α-细辛醚对难治性癫痫细胞模型及Laminin β_1的影响  

Effects of α-Asarone on the Cell Model of Intractable Epilepsy and Gene of Laminin β_1

作　　者：马美刚[1] 吴原[1] 吴月娟[1] 苏婕[1] 刘秀颖[1] 唐玉兰[1] 余璐[1] 

机构地区：[1]广西医科大学第一附属医院,广西南宁530021

出　　处：《时珍国医国药》2011年第5期1059-1061,1064,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30960111);广西自然科学基金(No.2010GXNSFA013168);广西医疗卫生重点课题(桂卫重200916);广西2010年研究生教育创新计划资助项目(No.2010105981002M193)

摘　　要：目的研究α-细辛醚对难治性癫痫细胞模型的影响及对laminin β1表达的干预作用,探讨α-细辛醚抗癫痫的作用机制。方法体外培养海马神经元至第9天,随机将培养细胞分为正常对照组、模型组、模型+α-细辛醚(7.5μg/ml)组、模型+α-细辛醚(15μg/ml)组、模型+α-细辛醚(30μg/ml)组、模型+2μl无水乙醇组,观察各组神经元的形态学变化,检测各组细胞培养液乳酸脱氢酶(LDH)在12,24,48 h的变化。采用荧光定量PCR(FQ-PCR)和免疫组化检测各组laminin β1在处理后24 h的表达变化。结果①造模后少数神经元会发生迁移、融合,加α-细辛醚组均能减少神经细胞膜LDH的渗透。②在24 h,模型组lamininβ1 mRNA表达增加,各浓度α-细辛醚均能抑制laminin β1 mRNA的表达(均P<0.05)。③免疫组化显示laminin β1蛋白在细胞质和细胞外基质表达,平均光密度值:对照组(0.094 7±0.025 86),模型组(0.135 1±0.026 16),模型+30μg/mlα-细辛醚组(0.097 9±0.032 81)。结论α-细辛醚能减少难治性癫痫细胞模型神经元的细胞膜损伤,抑制laminin β1基因及蛋白的过度表达,这可能是α-细辛醚抗癫痫的作用机制之一。Objective To study the effects of α-asarone on the cell mode of intractable epilepsy and gene of laminin β1.Methods Cultured the hippocampal neurons in vitro for 9 days,and randomly divided into control group,model group,model＋α-asarone（7.5 μg/ml） group,model＋α-asarone（15 μg/ml） group,model＋α-asarone（30 μg/ml） group,model＋2 μl ethanol group.The morphological changes of neurons observed in different groups,and the lactate dehydrogenase（LDH） in cell supernatant was meatured at the time of 12,24,48 h.At 24 h,the FQ-PCR and immunohistochemical techniques were used to examine the expression of laminin β1.Results ①After the model was established,migration and fusion of neurons occured,and α-asarone could reduce the penetration of LDH from the membrane.At 24h,the expression of laminin β1 mRNA was increased in model group,but decreased after α-asarone was added,there was statistical difference between model group and model＋α-asarone group（P 0.05）.The immunohistochemical techniques showed the expression of laminin β1 protein at the cytoplasm and matrix.AOD： control group（0.094 7±0.025 9）,model group（0.135 1±0.026 2）,model＋α-asarone（30 μg/ml） group（0.097 9±0.032 8）.Conclusion α-asarone can reduce the membrane injury of neurons in the cell model of intractable epilepsy,and inhibit the excessive expression of laminin β1.It may be one way of α-asarone＇ s antiepileptic effect mechanisms.

关 键 词：Α-细辛醚 LAMININ β1 神经元 癫痫模型 

分 类 号：R742.1[医药卫生—神经病学与精神病学]