熄风胶囊对难治性癫痫大鼠海马Ⅰ型钠通道α亚基蛋白及mRNA表达的影响-《西部中医药》

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Title:: Effects of XiFeng Capsules on α Subunit Protein and mRNA Expressions of Type I Sodium Channel in the Hippocampi of the Rats Suffering Intractable Epilepsy

文章编号:: 1004-6852(2018)04-0014-06

作者:: 房艳艳1，李新民2△，路岩莉2，韩耀巍2，聂坤2，吴超1; 1 天津中医药大学，天津 300193； 2 天津中医药大学第一附属医院

Author(s):: FANG Yanyan1, LI Xinmin2△, LU Yanli2, HAN Yaowei2, NIE Kun2, WU Chao1; 1 Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China 2 First Teaching Hospital of Tianjin University of TCM

关键词:: 难治性癫痫; SCN1A; 熄风胶囊; Ⅰ型钠通道; α亚基蛋白

Keywords:: intractable epilepsy; SCN1A; XiFeng capsules; type I sodium channel; α subunit protein

分类号:: R742.1

文献标志码:: A

摘要:: 目的：探讨熄风胶囊对氯化锂—匹罗卡品致难治性癫痫模型大鼠Ⅰ型钠通道α亚基蛋白及mRNA表达的影响。方法：建立氯化锂—匹罗卡品癫痫大鼠模型。实验大鼠随机分为8组：正常对照组(空白组)、模型对照组(模型组)、熄风胶囊低剂量组(熄低组)、熄风胶囊中剂量组(熄中组)、熄风胶囊高剂量组(熄高组)、卡马西平治疗组(CBZ组)、熄风胶囊中剂量+卡马西平组(熄卡组)、熄风胶囊中剂量+1/2卡马西平组(熄卡低组)。熄低、中、高组分别予熄风胶囊0.33 g、0.66 g、0.99 g，浓缩剂2 mL；CBZ组予CBZ20 mg/kg；熄卡、熄卡低组分别予熄风胶囊0.66 g和CBZ 20 mg/kg、CBZ 10 mg/kg；模型组和空白组分别予生理盐水2 mL。每天上午灌胃1次，共持续60天。给药结束后检测各组大鼠Ⅰ型钠通道α亚基蛋白及mRNA的表达。结果：1)免疫组化染色法示：与空白组比较，模型组SCN1A的表达高于空白组(P＜0.05)；与模型组比较，各组治疗SCN1A的表达均低于模型组(P＜0.05)；与CBZ组相比，熄卡组SCN1A的表达均低于CBZ组(P＜0.05)；2)Western-blot示：与空白组比较，模型组SCN1A的表达高于空白组(P＜0.05)；与模型组比较，各治疗组SCN1A的表达均低于模型组(P＜0.05)；与CBZ 组比较，熄卡低组SCN1A的表达低于CBZ组(P＜0.05)；3)Real-time RT-PCR示：熄高组、熄中组、熄低组、熄卡组对 SCN1A mRNA 表达有抑制作用，而卡马组、熄卡低组对SCN1A mRNA 表达有促进作用。结论：免疫组化、Western-blot、Real-timeRT-PCR 结果显示：与正常大鼠相比，IE大鼠海马SCN1A在蛋白与mRNA两个层面均表达上调。熄风胶囊可能通过抑制癫痫大鼠海马Ⅰ型钠通道α亚基蛋白及基因的表达抑制钠电流，从而降低癫痫反复自发性发作的可能。

Abstract:: Objective: To discuss the effects of XiFeng capsules on α subunit protein and mRNA expressions of type I sodium channel in the hippocampi of the epileptic rats induced by lithium chloride-pilocarpine. Methods:Lithium chloride-pilocarpine induced epileptic rat model was established. The rats were randomized into eight groups: normal control group (the blank group), model control group (the model group), low, moderate and high dose groups of XiFeng capsules (low XF group, moderate XF group and high XF group), the treatment group of carbamazepine (CBZ group), moderate dose group of XiFeng capsules and carbamazepine (XF+CBZ group) as well as moderate dose group of XiFeng capsules and 1/2 carbamazepine (XF+ low CBZ group), the groups of low XF, moderate XF and high XF were given XiFeng capsules respectively, 0.33 g, 0.66 g and 0.99 g, inspissant 2 mL; CBZ group accepted CBZ 20 mg/kg; XF+CBZ group and XF+ low CBZ group received XiFeng capsules, 0.66 g, and CBZ 20 mg/kg, CBZ 10 mg/kg; the model group and the blank group were given physiological saline respectively,2 mL, lavage once in the morning each day, for 60 days consecutively. The expressions of mRNA and α subunit protein of type I sodium channel of the rats in different groups were detected after the administration. Results:1) Immunohistochemical staining showed: compared with the blank group, the expressions of SCN1A in the model group were higher than these of the blank group (P＜0.05); compared with the model group, the groups of low XF, moderate XF and high XF, XF+CBZ group and XF+ low CBZ group were lower than the model group in the expressions of SCN1A (P＜0.05); when different treatment groups were compared with CBZ group, the groups of low XF, moderate XF and high XF were lower than CBZ group in the expressions of SCN1A (P＜0.05); 2) Westernblot demonstrated: compared with the blank group, the model group was higher than the blank group in the expressions of SCN1A(P＜0.05), the group of high XF, XF+CBZ group and XF+ low CBZ group were lower than the blank group in the expressions of SCN1A(P＜0.05); compared with the model group, different treatment groups were lower than the model group in the expressions of SCN1A (P＜0.05); compared with CBZ group, XF+CBZ group and XF+ low CBZ group were lower than CBZ group in the expressions of SCN1A(P＜0.05); 3)Real-time RT-PCR showed: the groups of low XF, moderate XF and high XF, and XF+CBZ group inhibited the expressions of SCN1A mRNA, while CBZ group and XF+ low CBZ group could promote the expressions of SCN1A mRNA. Conclusion: The results of immunohistochemical staining, western-blot and Real-time RT-PCR showed: compared with normal rats, the expressions of SCN1A protein and mRNA in the hippocampi of IE rats rose. XiFeng capsules could reduce the possibility of recurrent spontaneous attack of epilepsy, which might be realized by inhibiting sodium current, α subunit protein and mRNA expressions of type I sodium channel in the hippocampi of the epileptic rats.

《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

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