在缺氧缺血性脑损伤(HIBD)模型中已发现星形胶质细胞中NLR家族pyrin结构域3(NLRP3)炎症小体的激活。富含半胱氨酸的血管生成诱导剂61(CYR61)由反应性星形胶质细胞分泌。然而,CYR61对HIBD的作用及其相关机制尚不清楚.本研究旨在探讨CYR61在新生儿HIBD星形胶质细胞和NLRP3炎性体活化中的作用。在7天Sprague-Dawley大鼠幼崽中建立HIBD模型。进行神经行为评估和2,3,5-三苯基-氯化四唑染色。此外,大鼠原代星形胶质细胞通过氧糖剥夺/再灌注(OGD/R)建立体外HIBD细胞模型。然后,将慢病毒介导的CYR61过表达和sh-CYR61病毒转导到ODG/R处理的原代星形胶质细胞中。实时定量PCR检测相关基因的表达,westernblot,免疫荧光染色,和酶联免疫吸附测定。结果表明,缺氧缺血引起的短期神经功能缺损,神经元损伤,新生大鼠脑梗塞。在体内,HIBD模型中CYR61、NLRP3和胶质纤维酸性蛋白(GFAP)的表达上调。体外,CYR61高表达。CYR61过表达增加了GFAP和C3的表达,而降低了S100A10的表达。CYR61过表达增加了NLRP3、ASC、caspase-1p20和IL-1β。CYR61过表达通过促进IκBα和p65的磷酸化激活NF-κB。因此,CYR61参与新生儿HIBD进展,这可能与星形胶质细胞的激活有关,NLRP3炎性体,和NF-κB信号通路。
The activation of the NLR family pyrin domain containing 3 (NLRP3) inflammasome in astrocytes has been found in the hypoxic-ischemic brain damage (HIBD) model. Cysteine rich angiogenic inducer 61 (
CYR61) is secreted by reactive astrocytes. However, the effects of
CYR61 on HIBD and its related mechanisms remain unclear. This study sought to explore the role of CYR61 in the activation of astrocytes and the NLRP3 inflammasome in neonatal HIBD. HIBD models were established in 7-day Sprague-Dawley rat pups. Neurobehavioral evaluation and 2,3,5-triphenyl-tetrazolium chloride staining were performed. In addition, rat primary astrocytes were used to establish the cell model of HIBD in vitro by oxygen-glucose deprivation/reperfusion (OGD/R). Then,
CYR61-overexpression and sh-
CYR61 viruses mediated by lentivirus were transduced into ODG/R-treated primary astrocytes. The expressions of related genes were evaluated using real-time quantitative PCR, western blot, immunofluorescence staining, and Enzyme-linked immunosorbent assay. The results showed that hypoxia-ischemia induced short-term neurological deficits, neuronal damage, and cerebral infarction in neonatal rats. In vivo, the expressions of CYR61, NLRP3, and glial fibrillary acidic protein (GFAP) were up-regulated in the HIBD model. In vitro,
CYR61 exhibited high expression. CYR61 overexpression increased the expressions of GFAP and C3, whereas decreased S100A10 expression.
CYR61 overexpression increased the expression of NLRP3, ASC, caspase-1 p20 and IL-1β. CYR61 overexpression activated NF-κB by promoting the phosphorylation of IκBα and p65. Thus, CYR61 is involved in neonatal HIBD progress, which may be related to the activation of astrocytes, the NLRP3 inflammasome, and the NF-κB signaling pathway.