the cells and flow cytometry cell phenotype.Chose the inbred strain SD rats as the renal transplant recipients;Chose the inbred strain Wistar rats as kidney donor,Wistar'SD.Used the random allocation to establish the animal models,divided the SD rats into three groups,each group had twelve rats.Group A:comparison group,trickled the normal saline into the rat caudal vein seven days befor the renal transplantation;Group B : imDC105 group,trickled the im DC of 105 which had not been induced by sinomenine into the rat caudal vein seven days befor the renal transplantation; Group C: imDC106 group,trickled the imDC of 106 which had not been induced by sinomenine into the rat caudal vein seven days befor the renal transplantation;Group D:imDC105-SN group,trickled the imDC of 105 which had been induced by sinomenine into the rat caudal vein seven days befor the renal transplantation;Group E :imDC106-SN group,trickled the imDC of 10 6 which had been induced by sinomenine into the rat caudal vein seven days befor the renal transplantation;.Used the vessel sutures of the microsurgery technique to build the bilaterally renal transplantation model of rats,observed the survival time of each group after the operation,extracted six recipient from each group to draw blood from their caudal vein seven days after the renal transplantation,used the ELISA method to determine the lever of the IL-2、IL-4、IL-10 and INF-γ.【Results】1. The conventional induction group and sinomenine induction group had the following characteristics:① Dendritic cell morphology under a microscope had characteristic of a lack of maturity. characteristic of; by flow cytometry dendritic cell phenotype C② D80 and RT1 B were in low expression detected by flow cytometry;Under the effects of various factors could be induced to produce a number of rat bone marrow-derived immature dendritic cells which could meet the requirement of the subsequent experiments.2. By the using of the vessel sutures of the microsurgery technique to build the singel renal transplantation model of rats,the harvesting time was 28.3±5.6min,the arterial anastomosis time was 12.5±5.7min,the venous anastomosis time was 17.3±3.4min,the success rates could be 89.5 %, which could meet the need of the following experiment.3.In the detecting of the Th1 / Th2 cytokines in venous blood of the five groups receptors, the lever of the IL-4 and IL-10 which stood for the Th2 cytokines should be :among the same concentration,imDC-SN group was higher than imDC group and comparison group,the difference had statistic significance(P<0.05); imDC106-SN group was higher than im DC105-SN group, the difference had statistic significance(P<0.05);the lever of the IL-2 and INF-γ which stood for the Th1 cytokines should be :among the same concentration,imDC-SN group was lower than imDC group and comparison group,the difference had statistic significance(P<0.05); imDC106-SN group was lower than imDC105-SN group,the difference had statistic significance(P<0.05);In imDC106-SN group and imDC105-SN group,the lever of the IL-4 and IL-10 which stood for the Th2 cytokines is higher than the other groups,the lever of the IL-2 and INF-γ which stood for the Th1 cytokines is lower than the other groups..【Conclusions】1.Using the method of microsurgery to anastomosis the blood could make the model of singel kidney transplantation in rats,which met the requrement of the following experiment.2.The experiment proved that:specificity imDC induced by sinomenine could rise the lever of Th2 cytokines and drove down the lever of Th1 cytokines,induced the migration to Th2 immune response,the effect of imDC-SN group was stronger than the other groups and the effect became stronger when the concentration of sinomenine rose,which proved im DC induced by sinomenine could mediate immune tolerance to the recipient. |
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