| Objective: To further explore the immunosuppressive mechanism of SIN(Alkaloid sinomenine) in organ transplantation by studying its effects on expression levels of CD4+CD25+Treg( CD4+CD25+Regulatory T Cells, Treg) and Foxp3 in spleens of rats undergoing kidney transplant, so as to provide experimental basis for immunosuppressive effects of SIN on organ transplantation.Materials and Methods: Inbred strain SD rats were selected as kidney transplant recipient; inbred strain Wistar rats were selected as donor, and all of them were male rats with a body weight of about250-300 g. Randomized block design was used to set up animal models;SD rats(recipient) were divided into 5 groups, with 12 rats in each group.A The control group: rats were injected with normal saline via their tail veins 7 days before kidney transplant; B the im DC105 group: rats were injected with 1*105 im DC suspension that hadn’t been treated with SIN via their tail veins 7 days before kidney transplant; C the im DC106 group:rats were injected with 1*106 im DC suspension that hadn’t been treated with SIN via their tail veins 7 days before kidney transplant(10^6/rat); D the im DC105-SN group: rats were injected with 1*105 im DC suspension that had been treated with SIN via their tail veins 7 days before transplant;E the im DC106-SN group: rats were injected with 1*106 im DC suspension that had been treated with SIN via their tail veins 7 days before transplant. The acute kidney-transplant rejection model was established by orthotopic left kidney transplantation, from Wistar rats to SD rats. After transplant, these rats were killed after being raised in clean grade-environment for 7 days and their spleens were taken to make suspension; percentages of CD4+CD25+-Treg and CD4+CD25+Foxp3+-Treg in CD4+T cells were examined by flow cytometry; after getting the data, the SPSS18.0 software was used to process and analyze the data.Results: 1. Flow cytometer was used for count of CD4+CD25+-Treg cells in spleen: A the control group: 4.5±1.0; B the im DC105 group:3.9±0.6; C the im DC106 group: 7.4±1.1; D the im DC105-SN group:10.8±1.7; E the im DC106-SN group:14.9±1.7. There was significant difference that the E group significantly higher than the A group and the C group(P<0.05), the D group than the A group and the B group is too.And among these groups, apart from that there was no significant difference between either two of the A group, the B group and the C group(P>0.05),there was significant difference between the E group and the D group(P<0.05). Proliferation rates of Treg cells from high to low were as follows: the E group, the D group, the C group, the A group and the B group. 2. Flow cytometer was used for count of Foxp3 :(1) the control group: 3.6±0.7;(2) the im DC105 group: 2.8±0.8;(3) the im DC106group: 6.3±0.3;(4) the im DC105-SN group: 9.1±1.3;(5) the im DC106-SN group: 11.6±1.8. There was significant difference that the E group significantly higher than the A group and the C group(P<0.05), the D group than the A group and the B group is too.And among these groups,apart from that there was no significant difference between either two of the A group, the B group and the C group( P > 0.05),there was significant difference between the E group and the D group( P <0.05).The B group had the lowest count of Foxp3, while the E group had the highest count of Foxp3.Conclusions:1. Proliferation of CD4+CD25+Treg cells in spleen of rats undergoing kidney transplant were promoted by SIN-treated im DC in a concentration dependent way;2. Proliferation of Foxp3 in spleen of rats undergoing kidney transplant were promoted by SIN-treated im DC in a concentration dependent way;3. By maintaining im DCs from the donor in an immature state, SIN could promote the proliferation of Treg cells in rats undergoing kidney transplant and up-regulate the expression level of Foxp3+Treg, thus to realize its immunosuppressive effect on transplant. |
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