FasL基因转染对大鼠同种异体心脏移植的影响  

Effect of FasL transfection on the allograft of rat heart

作　　者：廖东山[1] 廖崇先[1] 陈良万[1] 陈昆[2] 余毅[2] 黄庆[2] 李增棋[2] 

机构地区：[1]福建医科大学附属协和医院心外科,福州350001 [2]福建医科大学附属第一医院心脏外科

出　　处：《中华实验外科杂志》2012年第8期1450-1452,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察转染FasL基因在心脏移植中是否可以诱导出一定程度的免疫耐受。方法取SD大白鼠为受体，Wistar大白鼠为供体，各68只，分成3组，I组：20只，心脏直接进行移植：Ⅱ组：20只，单用多聚乙烯亚胺（PEt）转染心脏后移植；Ⅲ组：28只，供心移植前用PEi介导克隆大鼠FasL（rFasL）基因转染，质粒DNA（肛g）：PEI（Ixl）=20：40。分别检测移植心脏存活时间、心肌组织的苏木素一伊红（HE）染色、FasL的表达、原位末端转移酶标记（TUNEL）法检测心肌组织的原位细胞凋亡、Westernblot法检测FasL蛋白水平。结果（1）Ⅲ组移植心脏的存活时间为（17．8±0．6）d，较另2组明显延长（P〈0．01）。（2）I组和Ⅱ组心肌组织的FasL水平渐升高，但前3d均未超过40％，Ⅲ组FasL水平在移植后3d即达较高水平（63．65±3．31）邯％（P〈0．01）。（3）随着移植时间的延长，Ⅲ组心肌组织细胞凋亡指数由（13．40±3．46）细胞数／mm2最终升至（48．90±6．91）细胞／mm2（P〈0．05），凋亡细胞大多为浸润的炎性细胞，但I组和Ⅱ组有大量的心肌细胞凋亡。（4）Ⅲ组FasLmRNA的转录强度和蛋白表达均比Ⅰ、Ⅱ组明显升高。结论适当浓度的FasL基因转染移植心脏，可以诱导出一定程度的免疫耐受。Objective To observe the effects of FasL trausfection on immunological tolerance in a certain extent in heart transplantation. Methods We chose the SD rats and the Wistar rats as recipients （n = 68 ） and donors （n = 68 ） respectively. All the rats were divided into 3 groups： 20 in group I , the donor hearts were transplanted directly; 20 in group II , the donor hearts were transfeeted only by polyeth- yleuimine （PEI） without rFasL transfection ; 28 in group III, DNA plasmid （pg） ： PEI （ pl ） = 20 ： 40. The survival time of the graft, the hematoxylin and eosin （HE） stain of the heart tissue, the expression of rFasL mRNA in the heart tissue, in situ apoptosis of the heart tissue with TdT-mediated dUTP nick end la- beling （TUNEL） , the rFasL protein with Western blotting were assessed by using statistic software SPSS 18.0. The difference was significant if the value of P was lower than 0. 05. Results （ 1 ） The survival time of the grafts in group m was the longest （ 17.8 ±0. 6） days. There was significant difference in the survival time of the grafts between group III and groups I , II （both P 〈0. 01 ） ; （2） The expression of rFasL in the heart reached the peak at the 3rd day post-transplantation in group III （63.65 ± 3.31 ） ＂q% , but never exceeded 40% in groups I and II at the first 3 days post-transplantation （P 〈 0. 01 ） ; （3） The apoptotic index of heart tissue in group III was increased from （ 13.40 ±3.46） cells/ram2 on the day 1 to （ 13.40 ±3.46） cells/ram2 on the day 7, and higher than thoat in groups I and II （P 〈0. 05）. There were more apoptotic myocardial cells in groups I and II , but inflammatory cells in group III; （4） The expression of rFasL protein in group III was stronger than that in groups I and II. Conclusion FasL transfection into the heart graft with appropriate concentration can induce immunological tolerance in a certain extent.

关 键 词：心脏移植 多聚乙烯亚胺 FASL基因 基因转染 

分 类 号：R654.2[医药卫生—外科学]