环孢素对心脏移植大鼠外周血淋巴细胞多基因mRNA表达的影响

 目的 对大鼠心脏移植前后不同时间点外周血淋巴细胞mRNA差异表达的研究,探讨环孢素对免疫排斥反应过程中基因表达的影响.方法 心脏移植前后以15 mg·kg^-1·d^-1环孢素灌胃的大鼠,分别在移植前、移植后24,72 h,7,10,12d取血,提取总RNA,用mRNA差异显示法找出差异表达条带,克隆测序,用灰度扫描法计算差异表达基因的比密度。结果 对照组移植心脏存活率于移植后的10d内降至为0%,而此阶段环孢素组的移植心脏存活率始终保持在50％.移植后第10天,对照组与凋亡信号传导和线粒体蛋白构象重构与蛋白转运相关基因Ribosomal—12S和S25,MHC—Ⅲ(Hst-70)的表达水平显著高于移植前,与移植心脏存活率最低点同步.其2,3-bisphosphoglycerate和gig18基因表达水平的变化无统计学意义。环孢素组与细胞凋亡有关的gig18和Thbsl,以及T细胞受体(TCR)等基因的表达水平于移植后10 d内持续低于移植前,2,3-bisphosphoglycerate基因的表达水平于移植后24 h降低,只有钠通道β-1基因的表达水平于移植后10 d内高于移植前。结论 环孢素可负调心脏移植后大鼠外周血淋巴细胞凋亡相关基因gig18和Thbsl,正调钠通道基因的转录。

Effects of ciclosporin A on mRNA expression of peripheral lymphocytes in rats undergoing heart transplantation

OBJECTIVE To investigate the effect of ciclosporin A (CsA) on the gene expression of peripheral lymphocytes in rats undergoing heart transplantation. METHODS The rats were divided into control group and CsA group. The rats in CsA group were orally administered CsA 15 mg.kg-1.d-1 from 4 d before transplantation to 12 d post-operation. Blood were collected at pre-transplantation and at 24, 72 h, 7, 10 and 12 d post-operation. Lymphocytes were isolated and (RNA was prepared. Differential display and sequencing technio,ues were used to assess multiple genes expression level. RESULTS The survival rate of transplanted heart of the control group was reduced to 0% at 10 d post-transplantation. At the same time, the expression of rihosomal-12S/S25 and MHC-III , which related to apoptosis signal transmission and protein configurational reconstruction and transportation in mitochondrion, reached the maximum value. However, no statistical significance was found for the change of 2,3-bisphosphoglycerate and gig18 genes expression. In CsA group, the survival rate of transplanted heart maintained at 50% from 3 to 10 d post-operation. The expression of gig18, Thbs1 and TCR were down-regulated during this period, while that of sodium channel beta-1 was up-regulated. CONCLUSION CsA down-regulated the express of apoptosis-relating genes of gig18 and Thbsl, and up-regulated the express of sodium channel gene in peripheral lymphocytes in rats undertaken xenogenous heart transplantation.