Xiao Qihua, Tang Hai, Kong Lingwen, Ji Huiming, Liu Ya'nan, Cui Guiyun
Epilepsy Center, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China.
Rev Invest Clin. 2019;71(2):116-123. doi: 10.24875/RIC.18002650.
Adenosine A1 receptor (AA1R) is widely present in the central nervous system, exerting brain protective antiepileptic effects, mainly by binding corresponding G proteins. We evaluated the neuroprotective effects of AA1R on hippocampal neuronal injury after lithium chloride-pilocarpine-induced epilepsy in rats.
A total of 60 male SD rats were randomly divided into four groups (n = 15/group): normal control, epilepsy, epilepsy + AA1R antagonist (DPCPX), and epilepsy + AA1R agonist (2-CAdo). An epilepsy model was established through kindling by lithium chloride-pilocarpine. The four groups were observed on days 1, 14, and 30. Pathological and morphological changes of hippocampal neurons were observed by HE staining; apoptosis was detected by TUNEL assay. Caspase-3 and GABA receptor expressions were detected by Western blot.
In the hippocampal CA3 area of the epilepsy group, the cellular structure was not neatly arranged, and some neurons were swelling, thick, and incomplete. Compared with the epilepsy group at the same time point, cells in the epilepsy + DPCPX group had an increased distortion, disorganization, edema, cytoplasmic vacuoles, and degeneration. In the epilepsy + 2-CAdo group, cell arrangement was regular and orderly, and structural damages were lessened. Compared with the normal control group at the same time point, the epilepsy group underwent evident neuronal apoptosis, with a significantly higher apoptotic index (AI) (p < 0.05). Compared with the epilepsy group, the neuronal apoptosis of the epilepsy + DPCPX group was boosted, and the AI significantly increased (p < 0.05). The neuronal apoptosis of the epilepsy + 2-CAdo group was inhibited, and the AI significantly decreased (p < 0.05). Compared with the epilepsy group, the caspase-3 expression levels of the epilepsy + DPCPX group on days 14 and 30 were significantly upregulated (p < 0.05), but those of the epilepsy + 2-CAdo group were significantly downregulated (p < 0.05).
AA1R abated cell edema and reduced apoptosis, exerting neuroprotective effects on hippocampal neuronal injury after lithium chloride-pilocarpine-induced epilepsy.
腺苷A1受体（AA1R）广泛存在于中枢神经系统，主要通过与相应G蛋白结合发挥脑保护抗癫痫作用。我们评估了AA1R对氯化锂-匹罗卡品诱导的大鼠癫痫后海马神经元损伤的神经保护作用。
将60只雄性SD大鼠随机分为四组（每组n = 15）：正常对照组、癫痫组、癫痫 + AA1R拮抗剂（DPCPX）组和癫痫 + AA1R激动剂（2-CAdo）组。通过氯化锂-匹罗卡品点燃建立癫痫模型。在第1、14和30天对四组进行观察。通过HE染色观察海马神经元的病理和形态学变化；采用TUNEL法检测细胞凋亡。通过蛋白质免疫印迹法检测Caspase-3和GABA受体表达。
癫痫组海马CA3区细胞结构排列不整齐，部分神经元肿胀、增粗且不完整。与癫痫组同一时间点相比，癫痫 + DPCPX组细胞变形、紊乱、水肿、胞质空泡化及变性增加。癫痫 + 2-CAdo组细胞排列规则有序，结构损伤减轻。与正常对照组同一时间点相比，癫痫组神经元凋亡明显，凋亡指数（AI）显著升高（p < 0.05）。与癫痫组相比，癫痫 + DPCPX组神经元凋亡增加，AI显著升高（p < 0.05）。癫痫 + 2-CAdo组神经元凋亡受到抑制，AI显著降低（p < 0.05）。与癫痫组相比，癫痫 + DPCPX组在第14天和30天的Caspase-3表达水平显著上调（p < 0.05），而癫痫 + 2-CAdo组则显著下调（p < 0.05）。
AA1R减轻细胞水肿并减少凋亡，对氯化锂-匹罗卡品诱导的癫痫后海马神经元损伤发挥神经保护作用。
