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目的:体外建立异体心肌组织抗原诱导大鼠淋巴细胞表面蛋白分子及编码基因表达时间窗模型,动态观察环孢素A对表达时间窗的影响。方法:实验分4组:对照组、抗原组、环孢组和抗原+环孢组。2、6、12、24、48h和72h观察脾白细胞其表面蛋白分子及编码基因的变化。结果:4组MHC-Ⅱ、CD4、CD8分子、ICAM-1无显著差异,CD4分子环孢霉素组2～24h有一个平台期。IL-2R分子抗原组12h达高峰值,环孢素A组6h有小峰值,抗原+环孢素A组表达较平稳。P59基因对照组逐步负调,而抗原组在2h内和12～24h有2个正调表达,2～12h负调至谷底,后再正调、负调表达,环孢素A组表达较稳定,抗原+环孢素组48h达到谷底,后迅速上升。CD4基因对照组2h、抗原组6h到谷底,后迅速上升,环孢素A组48h达到峰值,抗原+环孢素A组2h内有一个峰值,2组后逐步负调。CD8基因4组均负性表达,除抗原组无谷底外,对照组加药12h出现谷底,环孢素组加药6h出现谷底,抗原+环孢素A组加药12h出现谷底,3组后迅速上升。结论:异体心肌组织抗原可在2h内引发白细胞开始释放细胞表面蛋白分子IL-2R、P59基因表达,刺激6h可以诱发CD4基因开始转录表达,环孢素A 2h开始释放ICAM-I、IL-2R分子。这些结果为更加具体的描述移植免疫排斥反应的时间进程提供了详实的依据,并为更深入阐述环孢素A抑制移植排斥反应的机理提供了可靠依据。
OBJECTIVE: To establish a time window model of rat lymphocyte surface protein and gene expression induced by allogeneic myocardium antigens in vitro, and observe the effect of cyclosporine A on the time window of expression. Methods: The experiment was divided into 4 groups: control group, antigen group, cyclosporin group and antigen + cyclosporine group. 2, 6, 12, 24, 48h and 72h observed splenic white blood cell surface protein molecules and coding genes changes. Results: There was no significant difference in MHC-Ⅱ, CD4, CD8 and ICAM-1 between the four groups. The cyclosporine CD4 group had a plateau between 2 and 24 hours. IL-2R antigen group peaked at 12h, cyclosporin A group 6h had a small peak, antigen + cyclosporin A group was relatively stable. P59 gene control group gradually negative tone, and the antigen group within 2h and 12 ~ 24h two positive expression, 2 ~ 12h negative to the bottom, and then again, negative expression, cyclosporin A group was stable , Antigen + cyclosporine 48h reached the bottom, rose rapidly. CD4 gene control group 2h, antigen group 6h to the bottom, then rose rapidly, cyclosporine A group 48h peak, antigen + cyclosporin A group within 2h a peak, two groups gradually negative tone. CD8 gene 4 groups were negative expression, in addition to the antigen group without the trough, the control group at the bottom 12h after dosing, the bottom of the cyclosporine group at 6h, antigen + cyclosporin A group 12h after dosing bottom, after 3 groups Rising rapidly. CONCLUSION: Allogeneic cardiomyocytes antigens can initiate leukocyte release of IL-2R and P59 gene expression within 2h. The stimulation of 6h can induce the transcription of CD4 gene. The release of ICAM-I and IL-2R molecular. These results provide a more detailed basis for describing the time course of transplant rejection and provide a reliable basis for further elucidation of the mechanism of CsA inhibition of graft rejection.