拉莫三嗪与丙戊酸钠对Ⅰ型钠离子通道Na_v1.1膜蛋白表达的影响
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摘要
目的比较Ⅰ型钠离子通道NaV1.1在拉莫三嗪(LTG)及丙戊酸钠(VPA)处理后的蛋白表达水平,探讨LTG和VPA参与癫痫治疗的可能机制。方法构建带YFP标签的Na_V1.1蛋白表达载体(pCMV-YFP-SCN1A),转染HEK293T细胞。使用LTG、 VPA处理24 h,提取总蛋白和生物素标记的膜蛋白,采用Western blotting检测Na_V1.1总蛋白及膜蛋白的表达水平。结果与对照组比较, Na_V1.1膜蛋白表达水平在VPA处理24 h后显著增加(P <0.05),而在LTG处理24 h后无显著改变(P>0.05); Na_V1.1总蛋白表达水平在LTG、 VPA处理24 h后无明显变化(P>0.05)。结论 VPA可促进Na_V1.1膜蛋白表达,可能是参与抗癫痫治疗过程的潜在机制。
Objective To compare the expression of lamotrigine(LTG) and valproate(VPA) on Nav1.1 protein, and to explore the possible mechanism of LTG and VPA involved in the treatment of epilepsy. Methods The pCMV-YFP-SCN1 A plasmid was constructed and used to transfect HEK293 T cells. The transfected HEK293 T cells were treated with LTG or VPA for 24 hours, respectively. The total protein and biotin-labeled membrane protein were extracted, and Western blotting were used to detect the expression of Nav1.1 total protein and membrane protein. Results Compared with the control group, the expression of Nav1.1 membrane protein significantly increased with24-hour VPA treatment(P <0.05), but no significant change with 24-hour LTG treatment(P >0.05). No significant difference was found in the expression of Nav1.1 total protein with 24-hour VPA or LTG treatment(P >0.05). Conclusions VPA can promote the expression of Nav1.1 membrane protein, which may be a potential mechanism that involved in anti-epileptic therapy.
Objective To compare the expression of lamotrigine(LTG) and valproate(VPA) on Nav1.1 protein, and to explore the possible mechanism of LTG and VPA involved in the treatment of epilepsy. Methods The pCMV-YFP-SCN1 A plasmid was constructed and used to transfect HEK293 T cells. The transfected HEK293 T cells were treated with LTG or VPA for 24 hours, respectively. The total protein and biotin-labeled membrane protein were extracted, and Western blotting were used to detect the expression of Nav1.1 total protein and membrane protein. Results Compared with the control group, the expression of Nav1.1 membrane protein significantly increased with24-hour VPA treatment(P <0.05), but no significant change with 24-hour LTG treatment(P >0.05). No significant difference was found in the expression of Nav1.1 total protein with 24-hour VPA or LTG treatment(P >0.05). Conclusions VPA can promote the expression of Nav1.1 membrane protein, which may be a potential mechanism that involved in anti-epileptic therapy.
引文
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[12] Rusconi R, Combi R, Cestèle S, et al. A rescuable folding defective Nav1.1(SCN1A)sodium channel mutant causes GEFS+:common mechanism in Nav1.1 related epilepsies?[J]. Hum Mutat, 2009, 30(7):E747-760.
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[14] Thompson CH, Porter JC, Kahlig KM, et al. Nontruncating SCN1A mutations associated with severe myoclonic epilepsy of infancy impair cell surface expression[J]. Biol Chem, 2012, 287(50):42001-42008.
[15] Misra SN, Kahlig KM, George AL Jr. Impaired NaV1.2 function and reduced cell surface expression in benign familial neonatal-infantile seizures[J]. Epilepsia, 2008, 49(9):1535-1545.
[16] Yu FH, Mantegazza M, Westenbroek RE, et al. Reduced sodium current in GABAergic interneurons in a mouse model of severe myoclonic epilepsy in infancy[J]. Nat Neurosci, 2006, 9(9):1142-1149.
[2] Meng H, Xu HQ, Yu L, et al. The SCN1A mutation database:updating information and analysis of the relationships among genotype, functional alteration, and phenotype[J]. Hum Mutat, 2015, 36(6):573-580.
[3] Scheffer IE, Berkovic SF. Generalized epilepsy with febrile seizures plus. A genetic disorder with heterogeneous clinical phenotypes[J].Brain, 1997, 120(3):479-490.
[4] Ohmori I, Ouchida M, Ohtsuka Y, et al. Significant correlation of the SCN1A mutations and severe myoclonic epilepsy in infancy[J].Biochem Biophys Res Commun, 2002, 295(1):17-23.
[5]陈秀敏.拉莫三嗪和丙戊酸钠联合治疗儿童难治性癫痫临床观察[J].中国病案, 2011, 12(3):63-64.
[6]陈秋文.拉莫三嗪联合丙戊酸钠治疗儿童难治性癫痫临床观察[J].中山大学学报(医学科学版), 2009, 30(z2):113-115.
[7] Perucca E. An introduction to antiepileptic drugs[J]. Epilepsia,2005, 46(S4):31-37.
[8] Macdonald RL, Kelly KM. Antiepileptic drug mechanisms of action[J]. Epilepsia, 1993, 34(S5):1-8.
[9] Weintraub D, Buchsbaum R, Resor SR Jr, et al. Effect of antiepileptic drug comedication on lamotrigine clearance[J]. Arch Neurol, 2005,62(9):1432-1436.
[10] Stefani A, Spadoni F, Siniscalchi A, et al. Lamotrigine inhibits Ca2+currents in cortical neurons:functional implications[J]. Eur J Pharmacol, 2006, 307(1):113-116.
[11] Abdelsayed M, Sokolov S. Voltage-gated sodium channels:pharmaceutical targets via anticonvulsants to treat epileptic syndromes[J].Channels(Austin), 2013, 7(3):146-152.
[12] Rusconi R, Combi R, Cestèle S, et al. A rescuable folding defective Nav1.1(SCN1A)sodium channel mutant causes GEFS+:common mechanism in Nav1.1 related epilepsies?[J]. Hum Mutat, 2009, 30(7):E747-760.
[13] Rusconi R, Scalmani P, Cassulini RR, et al. Modulatory proteins can rescue a trafficking defective epileptogenic Nav1.1 Na+channel mutant[J]. J Neurosci, 2007, 27(41):11037-11046.
[14] Thompson CH, Porter JC, Kahlig KM, et al. Nontruncating SCN1A mutations associated with severe myoclonic epilepsy of infancy impair cell surface expression[J]. Biol Chem, 2012, 287(50):42001-42008.
[15] Misra SN, Kahlig KM, George AL Jr. Impaired NaV1.2 function and reduced cell surface expression in benign familial neonatal-infantile seizures[J]. Epilepsia, 2008, 49(9):1535-1545.
[16] Yu FH, Mantegazza M, Westenbroek RE, et al. Reduced sodium current in GABAergic interneurons in a mouse model of severe myoclonic epilepsy in infancy[J]. Nat Neurosci, 2006, 9(9):1142-1149.