MiRNA和miR-146a在托吡酯治疗颞叶癫痫的临床意义
发布时间：2018-04-05 18:31
本文选题：颞叶癫痫 切入点：miR-146a 出处：《延边大学》2014年博士论文
【摘要】：研究背景：癫痫是一种常见的神经失调症,并以反复自发异常的神经元放电为主要特点,目前抗癫痫药物的使用是治疗颞叶癫痫最重要的手段,托吡酯(TPM)则是其中最有效的抗癫痫药物之一,然而TPM治疗癫痫的具体机制尚未明确。MicroRNAs (miRNAs)一种非编码单链小分子RNA,近几年的研究表明它不仅对神经发育产生影响,还有研究表明它会参与到癫痫的发生发展中,但是其具体机制尚不明确。结合国内外的许多研究并未发现miRNAs与TPM治疗癫痫机制的相关论文,这为明确癫痫发病机制和TPM治疗癫痫的机制提供了基础。本实验旨在探讨microRNAs (miRNAs)在颞叶癫痫中的表达谱改变,并通过锂-匹罗卡品颞叶癫痫大鼠模型制备,阐述TPM对于癫痫大鼠海马凋亡的影响和机制。 材料和方法：将大鼠随机分为四组：1)正常对照组,2)颞叶癫痫组,3)TPM低剂量组(40mg/kg),4) TPM高剂量组(80mg/kg)。通过基因芯片分析技术测定miRNAs在上述四组中的表达谱改变,HE染色观察各组大鼠海马组织与细胞的形态学改变,并利用免疫蛋白印迹,免疫组化以及免疫荧光等方法测定上述各组海马中caspase-8蛋白的表达水平。 结果：与对照组相比,TLE大鼠海马中检测到了12个上调的miRNAs (miR-146a,-183,-204,-210,-339,-34b,351,-429,-455,466b,-503, and-532)和15个下调的miRNAs(miR-34a,-383,-451,-101a,-488,-499,-300,-551b,-380,-592,-598,-741,-7b,-374, and-9)。在使用TPM治疗后,12个上调的miRNAs发生了下调,15个下调的miRNAs则出现了上调。 miR-146a是上调的miRNAs中变化最明显的。这一结果阐明,TPM能够通过主要下调 miR-146a的表达治疗癫痫。另外,在TLE大鼠海马中可以检测到caspase-8的表达水平明 显增加,但是经TPM治疗后,其表达水平下降,证实了TPM能够通过下调caspase-8的 表达抑制TLE大鼠海马细胞的凋亡,同时还发现,低剂量TPM处理组中大鼠海马caspase-8 的表达较高剂量TPM处理组更低,表明低剂量TPM的对大鼠癫痫的疗效比高剂量TPM 更好。通过对大鼠海马组织上述各组的免疫组化、免疫荧光和HE染色结果分析发现, 癫痫大鼠的海马中DG区的损伤最为严重,其次为CA3区,但是经过治疗后,其死亡的神 经元细胞明显减少,证实了TPM的疗效显著,且TPM低剂量的效果较TPM高剂量更为 明显,这与免疫蛋白印迹的结果一致。结论：1)TPM可以通过下调caspase-8的表达抑制癫痫小鼠海马细胞凋亡的发生； 2)在TLE的发生发展过程中存在miRNAs特异表达谱,其中miR-146a在TPM治疗TLE 过程中具有明显作用,有望成为TLE治疗的分子靶点。
[Abstract]:Background: epilepsy is a common neurological disorder characterized by recurrent spontaneous discharges of neurons. At present the use of antiepileptic drugs is the most important method for the treatment of temporal lobe epilepsy.Topiramate (TPM) is one of the most effective antiepileptic drugs. However, the specific mechanism of TPM in the treatment of epilepsy is not clear.Other studies have shown that it may be involved in the development of epilepsy, but its mechanism is unclear.In combination with many studies at home and abroad, the related papers on the mechanism of miRNAs and TPM in the treatment of epilepsy have not been found, which provides a basis for clarifying the pathogenesis of epilepsy and the mechanism of TPM in the treatment of epilepsy.The purpose of this study was to investigate the changes of microRNAs miRNAs) expression profile in temporal lobe epilepsy and to elucidate the effect and mechanism of TPM on hippocampal apoptosis in rats with lithium-pilocarpine temporal lobe epilepsy.Materials and methods: rats were randomly divided into four groups: 1) normal control group (2)) temporal lobe epilepsy group (30 mg 路kg ~ (-1)) low dose group (40 mg / kg ~ (4)) TPM high dose group (n = 80 mg / kg ~ (-1)).The changes of miRNAs expression profile in the above four groups were detected by gene chip analysis technique. The morphological changes of hippocampus and cells in each group were observed by HE staining, and Western blot was used.Immunohistochemical and immunofluorescence methods were used to detect the expression of caspase-8 protein in hippocampus.缁撴灉:涓庡鐓х粍鐩告瘮,TLE澶ч紶娴烽┈涓娴嬪埌浜，
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