Liang Tianyu, Liu Renyang, Liu Jinquan, Hong Jun, Gong Fangxiao, Yang Xianghong
Emergency and Critical Care Center, Intensive Care Unit, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, 310014, Zhejiang, China.
Department of Critical Care Medicine, Zhejiang Provincial People's Hospital Bijie Hospital, Bijie, Guizhou, 551799, China.
Mol Neurobiol. 2025 Apr;62(4):4093-4114. doi: 10.1007/s12035-024-04534-5. Epub 2024 Oct 12.
Intracerebral hemorrhage (ICH) is an acute neurological disorder characterized by high mortality and disability rates. Previous studies have shown that 75% of patients who survive ICH experience varying degrees of neurological deficits. Sphk1 has been implicated in a multitude of phylogenetic processes, including innate immunity and cell proliferation. An in vivo rat model of ICH and an in vitro model of neuronal oxyhemoglobin (OxyHb) were constructed. The expression level of Sphk1 was assessed using western blotting and immunofluorescence, whereas cell death following ICH was evaluated using fluoro-Jade B and terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Immunofluorescence facilitated the examination of microglial phenotypic alterations, while enzyme-linked immunosorbent assays were used to determine the concentrations of inflammatory markers. Behavioral assays were employed to assess the overall behavioral modifications of animals. Neuronal Sphk1/Sirt1 protein levels gradually increased following the induction of ICH. Elevated Sphk1 expression resulted in increased levels of anti-inflammatory microglia and reduced levels of pro-inflammatory factors. In contrast, suppression of Sphk1 expression resulted in an increased number of dead cells, thereby exacerbating neurological deficits. In vitro findings indicated that the levels of phosphorylated PI3K and AKT proteins increased in conjunction with Sphk1 expression. This study established that after ICH, Sphk1 interacts with Sirt1 to mitigate neuroinflammation, cell death, oxidative stress, and brain edema via the PI3K/AKT signaling pathway. Augmenting expression of Sphk1 significantly can ameliorate neurological impairments induced by ICH, offering novel targets and perspectives for therapeutic interventions in ICH treatment.
脑出血(ICH)是一种急性神经疾病,其特征是死亡率和致残率高。先前的研究表明,脑出血幸存者中有75%会出现不同程度的神经功能缺损。鞘氨醇激酶1(Sphk1)参与了多种进化过程,包括先天免疫和细胞增殖。构建了脑出血的体内大鼠模型和神经元氧合血红蛋白(OxyHb)的体外模型。使用蛋白质免疫印迹法和免疫荧光法评估Sphk1的表达水平,而使用荧光玉髓B和末端脱氧核苷酸转移酶dUTP缺口末端标记染色评估脑出血后的细胞死亡情况。免疫荧光有助于检查小胶质细胞表型改变,同时使用酶联免疫吸附测定法测定炎症标志物的浓度。采用行为学检测来评估动物的整体行为变化。脑出血诱导后,神经元Sphk1/沉默调节蛋白1(Sirt1)蛋白水平逐渐升高。Sphk1表达升高导致抗炎小胶质细胞水平增加,促炎因子水平降低。相反,抑制Sphk1表达导致死亡细胞数量增加,从而加剧神经功能缺损。体外研究结果表明,磷酸化磷脂酰肌醇-3激酶(PI3K)和蛋白激酶B(AKT)蛋白水平随Sphk1表达而增加。本研究证实,脑出血后,Sphk1与Sirt1相互作用,通过PI3K/AKT信号通路减轻神经炎症、细胞死亡、氧化应激和脑水肿。显著增强Sphk1的表达可改善脑出血诱导的神经损伤,为脑出血治疗提供了新的治疗靶点和思路。