School of Basic Medical Sciences, Hebei University of Chinese Medicine, Shijiazhuang, China; Department of Traditional Chinese Medicine, Anyang Vocational and Technical College, Anyang, China.
School of Basic Medical Sciences, Hebei University of Chinese Medicine, Shijiazhuang, China.
Brain Res. 2022 Jan 1;1774:147723. doi: 10.1016/j.brainres.2021.147723. Epub 2021 Nov 12.
Epilepsy is a common chronic neurological disorder worldwide, but its entire pathology remains unknown. The purpose of this study was to explore the antiepileptic effect of baicalin (BAL), the main bioactive component of scutellaria. We isolated astrocytes from neonatal rats and astrocytes were identified by glial fibrillary acidic protein (GFAP) immunostaining. The viability and phenotype of astrocytes were determined by Cell Counting Kit-8 (CCK-8) and immunofluorescence staining, respectively. For investigating the effect of BAL on the autophagy in A1 astrocytes treated PC12 cells, expression of light chain 3B (LC3-B) and sequestosome 1 (P62) was analyzed by immunofluorescence staining and apoptosis by acridine orange/ethidium bromide (AO/EB) staining, respectively. For animal experiments, pentylenetetrazol (PTZ)-induced epileptic model was used to explore the antiepileptic effect of BAL. The results showed that BAL reduced lipopolysaccharide (LPS)-induced complement C3 (C3, a marker of A1 astrocytes) + A1 cells and decreased autophagy and apoptosis in PC12 cells. Further findings showed seizure grade and latency were positively correlated with GFAP+/C3 + A1 cells' infiltration in interstitial astrocytes. After BAL treatment, epileptogenesis was ameliorated with decreased A1 astrocytes in the brain and improved behavioral performance. The enzyme-linked immunosorbent assay (ELISA) showed that the levels of interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α) were reduced in the cerebral interstitial site in the BAL group compared to the PTZ group. Western blotting analysis showed that BAL treatment reduced expression of C3, inward rectifier potassium channel Kir4.1, aquaporin-4 (AQP4) in the frontal cortex and Caspase-3, BCL2-associated X protein (Bax) in the hippocampus. In conclusion, these findings suggest that BAL can prevents cognitive and emotional disorders and has antiepileptic effects in rats, which may be associated with suppresses neuron autophagy and apoptosis in the hippocampus via regulate astrocyte phenotypes.
癫痫是一种常见的慢性神经系统疾病，全世界范围内都有，但它的全部病理仍然未知。本研究旨在探讨黄芩素（BAL）的抗癫痫作用，BAL 是黄芩的主要生物活性成分。我们从新生大鼠中分离出星形胶质细胞，并通过胶质纤维酸性蛋白（GFAP）免疫染色进行鉴定。通过细胞计数试剂盒-8（CCK-8）和免疫荧光染色分别确定星形胶质细胞的活力和表型。为了研究 BAL 对 PC12 细胞中 A1 星形胶质细胞自噬的影响，通过免疫荧光染色分析了微管相关蛋白轻链 3B（LC3-B）和自噬体相关蛋白 1（P62）的表达，通过吖啶橙/溴化乙锭（AO/EB）染色分析了细胞凋亡。对于动物实验，使用戊四氮（PTZ）诱导的癫痫模型来探索 BAL 的抗癫痫作用。结果表明，BAL 降低了脂多糖（LPS）诱导的补体 C3（C3，A1 星形胶质细胞的标志物）+A1 细胞，并减少了 PC12 细胞中的自噬和凋亡。进一步的研究结果表明，发作等级和潜伏期与间质星形胶质细胞中 GFAP+/C3+A1 细胞的浸润呈正相关。经过 BAL 治疗后，大脑中 A1 星形胶质细胞减少，癫痫发生得到改善，行为表现得到改善。酶联免疫吸附试验（ELISA）显示，与 PTZ 组相比，BAL 组大脑间质部位白细胞介素-1α（IL-1α）和肿瘤坏死因子-α（TNF-α）水平降低。Western blot 分析表明，BAL 治疗降低了额皮质中 C3、内向整流钾通道 Kir4.1、水通道蛋白-4（AQP4）和海马中 Caspase-3、B 细胞淋巴瘤-2 相关 X 蛋白（Bax）的表达。综上所述，这些发现表明，BAL 可以预防认知和情感障碍，并具有抗癫痫作用，这可能与通过调节星形胶质细胞表型抑制海马神经元自噬和凋亡有关。
