Feng Gang, Liu Xianghua, Wang Baoying, Li Ruixing, Chang Yaxin, Guo Nannan, Li Yawei, Chen Tiantian, Ma Bingxiang
College of Pediatric Medicine, Henan University of Chinese Medicine, Zhengzhou, 450046, China.
The First Affiliated Hospital of Henan University of Chinese Medicine, Shanghai, 450099, China.
Mol Biol Rep. 2023 Apr;50(4):3389-3399. doi: 10.1007/s11033-023-08301-y. Epub 2023 Feb 4.
The Chinese herbal formula Chaihujia Longgu Muli Decoction (CD) has a good antiepileptic effect, but its mechanisms remain unclear. Therefore, in this study we explored the molecular mechanisms of CD against epilepsy.
Twelve-day-old SD rats were randomly divided into a normal group, model group, valproic acid group, and CD high, medium, and low groups. Except for the normal group, the other groups were given an intraperitoneal injection of pentylenetetrazol (PTZ) to establish epilepsy models, and the Racine score was applied for model judgment. After 14 consecutive days of dosing, the Morris water maze test was performed. Then, hippocampal Nissl staining and immunofluorescence staining were performed, and synaptic ultrastructure was observed by transmission electron microscopy (TEM). RhoA/ROCK signaling pathway proteins were detected.
In PTZ model rats, the passing times were reduced, and the escape latency was prolonged in the Morris water maze test. Nissl staining showed that some hippocampal neurons swelled and ruptured, Nissl bodies in the cytoplasm were significantly reduced, and neurons were lost. Immunofluorescence detection revealed that the expression of PSD95 and SYP was significantly reduced. Electron microscopy results revealed that the number of synapses in hippocampal neurons was significantly reduced and the postsynaptic membrane length was significantly reduced. Western blot analysis showed that the RhoA/ROCK signaling pathway was activated, while SYP, SPD95, and PTEN expression was significantly decreased. After treatment with CD, neurobehavioral abnormalities and neuronal damage caused by epileptic seizures were improved.
CD exerted an antiepileptic effect by inhibiting the activation of the RhoA/ROCK signaling pathway.
中药柴胡加龙骨牡蛎汤（CD）具有良好的抗癫痫作用，但其机制尚不清楚。因此，在本研究中，我们探讨了CD抗癫痫的分子机制。
将12日龄的SD大鼠随机分为正常组、模型组、丙戊酸组以及CD高、中、低剂量组。除正常组外，其他组腹腔注射戊四氮（PTZ）建立癫痫模型，采用Racine评分进行模型判定。连续给药14天后，进行Morris水迷宫试验。然后，进行海马尼氏染色和免疫荧光染色，并通过透射电子显微镜（TEM）观察突触超微结构。检测RhoA/ROCK信号通路蛋白。
在PTZ模型大鼠中，Morris水迷宫试验中穿越次数减少，逃避潜伏期延长。尼氏染色显示，部分海马神经元肿胀破裂，细胞质中的尼氏体明显减少，神经元丢失。免疫荧光检测显示，PSD95和SYP的表达明显降低。电子显微镜结果显示，海马神经元中的突触数量明显减少，突触后膜长度明显缩短。蛋白质印迹分析表明，RhoA/ROCK信号通路被激活，而SYP、SPD95和PTEN的表达明显降低。CD治疗后，癫痫发作引起的神经行为异常和神经元损伤得到改善。
CD通过抑制RhoA/ROCK信号通路的激活发挥抗癫痫作用。
