Acta Pharmacologica Sinica 2005 January; 26 (1): 33-38; doi: 10.1111/j.1745-7254.2005.00002.x

Acta Pharmacologica Sinica 2005 January; 26 (1): 33-38; doi: 10.1111/j.1745-7254.2005.00002.x

Original Article

Antiapoptotic effect both in vivo and in vitro of A20 gene when transfected into rat hippocampal neurons¹

Hong-sheng MIAO^2,3, Lu-yang YU⁴, Guo-zhen HUI², Li-he GUO^2,5

² Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China;³ Department of Neurosurgery, First Hospital of Soochow University, Suzhou 215007, China

Aim: To evaluate the antiapoptotic effect of the A20 gene in primary hippocampal neurons both in vivo and in vitro.

Methods: Primary hippocampal neurons in embryonic day 18 (E18) rats were transfected with the A20 gene by using the new Nucleofector electroporation transfection method. We then examined, whether A20 -neurons possessed anti-apoptotic abilities after TNF-α stimulation in vitro. A20-neurons and pcDNA3 -neurons were transplanted into the penumbra of the brains of rats that had been subjected to 90-min of ischemia induced by left middle cerebral artery occlusion (MCAO).

Results: A20-neurons resisted TNF-α induced apoptosis in vitro. The apoptosis rate of neurons overexpressing A20 (28.46%±3.87%) was lower than that in neurons transfected with pcDNA3 (53.06%±5.36%). More A20-neurons survived in the penumbra both 3-d and 7-d after transplantation than did sham pcDNA3 neurons.

Conclusion: The novel function of A20 may make it a potential targets for the gene therapy for neurological diseases.

Keywords: A20 protein; tumor necrosis factor; primary hippocampal neurons; middle cerebral artery infarction; apoptosis

¹Project supported by Major State Basic Research Development Program of China (No G1999053905).

^{2 ,4}These two authors contribute equally to this work.
⁵Correspondence to Prof Li-he GUO.
Phn/Fax 86-21-5492-1391.
E-mail lhguo@sibs.ac.cn
Received 2004-05-26 Accepted 2004-09-17

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