反义脱氧寡核苷酸静脉输注抑制小鼠MD-1表达延长皮肤移植物存活时间的研究  

Inhibition of MD-1 expression by antisense oligodeoxynucleotides and its impact on skin allograft survival of mouse

作　　者：张鑫[1] 陈必成[1] 昌盛[1] 周鸿敏[1] 杜敦峰[1] 袁劲[1] 陈忠华[1] 

机构地区：[1]华中科技大学同济医学院同济医院器官移植研究所国家教育部器官移植重点实验室国家卫生部器官移植重点实验室,武汉430030

出　　处：《中华微生物学和免疫学杂志》2005年第9期733-736,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家973基金资助项目(2003CB515505)

摘　　要：目的静脉输注反义脱氧寡核苷酸(简称反义核酸,AS-ODN)抑制小鼠MD-1表达,并初步探讨MD-1用于排斥反应诊断的前景。方法实验组(组1和组2)和阳性对照组(组3)行C57BL/6-BALB/c同种皮肤移植,阴性对照组(组4)行BALB/c-BALB/c同基因皮肤移植后,分别静脉输注小鼠MD-1AS-ODN(组1)、随机序列脱氧寡核苷酸(组2)或生理盐水(组3和组4)。于术后第11天留样检测脾细胞MD-1表达水平(FACS)、脾细胞增殖反应强度(AlamarBlue还原率)、血清IL-2和IL-10水平(ELISA)以及皮肤移植物组织学改变(组织病理学检查),并记录移植物存活时间。结果静脉输注小鼠MD-1AS-ODN能明显减少BALB/c小鼠脾细胞中MD-1阳性细胞百分数,降低脾细胞对特异性和非特异性刺激的增殖反应强度,下调血清IL-2水平和上调IL-10水平,减少移植物局部淋巴细胞浸润,并显著延长同种皮肤移植物存活时间。结论静脉输注MD-1AS-ODN特异性抑制小鼠MD-1表达可显著延长皮肤移植物的存活时间。MD-1表达水平对排斥反应的发生具有一定的指示作用。Objective To study the effects of specific antisense oligodeoxynucleotides （AS-ODN）on MD-1 expression and skin allograft survival. Methods Group 1, AS-ODNs and group 2, random ODN（R-ODN）, were administrated by 100 μg/48 h ×8 doses i. v, whereas the animals in group 3（positive control） and group 4 （negative control） were given normal saline by 0.5 ml/48 h × 8 doses i.v. C57BL/6 to BALB/c skin allograft model was employed in groups 1-3, and BALB/c to BALB/c skin graft model was employed in group 4, followed by the defined treatments from day 0. Recipients were sacrificed at day 11 after transplantation. The expression of MD-1 on splenocytes was determined by FACS. AlamarBlue was used to evaluate the proliferation of recipient splenocytes. The serum levels of IL-2 and IL-10 was detected by ELISA. Results The mean survival time（MST） of skin allografts in group 1 was significantly prolonged, but R-ODN treatment had no effect on it as compared with group 3（ P 〈 0.05）. AS-ODN inhibited MD-1 expression and lymphocyte proliferation, down-regulated serum level of IL- 2 and of IL-10, but R-ODN treatment showed no significant influence upon these（ P 〈 0.05, vs controls）. The grafts from treatment groups showed clear tissue structure with very few lymphocytes, while group 2 showed severely histological lesions of rejection. Conclusion AS-ODN treatment efficiently inhibited MD- 1 expression and significantly prolonged the survival time of skin allografts. The expression level of MD-1 may function as an indicator for rejection.

关 键 词：MD-1 反义脱氧寡核苷酸 细胞因子 增殖反应 存活时间 反义脱氧寡核苷酸 移植物存活时间 皮肤移植物 小鼠脾细胞 MD-1 静脉输注 显著延长 非特异性刺激 脾细胞增殖 

分 类 号：R965[医药卫生—药理学]