Given that the role of Gelsemine in neuroinflammation has been demonstrated, this research aimed to investigate the effect of Gelsemine on neonatal hypoxic-ischemic (HI) brain injury. An in vivo HI brain injury neonatal mouse model and an in vitro oxygen–glucose deprivation (OGD) cell model were established and pretreated with Gelsemine. The brain infarct volume, neuronal loss and apoptosis, as well as spatial learning and memory were examined by TTC staining, Nissl’s staining, TUNEL staining and Morris water maze test. Immunohistochemical staining was applied to detect the microglia cells and astrocytes in the mouse brain tissue. The cell viability was analyzed by CCK-8 assay. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), TNF-α, IL-1β, and IL-6 were determined via ELISA. The lactate dehydrogenase (LDH) release and reactive oxygen species (ROS) level in OGD-treated cells were detected by colorimetry and DCFH-DA staining. Nrf2, HO-1, and inflammation-related factors were analyzed by immunofluorescence, qRT-PCR, or western blot. Gelsemine reduced the infarct volume and neuronal loss and apoptosis, yet improved spatial learning and memory impairment of HI-injured mice. Gelsemine inhibited the elevated MDA, TNF-α, IL-1β, IL-6, LDH and ROS levels, promoted the reduced SOD level and viability, and strengthened the up-regulation of HO-1 and Nrf2 in brain tissues and OGD-treated cells. However, Nrf2 silencing reversed the effects of Gelsemine on the Nrf2/HO-1 pathway, inflammation, and oxidative stress in OGD-treated cells. Gelsemine produces neuroprotective effects on neonatal mice with HI brain injury by suppressing inflammation and oxidative stress via Nrf2/HO-1 pathway.

鉴于钩吻碱在神经炎症中的作用已得到证实，本研究旨在探讨钩吻碱对新生儿缺氧缺血 (HI) 脑损伤的影响。建立了体内 HI 脑损伤新生小鼠模型和体外氧-葡萄糖剥夺 (OGD) 细胞模型，并用钩吻碱预处理。通过TTC染色、尼氏染色、TUNEL染色和Morris水迷宫试验检测脑梗死体积、神经元丢失和凋亡以及空间学习和记忆能力。应用免疫组织化学染色检测小鼠脑组织中的小胶质细胞和星形胶质细胞。通过CCK-8测定法分析细胞活力。通过 ELISA 测定丙二醛 (MDA)、超氧化物歧化酶 (SOD)、TNF-α、IL-1β 和 IL-6 的水平。通过比色法和 DCFH-DA 染色检测 OGD 处理细胞中的乳酸脱氢酶 (LDH) 释放和活性氧 (ROS) 水平。通过免疫荧光、qRT-PCR 或蛋白质印迹分析 Nrf2、HO-1 和炎症相关因子。钩吻碱减少了梗塞体积和神经元丢失和细胞凋亡，但改善了 HI 损伤小鼠的空间学习和记忆障碍。钩吻碱抑制升高的 MDA、TNF-α、IL-1β、IL-6、LDH 和 ROS 水平，促进降低的 SOD 水平和活力，并加强脑组织和 OGD 处理的 HO-1 和 Nrf2 的上调细胞。然而，Nrf2 沉默逆转了钩吻碱对 OGD 处理细胞中 Nrf2/HO-1 通路、炎症和氧化应激的影响。