摘要
目的观察人α1，2-岩藻糖苷转移酶（HT）和衰变加速因子（DAF）基因转移对抑制血管内皮细胞激活从而克服异种移植急性血管排斥反应的能力。方法通过显微注射建立转基因小鼠动物模型，Southern印迹杂交和流式细胞计数筛选出人HT和／或DAF基因整合与表达阳性的子代转基因小鼠。研究表达不同目的基因的转基因小鼠血管内皮细胞与15％人血清孵育后溶破细胞的百分数，免疫细胞化学染色检测细胞核因子（NF）-KB的激活，流式细胞计数检测细胞表面血管细胞黏附分子（VCAM）-1的表达。结果子代小鼠血管内皮细胞人HT／DAF共基因表达7只，人HT与DAF单基因表达分别为6只和8只。与15％人血清孵育后，人HT／DAF共表达组细胞的溶破细胞百分数（4±2）％低于人HT表达组细胞（25±10）％、人DAF表达组细胞（31±11）％及正常组细胞（76±24）％（P均〈0．05）。转双基因抑制细胞NF—KB激活的能力强于转人HT或DAF单基因，且转双基因细胞表面VCAM-1的表达较转单基因细胞显著降低，差异有统计学意义（P〈0．05）。结论人HT／DAF基因联合转移具有部分抑制血管内皮细胞激活从而克服异种移植急性血管排斥反应的能力。
Objective To investigate the role of human α （1,2）-fucosyhransferase （HT） and decay accelerating factor （DAF） in overcoming xenograft acute vascular rejection by inhibiting vascular endothelial cells （Ecs） activation. Methods Transgenic mice were produced by microinjection of transgene constructs for human HT and DAF. Integration and expression of human HT and DAF transgenes were tested by Southern blotting and flow cytometry （FCM）. Murine vascular ECs were exposed to 15% human serum,then the percentage of lyzed cells were determined. Immunohistochemistry was used to detect the activation of nuclear factor-kappa B （NF-kB）. FCM was used to analyze the expression of VCAM-1 in ECs. Results Among offspring generation mice, the expression of human HT/DAF, HT or DAF was 7,6 and 8 respectively. After incubated with human serum, the percentage [ （4 ± 2 ） % ] of lyzed cells of ECs co-expressing human HT/DAF was lower than that of ECs expressing human HT [ （25± 10） % ] or DAF [ （ 31 ± 11 ） % ] （ all P 〈 0.05 ）. Co-expression of human HT/DAF gene in ECs could inhibit the activation of NF-KB and reduce the VCAM-1 expression more efficiently than those expressing either human HT or DAF gene. Conclusion All the data suggests that co-expression of human HT/DAF could inhibit vascular ECs activation, and might be a useful method to overcome xenograft acute vascular rejection.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第6期780-782,共3页
Chinese Journal of Experimental Surgery
关键词
异种移植
急性排斥反应
衰变加速因子
基因表达
Xenotransplantation
Acute rejection
Decay accelerating factor
Gene expression
