创伤性脑损伤后细胞凋亡及Bel—2基因治疗  

Cell apoptosis and Bcl-2 gene therapy following traumatic brain injury

作　　者：冯志仙[1] 杨小锋[1] 郑学胜[1] 刘伟国[1] 潘德生[1] 罗鸣[1] 沈罡[1] 沈方[1] 

机构地区：[1]浙江大学医学院附属第一医院神经外科,杭州310009

出　　处：《中华创伤杂志》2007年第8期561-564,共4页Chinese Journal of Trauma

摘　　要：目的 研究创伤性脑损伤（TBI）后的细胞凋亡情况，并探讨Bcl-2基因治疗对凋亡的作用。方法 60只SD大鼠随机分成TBI组（15只）、假处理组（15只）、基因治疗组（15只）和空载体对照组（15只）。除了假处理组外，所有大鼠均用Feeney自由落体致伤装置制成TBI模型。通过RT—PCR法克隆Bcl-2基因cDNA，并利用重组真核表达载体转染受损的脑细胞。用免疫荧光法检测Bcl-2基因的表达。原位末端标记法和电镜检测细胞凋亡情况。结果 与假处理组相比，TBI组的伤灶周围可以发现更多凋亡细胞，并且两者之间的凋亡指数差异有统计学意义（P〈0．01）。通过脂质转染，Bcl-2基因成功在体内表达。基因治疗组的Bcl-2（＋）细胞的标记指数为（8．3±0．7）％，而在空载体对照组中仅为（0．9±0．2）％（P〈0．01）。结论 细胞凋亡存在于损伤的脑组织中，而Bcl-2基因治疗能抑制这种凋亡的发生，后者为TBI后早期干预促进脑细胞存活提供了一种可行的方法。Objective To study the cell apoptosis following traumatic brain injury （TBI） and explore the therapeutic effect of Bcl-2 gene therapy on it. Methods A total of 60 SD rats were assigned randomly to TBI group （ n = 15 ）, sham control group （ n = 15 ） ,gene therapy group （ n = 15 ） and Empty- Vec control group （ n = 15 ）. Except for sham control group, rats were subjected to severe TBI by Feeney＇ s method. The cDNA of Bcl-2 gene was cloned by RT-PCR. A recombinant eukaryotic expression vector was constructed to transfect the injured brain cells and Bcl-2 expression in vivo was detected by immunofluorescence. The cell apoptosis of each group was detected by TUNEL and electron microscopy. Results Compared with sham control group, more apoptotic cells were found around the traumatic focus in TBI group, with a significant statistical difference in the apoptosis index between two groups （P 〈 0.01 ）. Bcl-2 was expressed successfully in vivo after lipofection. The Bcl-2 （ ＋ ） cell labeling index was （8.3 ＋ 0.7） % in gene therapy group and only （0.9 ＋ 0. 2 ） % in Empty-Vec group （ P 〈 0. 01 ）, which suggested that enhanced expression of Bcl-2 resulted in a decline in apoptosis. Conclusions It is found in the present study that apoptosis occurs following TBI. Bcl-2 expression in vivo can inhibit apoptosis, which provides a practicable way for early intervention to promote cell survival.

关 键 词：脑损伤 创伤性 基因 BEL-2 基因治疗 凋亡 

分 类 号：R686[医药卫生—骨科学]