Construction of polybutylcyanoacrylate nanoparticles loaded brain derived neurotrophic factor gene and their expressions in rat brain tissues
10.3760/cma.j.issn.1671-8925.2015.10.001
- VernacularTitle:聚氰基丙烯酸正丁酯纳米粒载BDNF基因的制备及在活体大鼠脑内表达的实验研究
- Author:
Xue LAI
1
;
Wu ZHONG
;
Yingchun HU
;
Hao LI
;
Yu XIONG
;
Ligang CHEN
Author Information
1. 四川医科大学附属第一医院急诊医学部
- Keywords:
Polybutylcyanoacrylate nanoparticle;
Brain derived neurotrophic factor;
Craniocerebral injury
- From:
Chinese Journal of Neuromedicine
2015;14(10):973-978
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare the polybutylcyanoacrylate nanoparticles (PBCA-NPs) loaded brain derived neurotrophic factor (BDNF) gene as the gene delivery system and explore their expressions in rat brain tissues so as to observe the influence of PBCA-NPs in BDNF expression.Methods PBCA-NPs were prepared by emulsion polymerization method.Surface of PBCA-NPs was surveyed by transmission electron micrograph (TEM) and zeta potentials of PBCA-NPs were determined with laser grain analyzer.The PBCA-NPs surface was modified by cationic surfactant cetyltrimethylammonium bromide (CTAB).The eukaryotic expression vectors PPEGFP-BDNF were constructed;after verification by double enzyme digestion and sequencing, pPEGFP-BDNF was packaged by PBCA-NPs.Forty-eight male SD rats were randomly divided into blank-control group, PBCA group, pPEGFP-BDNF group and PBCA-PEGFP-BDNF group (n=12), and Feeney's method was used to induce craniocerebral injury models in these rats, and then, one mL normal saline, PBCA-NPs, PEGFP-BDNF plasmids and PBCA-PEGFP-BDNF plasmids were given to the above groups.Seven d after that, peripheral brain tissues of right injury brain tissues were chosen;expressions of BDNF gene were detected by pathological examination, real time-PCR and Western blotting.Results Nps with even size and smooth surface were successfully obtained, holding the high zeta electric potential ([62.23±2.15] ％).The new constructed vectors were confirmed by restricted enzyme and sequencing.Real time-PCR indicated significant difference of BDNF mRNA expressions among the four groups (F=112.668, P=0.000);as compared with that in the other three groups, the BDNF mRNA expression in the PBCA-PEGFP-BDNF group was significantly higher (P＜0.05).Western blotting indicated significant difference of BDNF protein expressions among the four groups (F=66.629, P=0.000);as compared with that in the blank group and PBCA group, the BDNF protein expression in the PEGFP-BDNF group was significantly higher (P＜0.05), and that in the PBCA-PEGFP-BDNF group was significantly higher than the other three groups (P＜0.05).Conclusion PBCA-NPs could be a good vector and provide a new way for gene therapy of craniocerebral injury.
