Bcl-2 gene therapy for apoptosis following traumatic brain injury.
OBJECTIVE
To investigate the therapeutic effect of Bcl-2 fusion protein on apoptosis in brain following traumatic brain injury.


METHODS
Bcl-2 gene was cloned by RT-PCR. Bcl-2 and EGFP genes were linked together and inserted into pAdeno-X vector. This recombinant vector was packaged into infectious adenovirus in HEK293 cells. Ninety Wistar rats were assigned randomly into experimental group (n=45) and control group (n=45). All rats were subjected to traumatic brain injury. Then recombinant adenovirus (for experimental group) or saline (for control group) was injected into the traumatic brain. The expression of Bcl-2 fusion protein was investigated by Western blotting, immunohistochemistry and fluorescence microscopy. Apoptosis in the injured brain was studied by TUNEL. Animals' behavior capacity was evaluated by tiltboard test.


RESULTS
In the experimental group, many fluorescent cells were found around the traumatic locus, which were also proven to be Bcl-2 positive by immunohistochemistry. On the contrary, few Bcl-2 positive cells and no fluorescent cell were detected in the control group. Bcl-2 expression of experimental group was much higher than that of control group, which was illustrated by Western blotting. The apoptosis index of experimental group was 0.027+/-0.005, and that of control group was 0.141+/-0.025 (P < 0.01). Two weeks after injury, animals of the experimental group behaved better than those of the control group.


CONCLUSIONS
A recombinant adenovirus vector expressing Bcl-2 fusion protein has been constructed. Bcl-2 fusion protein can suppress apoptosis and promote cell survival. Moreover, the behavior recovery of the injured animal is promoted. Bcl-2 fusion protein provides a way to track the target cells in vivo.
Yang
Xiao-feng
XF
Department of Neurosurgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China.
Zheng
Xue-sheng
XS
Liu
Wei-guo
WG
Feng
Jun-feng
JF
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D001930:Brain Injuries / Q000628:therapy*; D003001:Cloning, Molecular; D019254:Genes, bcl-2; D015316:Genetic Therapy; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar
D019253:Proto-Oncogene Proteins c-bcl-2