[Specific expression in brain of exogenous gene by transpassing blood-brain barrier after intravenous injection].
OBJECTIVE
To study the expression of exogenous LacZ gene in brain via a delivery of OX26-pGFAP-IL.


METHODS
pCMV-Liposome, OX26-pCMV-IL, OX26-pGFAP-IL and blank liposome were injected into rats via femoral vein. At 24 h post-injection, the method of Q-PCR was adopted to calculate the relative quantities of LacZ gene mRNA in brain and peripheral organs. At 48 h post-injection, the protein expression of LacZ gene was detected by the activity of beta-galactosidase and the method of histochemical stain.


RESULTS
The result of Q-PCR showed that, at 24 h post-injection, the relative quantities of LacZ mRNA in OX26-pCMV-IL group (49.2 x 10(-6)) and OX26-pGFAP-IL group (44.9 x 10(-6)) were significantly higher than pCMV-liposome and blank liposome groups (P < 0.05). In peripheral organs, the relative quantity of LacZ mRNA in OX26-pCMV-IL group were significantly higher than that in OX26-pGFAP-IL group (P < 0.05). At 48 h post-injection, the activity of beta-galactosidase in OX26-pCMV-IL (0.67 pg/mg) and OX26-pGFAP-IL groups (0.92 pg/mg) were significantly higher than pCMV-liposome and blank liposome groups (P < 0.05). There was no significant difference between OX26-pCMV-IL group and OX26-pGFAP-IL group in terms of the expression of beta-galactosidase. The result of histochemical stain showed that OX26-pGFAP-IL achieved a specifically positive expression in brain and had a decreased expression in peripheral organs.


CONCLUSIONS
OX26-pGFAP-IL injected via femoral vein can cross the brain-blood barrier and achieve a specific expression in brain under the control of GFAP promoter. OX26-pGFAP-IL decreases the non-specific expression in peripheral organs and it may be used as an non-viral gene therapy for intra-cranial diseases.
Zhao
Hao
H
Department of Neurosurgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Peking Union Medical College, Beijing 100730, China.
Zhao
Ying-jie
YJ
Li
Gui-lin
GL
Wang
Ren-zhi
RZ
Kong
Yan-guo
YG
Feng
Ming
M
Wei
Jun-ji
JJ
Li
Shi-fang
SF
Ma
Wen-bin
WB
Yang
Yi
Y
Li
Yong-ning
YN
D000818:Animals; D001812:Blood-Brain Barrier; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007763:Lac Operon / Q000235:genetics*; D008081:Liposomes; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D008081:Liposomes; D001616:beta-Galactosidase