Rapamycin和cyclosporin A对同种排斥过程中TLR5及Foxp3表达的影响

Effect of Rapamycin and Cyclosporin A on the expression of TLR5 and Foxp3 in allorejection

目的:研究雷帕霉素(Rapamycin,RAPA)和环孢素A(cyclosporin A,CsA)体内处理对同种移植小鼠急性排斥过程中TLR5及Foxp3表达的影响。方法:建立同种皮肤移植模型,术后给予RAPA或CsA,1次/d,连续14d,同时设生理盐水对照组。每日观察移植物存活状况。术后选取1、7、10、14、21d共5个时间点,提取受体小鼠脾细胞,RT-PCR检测TLR5及Foxp3mRNA表达,探讨不同处理组基因表达与移植物生存的相关性。体外实验利用鞭毛蛋白(flagellin)联合RA-PA和CsA处理正常小鼠T细胞6h,RT-PCR检测TLR5表达。结果:RAPA和CsA可明显延长小鼠同种移植皮片存活期。RAPA可促进脾细胞TLR5mRNA表达升高,停药后的第21天仍明显高于CsA组和对照组(P<0.05);CsA组在第7、10天有显著升高,第21天降低(P<0.05);3组中最高表达的时间点为移植后第10天,此时正值排斥高峰期。RAPA可引起Foxp3mRNA的表达升高(P<0.05),停药后仍维持较高水平;CsA组仅在移植后第7、10天短暂升高,第14天低于对照组(P<0.05)。移植后1、7、10、21d3组的TLR5与Foxp3mRNA变化趋势正相关。体外实验中,RAPA或CsA与flagellin联合使用,可促进TLR5的表达,以前者的作用更为明显。结论:RAPA和CsA在同种移植急性排斥高峰期可引起TLR5和Foxp3表达的协同升高,且RAPA的作用更加明显和持久,鞭毛蛋白体外可以增强这种作用效果。

AIM： To study the effect of Rapamycin （RAPA） and Cyclosporin A （CsA） on the expression of TLR5 and Foxp3 in allotransplantation model in vivo. METHODS： The murine model of skin allotransplantation was established, and divided into three groups, injected with CsA 10 mg/（kg·d）, RAPA 1.5 mg/（kg · d）） and normal saline respectively for 14 consecutive days. The grafts survival was observed daily. The expression of TLR5 and Foxp3 mRNA was detected by RT-PCR. RESULTS： The survival time was obviously longer in RAPA and CsA treated groups than control group. The expression of TLR5 mRNA in allotransplantation treated with RAPA was higher than that in control group （ P 〈 0.05） and remained at a higher level than the other two groups until day 21. TLR5 showed the highest expression on day 10 among three groups, which was the top point of allorejection. Compared with CsA group, RAPA treatment in vivo caused the higher expression of Foxp3 mRNA （ P 〈 0.05）, which remained at a high level after treatment stopped. However, in CsA group, Foxp3 mRNA expression increased on day 7 and 10, and then decreased significantly on day 14. TLR5 and Foxp3 expression was positively correlated among three groups. RAPA and CsA promoted the expression of TLR5 in T cells when treated with flagellin for 6 hours in vitro and RAPA has stronger effect. CONCLUSION： RAPA and CsA can promote the expression of TLR5 and Foxp3 in ailotransplantation model in vivo and flagellin enhanced this effect in vitro.