Zhao Yangyang, Xiao Qiuxiang, Sun Tao, Yu Haiyun, Luo Muyun
The First Clinical Medical College, Gannan Medical University, Ganzhou City, Jiangxi Province, People's Republic of China.
Department of Pathology, The First Affiliated Hospital of Gannan Medical University, Ganzhou City, Jiangxi Province, People's Republic of China.
Neuropsychiatr Dis Treat. 2024 Jan 17;20:83-99. doi: 10.2147/NDT.S440065. eCollection 2024.
The aims of this study are to screen novel differentially expressed genes (DEGs) for intracerebral hemorrhage (ICH) and reveal the role of Lipocalin-2 (LCN2) in ICH.
We constructed the ICH model by injection of autologous whole blood into the right basal ganglia in rats. RNA-sequencing and bioinformatics analyses were performed to identify the DEGs between ICH and sham rats, and some important ones were confirmed using quantitative real-time PCR (qRT-PCR). LCN shRNA was used to knockdown of LCN2 in ICH rats. Pathological examination was carried out using 2,3,5-triphenyltetrazolium chloride (TTC) staining and Hematoxylin-eosin (HE) staining. Immunohistochemistry detected Caspase-3, and co-staining of Terminal dUTP nick end labeling (TUNEL) and NEUN staining were performed for neuron apoptosis assessment. Western blot analysis was performed to quantify pyroptosis-related proteins. Enzyme-linked immunosorbent assay (ELISA) was used to measure inflammatory cytokine levels.
ICH rats exhibited significant hematomas, higher brain water content, obvious interstitial edema, and inflammatory infiltration, as well as more apoptotic cells in brain tissues. RNA-seq analysis identified 103 upregulated and 81 downregulated DEGs. The expression of LCN2, HSPB1, CXCL10, and MEF2B were upregulated in ICH rats. ICH triggered the release of interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), and IL-18, and promoted the expression of pyroptosis-related proteins Caspase-1, GSDMD, NLRP3, and ASC. LCN2 knockdown attenuated the pathological characteristics of ICH, and also reduced pyroptosis in brain tissues.
Inhibition of LCN2 attenuates brain injury after ICH via suppressing pyroptosis, which provide guidance for ICH management.
本研究旨在筛选脑出血（ICH）新的差异表达基因（DEGs），并揭示脂质运载蛋白-2（LCN2）在ICH中的作用。
通过向大鼠右侧基底节注射自体全血构建ICH模型。进行RNA测序和生物信息学分析以鉴定ICH大鼠和假手术大鼠之间的DEGs，并使用定量实时PCR（qRT-PCR）确认一些重要的DEGs。LCN shRNA用于敲低ICH大鼠中的LCN2。使用2,3,5-三苯基氯化四氮唑（TTC）染色和苏木精-伊红（HE）染色进行病理检查。免疫组织化学检测Caspase-3，并进行末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）与NEUN染色的共染色以评估神经元凋亡。进行蛋白质免疫印迹分析以定量焦亡相关蛋白。酶联免疫吸附测定（ELISA）用于测量炎性细胞因子水平。
ICH大鼠表现出明显的血肿、更高的脑含水量、明显的间质水肿和炎性浸润，以及脑组织中更多的凋亡细胞。RNA测序分析鉴定出103个上调和81个下调的DEGs。LCN2、热休克蛋白B1（HSPB1）、CXC趋化因子配体10（CXCL10）和肌细胞增强因子2B（MEF2B）的表达在ICH大鼠中上调。ICH引发白细胞介素（IL）-1β、肿瘤坏死因子-α（TNF-α）和IL-18的释放，并促进焦亡相关蛋白Caspase-1、gasdermin D（GSDMD）、NLR家族含pyrin结构域蛋白（NLRP3）和凋亡相关斑点样蛋白（ASC）的表达。LCN2敲低减轻了ICH的病理特征，也减少了脑组织中的焦亡。
抑制LCN2通过抑制焦亡减轻ICH后的脑损伤，为ICH的治疗提供指导。
