Department of Neurology, The Second Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang, 314000, China.
Department of Neurology, The Second Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang, 314000, China.
Epilepsy Res. 2019 Aug;154:90-96. doi: 10.1016/j.eplepsyres.2019.05.007. Epub 2019 May 13.
Neuronal apoptosis is a regulated intrinsic cell mechanism and common pathological phenomenon after seizures, which involves the protein kinase B/cAMP response element binding protein / brain derived neurotrophic factor (AKT/CREB/ BDNF) signaling pathway. In this study, we aimed to identify the effects of salvianolic acid B (Sal B), a major water-soluble component of the Chinese herb, Danshen, on rats in which seizures had been induced by pentylenetetrazole (PTZ) and the underlying molecular mechanisms mediating these effects. For this, 60 adult male Sprague-Dawley rats were divided into a control group, a 'PTZ' group and a 'PTZ + Sal B' group. The animals in the control group received an intraperitoneal (i.p.) injection of saline on alternate days for a total of 15 injections and saline orally once a day for 29 days. The animals in the 'PTZ' group received PTZ (40 mg/kg, i.p.) on alternate days for a total of 15 injections and saline orally once a day for 29 days. Similarly, the animals in the 'PTZ + Sal B' group received PTZ (40 mg/kg, i.p.) on alternate days and Sal B (20 mg/kg) orally once a day for 29 days. Neural density was then evaluated using immunofluorescence (IF) staining of microtubule-associated protein 2 (MAP2). Neuronal apoptosis was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining. In addition, the expression of several proteins related to AKT/CREB/BDNF signaling was measured using Western blotting. The results indicated that more severe seizures, decreased neural density, decreased expression of Bcl-2, increased expression of Bax and cleaved caspase-3, and inactivation of AKT/CREB/BDNF signaling occurred in the 'PTZ' group in comparison with the control group. However, those changes were suppressed by Sal B. Thus, these data suggest that Sal B has anticonvulsant and anti-apoptotic effects in a PTZ-induced seizure model through activation of the AKT/CREB/BDNF signaling pathways.
神经元凋亡是一种受调控的内在细胞机制，也是癫痫发作后的常见病理现象，涉及蛋白激酶 B/cAMP 反应元件结合蛋白/脑源性神经营养因子（AKT/CREB/BDNF）信号通路。本研究旨在探讨丹参水溶性成分丹参酸 B（Sal B）对戊四氮（PTZ）诱导的癫痫大鼠的作用及其潜在的分子机制。为此，将 60 只成年雄性 Sprague-Dawley 大鼠分为对照组、PTZ 组和 PTZ+Sal B 组。对照组大鼠隔日腹腔注射生理盐水，共 15 次，每日口服生理盐水 1 次，共 29 天；PTZ 组大鼠隔日腹腔注射 PTZ（40mg/kg），共 15 次，每日口服生理盐水 1 次，共 29 天；PTZ+Sal B 组大鼠隔日腹腔注射 PTZ（40mg/kg），每日口服 Sal B（20mg/kg），共 29 天。然后采用免疫荧光（IF）染色法检测微管相关蛋白 2（MAP2）评估神经密度，末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）染色检测神经元凋亡，Western blot 法检测 AKT/CREB/BDNF 信号相关蛋白的表达。结果表明，与对照组相比，PTZ 组大鼠癫痫发作更严重，神经密度降低，Bcl-2 表达减少，Bax 和 cleaved caspase-3 表达增加，AKT/CREB/BDNF 信号通路失活。然而，Sal B 可抑制这些变化。因此，这些数据表明，Sal B 通过激活 AKT/CREB/BDNF 信号通路，在 PTZ 诱导的癫痫模型中具有抗惊厥和抗凋亡作用。
