The effect of different coating substrates on the expression of integrin α 2 in Sombati epileptic cell model

Abstract

Objective To investigate the effect of different coating substrates on the integrin α2 expression level of hippocampal neurons in Sombati epileptic cell model. Methods After coated with laminin and poly-L-lysine as substrates on the tissue culture plates respectively, hippocampal neurons were isolated from 24 h Kunming mouse and cultured in vitro. The primary cultured neurons were identified by immunohistochemical technique, and cultured with Mg(2+)-free extracellular fluid for 3 h to prepare Sombati epileptic cell model. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the integrin α2 mRNA expression of hippocampal neurons in each group on the eighth day. Results Before the model was prepared, neurons became full and large, processes connected closely, and neuronal network was formed at 5 d culture in vitro in laminin group, while the above appearances were seen at 7 d in poly-L- lysine group. Twenty-four hours after model preparation, neuronal migration phenomenon and "mesh-like" changes could be seen in all different groups. The ratio of the optical density of integrin α2 to β-actin could represent the relative expression level of integrin α2 mRNA. The ratio value in poly-L-lysine group, laminin group and poly-L-lysine + laminin group was 0.25 ± 0.03, 0.37 ± 0.05 and 0.48 ± 0.09, respectively. The expression level of integrin α2 mRNA in laminin group and poly-L-lysine + laminin group was all higher than that in poly-L-lysine group significantly (P = 0.005, 0.000; respectively). Conclusion In comparison of poly-L-lysine and laminin can strongly promote the formation of the neuronal network. Additional exogenous laminin can up-regulate integrin α 2 mRNA expression of hippocampal neurons in Sombati epileptic cell model.

DOI:10.3969/j.issn.1672-6731.2011.04.011