腺病毒介导的GDNF基因转移体外表达及生物学活性研究

Expression and Biological Activities of Adeno-Viral Mediated GDNF Gene Transfer in Vitro

目的 为利用重组腺病毒介导的胶质细胞源性神经营养因子 (GDNF)基因转移治疗帕金森病 (PD)提供依据。方法 采用免疫组化、RT-PCR及 ELISA定量分析观察人 GDNF腺病毒 (Ad-GDNF)在大鼠星形胶质细胞和 PC1 2细胞的表达 ,通过观察病毒直接感染及病毒感染的 PC1 2细胞上清对中脑原代培养细胞中的 TH阳性细胞 (DA能神经元 )生存能力和形态分化的影响来验证其生物学活性。结果 Ad-GDNF在星形胶质细胞、PC1 2细胞及大鼠中脑原代培养细胞均可有效表达 ,其表达产物对中脑 DA能神经元的生存和形态分化均有显著的促进作用。结论 腺病毒介导的 GDNF基因转移可在体外有效表达 ,且表达产物具有生物学活性 ,提示该手段在

Objective To provide basis for the application of adeno-viral mediated glia cell line-derived neurotrophic factor(GDNF) gene transfer in the treatment of Parkinson disease. Methods Immunohistochemical staining, RT-PCR and ELISA were performed to check the expressions of recombinant human GDNF adeno-virus(Ad-GDNF) in astrocytes, PC12 cells and primary mesencephalic cells for bioactivities of the expressive product of Ad-GDNF, direct Ad-GDNF infection or conditioned medium of Ad-GDNF infected PC12 cells on the survival and differentiation of tyrosine hydroxylase(TH)-positive cells (dopaminergic neurons) in the primary rat mesencephalic cultures. Results Ad-GDNF was expressed in astrocytes, PC12 cells and primary mesen-cephalic cells, The expressive products significantly promoted the survival and differentiation of dopaminergic neurons. Conclusions Adeno-viral mediated GDNF gene transfer was effectively expressed in vitro, and the expressive products was biologically active.The data from present study suggest that GDNF gene transfer for therapeutic approach can be highly helpful in the treatment of Parkinson disease.