Effects Of Alpha-Asarone On The Laminin Beta-1 In The Cell Model Of Intractable Epilepsy

OBJECTIVE To investigate the expression of laminin 1 gene and protein in the cell model of intractable epilepsy. Furthermore,to study the intervention effects of -asarone on the neuronal morphology and the laminin 1in this model and explore the antiepileptic mechanism of -asarone .METHODS: The neonate SD rats were decapitated quickly to obtain the hippocampal neurons, which were cultured in vitro in the serum-free neurobasal medium adding B27 supplement;At 7 day,the neurons were identified with neurofilament (NF) protein dyeing and calculated the purity;At 9 day,the neurons were randomly divided into control group(group A), model group(group B), model+7.5 g/ml -asarone group(group C), model+15 g/ml -asaronegroup (group D), model+30 g/ml -asarone group(group E), model+2 l ethanol group(group F),The phase-contrast microscope was used to observe the morphological changes of hippocampal neurons.The level of LDH(lactic acid dehydrogenase) in cell supernatant was measured at the time of 12h,24h and 48h by the automatic biochemistry analyzer. The expression of laminin 1 mRNA were examined by real-time fluorescence quantitative PCR, and the expression intensity of laminin 1 protein was examined by immunohistochemistry. RESULTS: (1)The neurons in the cell model of intractable epilepsy would occur and fusion. (2)A-asarone could reduce the activity level of LDH inthe cell culture medium. (3)At 12h in model group, the expression of laminin 1mRNA increased over time (P 0.05);At 24h in model group,the overexpression of laminin 1 mRNA was inhibited after -asarone was added,there was statistical significance of the difference between model group and model+ -asarone group P 0.05). (4)The expression of laminin 1 protein was mainly at the cytoplasm and matrix. AOD(average optical density): control group (0.0947±0.0259), model group (0.1351±0.0262), model+30 g/ml -asarone group 0.0979±0.0328 , there was statistical significance of the difference between control group and model group (P 0.01),also there was statistical significance of the difference between model group and model+30 g/ml -asarone group P 0.01)CONCLUSIONS:1 The gene and protein expression of laminin 1 increased in the cell model of intractable epilepsy,and there was morphologic change of neuronal network.(2) The cell model of intractable epilepsy existed neuronal damage , with time prolonging, the damage became more serious.(3) A-asarone could decrease the excessive expression of laminin 1 in the cell model of intractable epilepsy.