Yan Ke, Bian Jiarong, He Liang, Song Bingwei, Shen Linhai, Zhen Yong
Department of Neurosurgery, Northern Jiangsu People's Hospital Affiliated to Yangzhou University, Yangzhou 225001, PR China.
Department of Respiratory Medicine, Northern Jiangsu People's Hospital Affiliated to Yangzhou University, Yangzhou 225001, PR China.
Mol Immunol. 2025 Mar;179:18-28. doi: 10.1016/j.molimm.2025.01.002. Epub 2025 Feb 5.
We aimed to explore the role of SIRT3 in ameliorating neuroinflammation caused by hypoxia-ischemia (HI).
A rat model of HI was established, and 48 hours prior to constructing the HI model, the rats received an intracerebroventricular injection of a recombinant adeno-associated virus type 9 vector. TTC and Nissl staining assessed the effects of SIRT3 on cerebral infarction and brain atrophy in HI rats. Neuroinflammation was evaluated by investigating IL-1β and MPO positive cells, and ELISA for determining inflammatory cytokines. IBA-1, CD68, and iNOS positive microglia and NLRP3 activation-related protein expression were also detected. SIRT3 was overexpressed in oxygen glucose deprivation (OGD)-induced microglia model, where cell morphology and expressions of pro-inflammatory cytokines and NLRP3 inflammasome activation-related proteins were examined. Additionally, neurons co-cultured with SIRT3-overexpressing microglia were analyzed for mitochondrial damage and apoptosis.
SIRT3 alleviated cerebral infarction and atrophy in HI rats. It also inhibited neuroinflammation, reducing IL-1β and MPO positive cells, and lowered the levels of pro-inflammatory cytokines. In both HI rat model and OGD cell model, SIRT3 inhibited excessive activation of microglia and NLRP3 inflammasome. Furthermore, co-culturing neurons with SIRT3-overexpressing microglia resulted in reduced neuronal apoptosis and improved mitochondrial function, evidenced by lower ROS levels, alleviated mitochondrial depolarization and increased ATP production.
SIRT3 restrains pro-inflammatory microglia and NLRP3 inflammasome and alleviates neuroinflammation following HI brain injury.
我们旨在探讨沉默调节蛋白3（SIRT3）在改善缺氧缺血（HI）所致神经炎症中的作用。
建立HI大鼠模型，在构建HI模型前48小时，大鼠接受脑室内注射重组腺相关病毒9型载体。TTC和尼氏染色评估SIRT3对HI大鼠脑梗死和脑萎缩的影响。通过检测白细胞介素-1β（IL-1β）和髓过氧化物酶（MPO）阳性细胞评估神经炎症，并采用酶联免疫吸附测定法（ELISA）测定炎性细胞因子。还检测了离子钙接头蛋白1（IBA-1）、CD68和诱导型一氧化氮合酶（iNOS）阳性小胶质细胞以及NLRP3激活相关蛋白的表达。在氧糖剥夺（OGD）诱导的小胶质细胞模型中过表达SIRT3，检测细胞形态以及促炎细胞因子和NLRP3炎性小体激活相关蛋白的表达。此外，分析与过表达SIRT3的小胶质细胞共培养的神经元的线粒体损伤和凋亡情况。
SIRT3减轻了HI大鼠的脑梗死和萎缩。它还抑制神经炎症，减少IL-1β和MPO阳性细胞，并降低促炎细胞因子水平。在HI大鼠模型和OGD细胞模型中，SIRT3均抑制小胶质细胞和NLRP3炎性小体的过度激活。此外，与过表达SIRT3的小胶质细胞共培养的神经元凋亡减少，线粒体功能改善，表现为活性氧（ROS）水平降低、线粒体去极化减轻和三磷酸腺苷（ATP）生成增加。
SIRT3抑制促炎性小胶质细胞和NLRP3炎性小体，减轻HI脑损伤后的神经炎症。
