Department of Anesthesiology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Shandong, China.
Department of Anesthesiology, Shanghai Deji Hospital Qingdao University, Shanghai, China.
Neurosci Lett. 2020 Jan 18;716:134647. doi: 10.1016/j.neulet.2019.134647. Epub 2019 Nov 22.
This study was to investigate the neuroprotective effect of erythropoietin (EPO) on hippocampal neuronal cell injury in developing rats.
The hippocampal neurons cells were obtained from SD rats aged 10 days and divided into control, propofol, EPO, and propofol + erythropoietin (E + P) groups. Cell proliferation and apoptosis were measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Ki-67 immunofluorescence, and flow cytometry, respectively. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-4 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA). Cellular immunohistochemistry was utilized to detect the expression of proliferating cell nuclear antigen (PCNA), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3). Quantitative real time polymerase chain reaction (qRT-PCR) and western blot were used to detect the expression of Bax, Bcl-2, Caspase-3, toll-like receptor 4 (TLR4) and p65. Furthermore, TLR4 antagonist (TAK-242) and activator (LPS) were used to study the relationship between EPO and TLR4.
Propofol treatment caused morphological and structural damage of hippocampal neurons. However, EPO significantly improved this damage, enhanced cell proliferation, decreased apoptosis and pro-inflammatory factor content, up-regulated the expression of Ki-67, PCNA, Bcl-2, NGF, BDNF and NT-3, as well as decreased the expression of Bax, Caspase-3, TLR4 and p65 (p < 0.05). After TAK-242 or LPS treatment, it showed similar results in propofol + TAK-242 (T + P) group and E + P group.
Erythropoietin could attenuate propofol-induced hippocampal neuronal cell injury in developing rats, which may be related to inhibit TLR4 expression.
本研究旨在探讨促红细胞生成素（EPO）对发育期大鼠海马神经元细胞损伤的神经保护作用。
取 10 日龄 SD 大鼠海马神经元细胞，分为对照组、丙泊酚组、EPO 组和丙泊酚+EPO（E+P）组。分别采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、Ki-67 免疫荧光、流式细胞术检测细胞增殖和凋亡；酶联免疫吸附试验（ELISA）检测肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-6、IL-4 和 IL-10 水平；细胞免疫组化法检测增殖细胞核抗原（PCNA）、神经生长因子（NGF）、脑源性神经营养因子（BDNF）和神经营养因子-3（NT-3）的表达；实时荧光定量聚合酶链反应（qRT-PCR）和 Western blot 检测 Bax、Bcl-2、Caspase-3、Toll 样受体 4（TLR4）和 p65 的表达。此外，还使用 TLR4 拮抗剂（TAK-242）和激动剂（LPS）研究 EPO 与 TLR4 之间的关系。
丙泊酚处理导致海马神经元形态和结构损伤，而 EPO 则明显改善了这种损伤，增强了细胞增殖，降低了细胞凋亡和促炎因子含量，上调了 Ki-67、PCNA、Bcl-2、NGF、BDNF 和 NT-3 的表达，下调了 Bax、Caspase-3、TLR4 和 p65 的表达（p<0.05）。在使用 TAK-242 或 LPS 处理后，丙泊酚+TAK-242（T+P）组和 E+P 组的结果类似。
EPO 可减轻发育期大鼠丙泊酚诱导的海马神经元细胞损伤，这可能与抑制 TLR4 表达有关。
