癫痫是最常见的神经系统疾病之一,但是这种疾病的神经生物学基础仍然知之甚少。对CBA小鼠脑(NeuN和caspase-8)的免疫表型与海马神经元在急性癫痫PTZ诱导的癫痫发作期间的功能状态和存活率评估是特别感兴趣的。这项研究的目的是研究NeuN和caspase-8在PTZ诱导的小鼠癫痫发作期间细胞周期调节和海马神经元死亡中的参与,并评估杨梅素在上述实验设置中的治疗效果。将雄性CBA小鼠(n=340)分为6组,以研究杨梅素和丙戊酸在PTZ诱导的癫痫发作模型中的神经保护和抗癫痫作用。第一组(对照,n=20)接受单次腹膜内注射0.9%NaCl溶液。第二组(仅限PTZ,n=110)接受单次腹膜内45mg/kgPTZ诱导癫痫发作。第三组(杨梅素+PTZ,n=90)以200mg/kg口服给予杨梅素5天,然后是PTZ注射。第四组(丙戊酸+PTZ,n=80)接受100mg/kg腹膜内丙戊酸5天,其次是PTZ。组V(杨梅素+NaCl,n=20)接受杨梅素和NaCl。VI组(丙戊酸+NaCl,n=20)接受丙戊酸和NaCl。使用改良的Racine量表监测癫痫发作的严重程度。行为评估包括感觉运动功能测试,使用旋转杆测试的电机协调,和认知功能通过莫里斯水迷宫。收集脑组织并分析氧化应激标志物,包括丙二醛(MDA),超氧化物歧化酶(SOD),和谷胱甘肽(GSH)。分析血液样本的细胞因子水平(IL-1β,IL-6和TNF-α)。组织学研究涉及H&E和Nissl染色以评估一般组织病理学和神经元密度。使用抗NeuN和caspase-8的抗体进行免疫组织化学分析以评估神经元细胞周期调节和凋亡。PTZ引起的癫痫发作引起显著的氧化应激和炎症,导致神经元损伤.生化分析显示MDA水平升高,SOD,GSH,IL-1β,IL-6和TNF-α。组织学和免疫组织化学评估显示,海马和其他大脑区域的caspase-8阳性神经元显着增加,NeuN阳性神经元减少,与癫痫发作严重程度相关。杨梅素和丙戊酸治疗可减少氧化应激标志物和神经元损伤。两种治疗都导致中度神经元保护,在海马中观察到更少的受损神经元,齿状回,和其他大脑区域与仅PTZ组相比。总结,杨梅素给药显示出有希望的神经保护作用。它显著降低了氧化应激标志物,包括MDA,并恢复抗氧化酶活性(SOD和GSH),表明了它的抗氧化潜力。杨梅素还能有效抑制促炎细胞因子IL-1β的升高,IL-6和TNF-α,表明强烈的抗炎特性。行为评估显示,杨梅素改善PTZ治疗小鼠的认知和运动功能,癫痫发作严重程度和死亡率显着降低。组织学分析支持这些行为发现,Nissl染色显示神经元损伤减少,NeuN染色显示杨梅素治疗组神经元完整性保存更好。此外,caspase-8染色提示神经元凋亡显著减少。
Epilepsy is one of the most frequently diagnosed neurological diseases, but the neurobiological basis of the disease remains poorly understood. Immunophenotyping CBA mice brain (
NeuN and caspase-8) in parallel with hippocampal neurons\' functional status and survival rate assessment during acute epileptic PTZ-induced seizures is of particular interest. The aims of this study were to investigate the involvement of
NeuN and caspase-8 in cell cycle regulation and the death of hippocampal neurons during PTZ-induced seizures in mice and to assess the therapeutic efficacy of Myricetin in the aforementioned experimental settings. Male CBA mice (n = 340) were divided into six groups to investigate the neuroprotective and antiepileptic effects of Myricetin and Valproic Acid in the PTZ-induced seizure model. Group I (control, n = 20) received a single intraperitoneal injection of NaCl 0.9% solution. Group II (PTZ only, n = 110) received a single intraperitoneal 45 mg/kg PTZ to induce seizures. Group III (Myricetin + PTZ, n = 90) was administered Myricetin orally at 200 mg/kg for 5 days, followed by a PTZ injection. Group IV (Valproic Acid + PTZ, n = 80) received intraperitoneal Valproic Acid at 100 mg/kg for 5 days, followed by PTZ. Group V (Myricetin + NaCl, n = 20) received Myricetin and NaCl. Group VI (Valproic Acid + NaCl, n = 20) received Valproic Acid and NaCl. Seizure severity was monitored using the modified Racine scale. Behavioral assessments included sensorimotor function tests, motor coordination using the rotarod test, and cognitive function via the Morris water maze. Brain tissues were collected and analyzed for oxidative stress markers, including malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH). Blood samples were analyzed for cytokine levels (IL-1β, IL-6, and TNF-α). Histological studies involved H&E and Nissl staining to evaluate general histopathology and neuronal density. Immunohistochemical analysis was conducted using antibodies against
NeuN and caspase-8 to assess neuronal cell cycle regulation and apoptosis. PTZ-induced seizures caused significant oxidative stress and inflammation, leading to neuronal damage. Biochemical analyses showed elevated levels of MDA, SOD, GSH, IL-1β, IL-6, and TNF-α. Histological and immunohistochemical evaluations revealed a significant increase in caspase-8-positive neurons and a decrease in
NeuN-positive neurons in the hippocampus and other brain regions, correlating with seizure severity. Myricetin and Valproic Acid treatments reduced oxidative stress markers and neuronal damage. Both treatments resulted in moderate neuronal protection, with fewer damaged neurons observed in the hippocampus, dentate gyrus, and other brain areas compared to the PTZ-only group. Summarizing, Myricetin administration showed promising neuroprotective effects. It significantly reduced oxidative stress markers, including MDA, and restored antioxidant enzyme activities (SOD and GSH), suggesting its antioxidative potential. Myricetin also effectively attenuated the elevation of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, indicating strong anti-inflammatory properties. Behavioral assessments revealed that Myricetin improved cognitive and motor functions in PTZ-treated mice, with notable reductions in seizure severity and mortality rates. Histological analyses supported these behavioral findings, with Nissl staining showing reduced neuronal damage and
NeuN staining indicating better preservation of neuronal integrity in Myricetin-treated groups. Additionally, caspase-8 staining suggested a significant reduction in neuronal apoptosis.