低频率电刺激通过调控Rho/ROCK/NF-κB途径对癫痫大鼠作用的研究

Effect of low-frequency stimulation on epileptic rats by regulating the Rho/ROCK/NF-κB pathway

摘要:

目的 探究低频率电刺激(LFS)对癫痫的作用及其可能的作用机制。方法 将60只SD大鼠随机分为对照组、癫痫模型组、假刺激组和LFS治疗组，每组15只。记录大鼠Racine 评分和脑电图，Morris水迷宫实验检测大鼠空间学习记忆能力；HE染色和尼氏染色检测大鼠海马CA1区病理学变化；TUNEL染色检测大鼠海马神经元细胞凋亡；ELISA检测大鼠血清TNF-α、IL-6和IL-1β水平；Western blotting检测大鼠海马RhoA、ROCK1、ROCK2、p65、p-p65和凋亡相关基因蛋白表达；RT-PCR检测大鼠海马RhoA、ROCK1和ROCK2 mRNA表达。结果 癫痫模型组大鼠出现了连续性棘波。癫痫模型组大鼠的Racine评分；神经元细胞凋亡率；Bax、c-caspase3和p-p65/p65蛋白表达；RhoA、ROCK1和ROCK2 mRNA和蛋白表达；血清TNF-α、IL-6和IL-1β水平较对照组升高，差异有统计学意义(P<0.05)。癫痫模型组大鼠的空间学习记忆能力、神经元数量、尼氏体数量、Bcl-2蛋白表达较对照组降低，差异有统计学意义(P<0.05)。LFS治疗组大鼠波峰下降。LFS治疗组大鼠的Racine评分；神经元细胞凋亡率；Bax、c-caspase3和p-p65/p65蛋白表达；RhoA、ROCK1和ROCK2 mRNA和蛋白表达；血清TNF-α、IL-6和IL-1β水平较假刺激组降低，差异有统计学意义(P<0.05)。LFS治疗组大鼠的空间学习记忆能力、神经元数量、尼氏体数量和Bcl-2蛋白表达较假刺激组升高，差异有统计学意义(P<0.05)。结论 LFS可能通过抑制Rho/ROCK/NF-κB途径来抑制炎症产生，从而发挥抗癫痫作用。

Abstract:

Objective To investigate the effect of low-frequency stimulation （LFS） on epilepsy and its possible mechanism.Methods A total of 60 Sprague-Dawley rats were randomly divided into control group （n=15）， epilepsy model group （n=15）， sham stimulation group （n=15）， and LFS treatment group （n=15）. The Racine scores and electroencephalograms of the rats were recorded. The Morris water maze test was used to test the spatial learning and memory ability of the rats； hematoxylin-eosin staining and Nissl staining were used to detect the pathological changes in the hippocampal CA1 region of the rats； TUNEL staining was used to determine the apoptosis of rat hippocampal neurons； enzyme-linked immunosorbent assay was used to measure the levels of TNF-α， IL-6， and IL-1β in rat serum； Western blot was used to determine the expression of RhoA， ROCK1， ROCK2， p65， p-p65， and apoptosis-related proteins in rat hippocampus； reverse transcription polymerase chain reaction was used to measure the expression of RhoA， ROCK1， and ROCK2 mRNA in rat hippocampus.Results Compared with the control group， the rats in the epilepsy model group showed continuous spike waves and had significantly higher Racine score， neuronal apoptosis rate， expression of Bax， c-caspase3， and p-p65/p65 proteins， as well as RhoA， ROCK1， and ROCK2 mRNA and proteins， and levels of TNF-α， IL-6， and IL-1β in serum （P<0.05）， and had significantly lower spatial learning and memory ability， number of neurons， number of Nissl bodies， and expression of Bcl-2 protein （P<0.05）. Compared with the sham stimulation group， the rats in the LFS treatment group had significantly lower wave crest， Racine score， neuronal apoptosis rate， expression of Bax， c-caspase3， and p-p65/p65 proteins， as well as RhoA， ROCK1， and ROCK2 mRNA and proteins， and levels of TNF-α， IL-6， and IL-1β in serum （P<0.05）， and had significantly higherspatial learning and memory ability， number of neurons， number of Nissl bodies， and expression of Bcl-2 protein （P<0.05）.Conclusions LFS may inhibit inflammation by inhibiting the Rho/ROCK/NF-κB pathway， thereby playing a protective role in epileptic rats.