【作者】 汪洋；

【导师】 周亚滨；

【作者基本信息】 黑龙江中医药大学 ， 中医内科学， 2012， 博士

【摘要】 目的：观察苏木乙酸乙酯提取物对同种异体心脏移植急性排斥反应模型大鼠供心组织NF-KB信号转导通路以及细胞毒性T淋巴细胞株CTLL-2凋亡的影响,探讨其免疫抑制作用的机制,为研发中药5类新药奠定工作基础。方法：将同种异体心脏移植急性排斥反应模型大鼠32只随机分4组,每组8只：环孢素A组、苏木加环孢素A半量组、苏木组、模型对照组,灌胃7天后采集供心,光镜观察供心组织病理变化；采用Western-blot测定供心组织NF-κBP65蛋白含量变化；Real Time-PCR测定供心组织IKB-α mRNA的表达。体外培养细胞毒性T淋巴细胞CTLL-2,细胞分为苏木乙酸乙酯提取物高、中、低剂量组及空白组,药物干预4.5h后,采用流式细胞术检测各组细胞凋亡情况。结果：(1)与模型对照组比较,环孢素A组、苏木加环孢素A半量组及苏木组能够明显减轻供心病理损伤,差异有统计学意义(P<0.05)；苏木加环孢素A半量组与环孢素A组减轻供心病理损伤作用相当,差异无统计学意义(P>0.05)。(2)与模型对照组比较,环孢素A组、苏木加环孢素A半量组及苏木组能够明显抑制供心组织NF-κBP65、IKB-α[的表达,差异有统计学意义(P<0.05)；苏木加环孢素A半量组与环孢素A组抑制供心组织NF-κB P65、IκB-α表达作用相当,差异无统计学意义(P>0.05)。(3)与空白组比较,苏木高、中、低剂量组均能促进细胞毒性T淋巴细胞凋亡,差异有统计学意义(P<0.05)；苏木高、中、低剂量组促进细胞毒性T淋巴细胞凋亡作用随着剂量的增大而增加,差异有统计学意义(P<0.05)。结论：苏木乙酸乙酯提取物能够抑制同种异体心脏移植急性排斥反应模型大鼠供心组织NF-κB信号转导通路的激活,并促进体外培养细胞毒性T淋巴细胞的凋亡,这可能是其免疫抑制作用机制之一。更多还原

【Abstract】 Objective:Obverse the influence of Sappanwood ethyl acetate extract to NF-kB signal transduction pathway of heart transplantation acute rejection allogeneic rats and apoptosis of CTLL-2T-cells.Discuss the mechanism of the immune inhibition function of Sappanwood ethyl acetate extractMethod:Device32heart transplantation acute rejection allogeneic rats to four groups randomly which contain Cyclosporine A group, mix group(half Sappanwood and half Cyclosporine A), Sappanwood group(Sappanwood ethyl acetate extract), control group(model control group). Every group has8rats. Irrigate the rats’stomach seven days, then collect donor heart tissue and observe pathological change of donor heart with light microscope. Assay protein content of NF-kB P65in donor heart tissue by Western-blot method. Assay content of IkB-α mRNA in Real Time-PCR method. Foster CTLL-2of T cell externally, divide cells to high dose group,middle dose group,low dose group and blank group,after intervene with medicine4.5hours,test apoptosis of every cell group with flow cytometry detection technique.Result:(1)Compare with control group, Cyclosporine A group, mix group and Sappanwood group alleviate pathological damage of donor heart obviously and there are statistical differences between them (P<0.05). Compare with Sappanwood group, Cyclosporine A group, mix group alleviate pathological damage of donor heart obviously and there are statistical differences between them (P<0.05). The alleviate pathological damage effect of donor heart of mix group and Cyclosporine A group are fairly.There are no statistical differences between them (P>0.05).(2)Compare with control group, Cyclosporine A group, mix group and Sappanwood group restrain the express of NF-kB P65and IkB-a of donor heart obviously and there are statistical differences between them (P<0.05). Compare with Sappanwood group, Cyclosporine A group, mix group restrain the express of NF-kB P65and IkB-a of donor heart obviously and there are statistical differences between them (P<0.05) The restrain effect of the express of NF-κB P65and IκB-α of donor heart of mix group and Cyclosporine A group are fairly.There are no statistical differences between them (P>0.05).(3)Compare with control group, high dose group, middle dose group and low dose group all can promote T-lymphocytes apoptosis and there are statistical differences between them (P<0.05).Compare each other group, with the increase of dose,the effect increased.There are statistical differences between them (P<0.05)Conclusion:Sappanwood ethyl acetate extract can restrain the activation of NF-κB signal transduction pathway and promote apoptosis of external foster T-cell.All these maybe one of the mechanisms of the immune inhibition function of Sappanwood ethyl acetate extract更多还原