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柴胡加龙骨牡蛎汤对难治性癫痫大鼠神经元损伤的影响及机制
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| 篇名： | 柴胡加龙骨牡蛎汤对难治性癫痫大鼠神经元损伤的影响及机制 |
| TITLE： | Effects of Chaihu longgu muli decoction on neuronal damage in refractory epilepsy rats and its mechanism |
| 摘要： | 目的 探讨柴胡加龙骨牡蛎汤对难治性癫痫大鼠海马神经元损伤的影响及潜在作用机制。方法采用侧脑室注射海人藻酸建立难治性癫痫大鼠模型，将难治性癫痫大鼠随机分为模型组、塞来昔布组（阳性对照）、柴胡加龙骨牡蛎汤组、柴胡加龙骨牡蛎汤+空载组、柴胡加龙骨牡蛎汤+环氧合酶-2（COX-2）过表达组，每组12只；另取12只大鼠为假手术组（侧脑室注射生理盐水）。各组给予相应的药物/生理盐水8周后，采用Racine分级标准评估大鼠癫痫发作行为学，观察海马组织CA3区病理学变化和细胞凋亡情况；采用酶联免疫吸附测定法检测海马组织COX-2、前列腺素E2（PGE2）、肿瘤坏死因子α（TNF-α）水平；采用比色法检测海马组织丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性；分别采用实时荧光定量聚合酶链反应和Westernblot法检测海马组织COX-2、P糖蛋白（P-gp）mRNA和蛋白的表达。结果假手术组大鼠无癫痫发作；与假手术组比较，模型组大鼠癫痫Racine分级，海马组织细胞凋亡率，COX-2、PGE2、TNF-α水平和MDA含量以及COX-2、P-gp的mRNA和蛋白相对表达水平均显著升高，SOD活性显著降低（P＜0.05），海马CA3区神经元出现明显损伤；与模型组比较，塞来昔布组、柴胡加龙骨牡蛎汤组大鼠癫痫Racine分级，海马组织细胞凋亡率，COX-2、PGE2、TNF-α水平和MDA含量以及COX-2、P-gp的mRNA和蛋白相对表达水平均显著降低，SOD活性均显著升高（P＜0.05），海马CA3区神经元损伤减轻；与柴胡加龙骨牡蛎汤组、柴胡加龙骨牡蛎汤+空载组比较，柴胡加龙骨牡蛎汤+COX-2过表达组大鼠上述指标的改善被显著逆转（P＜0.05），海马组织CA3区神经元损伤加重。结论柴胡加龙骨牡蛎汤可能通过抑制COX-2/PGE2信号通路的激活，抑制炎症和氧化应激，从而减轻难治性癫痫大鼠海马神经元损伤。 |
| ABSTRACT： | OBJECTIVE To explore the effect and potential mechanism of Chaihu longgu muli decoction on hippocampal neuronal damage in refractory epilepsy rats. METHODS The refractory epilepsy rat model was established by intracerebroventricular injection of kainic acid. The refractory epilepsy rats were randomly assigned into model group， celecoxib group （positive control）， Chaihu longgu muli decoction group， Chaihu longgu muli decoction+empty loading group， Chaihu longgu muli decoction+cyclooxygenase-2 （COX-2） overexpression group， with 12 rats in each group. Another 12 rats were selected as sham operation group （injected with normal saline into the lateral ventricle）. After 8 weeks of corresponding drug/normal saline intervention， Racine grading criteria were applied to evaluate the seizure behavior of rats； pathological changes and cell apoptosis in CA3 region of hippocampal tissue were observed； ELISA method was adopted to detect the levels of COX-2， prostaglandin E2 （PGE2）， and tumor necrosis factor-α （TNF-α） in hippocampal tissue. The colorimetric method was applied to detect the content of malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） in hippocampal tissue. Real-time fluorescence quantitative PCR and Western blot were applied to detect the mRNA and protein expressions of COX-2 and P-glycoprotein （P-gp） in hippocampal tissue. RESULTS The rats in the sham operation group had no epileptic seizures. Compared with sham operation group， Racine grade of epilepsy， apoptotic rate of hippocampal tissue cells， COX-2， PGE2 and TNF-α levels， MDA contents， mRNA and protein levels of COX-2 and P-gp in the model group increased significantly， while SOD activity decreased significantly （P＜0.05）； prominent damage was observed in the hippocampal CA3 region. Compared with model group， Racine grade of epilepsy， apoptotic rate of hippocampal tissue cells， COX-2， PGE2 and TNF-α levels， MDA contents， mRNA and protein levels of COX-2 and P-gp in celecoxib group and Chaihu longgu muli decoction group decreased significantly， while SOD activity increased significantly（P＜0.05）； the neuronal damage in hippocampal CA3 region was alleviated. Compared with Chaihu longgu muli decoction group and Chaihu longgu muli decoction+ empty loading group， the improvement in the aforementioned indicators in rats from Chaihu longgu muli decoction+COX-2 overexpression group was significantly reversed （P＜0.05）， with exacerbated neuronal damage in the CA3 region of the hippocampal tissue. CONCLUSIONS Chaihu longgu muli decoction may alleviate hippocampal neuronal damage in refractory epilepsy rats by inhibiting the activation of COX-2/PGE2 signaling pathway， inhibiting inflammation and oxidative stress， thereby alleviating hippocampal neuronal damage in refractory epilepsy rats. |
| 期刊： | 2025年第36卷第06期 |
| 作者： | 单萍；张继龙 |
| AUTHORS： | SHAN Ping，ZHANG Jilong |
| 关键字： | 难治性癫痫；柴胡加龙骨牡蛎汤；神经元损伤；环氧合酶-2/前列腺素E2信号通路 |
| KEYWORDS： | refractory epilepsy； Chaihu longgu muli decoction； neuronal damage； COX-2/PGE2 signaling pathway |
| 阅读数： | 3 次 |
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