Department of Anesthesiology, The First Affiliated Hospital of Ningbo University, No. 59 Liuting Street, Haishu District, Ningbo, Zhejiang, China.
Department of Anesthesiology, The First Affiliated Hospital of Ningbo University, No. 59 Liuting Street, Haishu District, Ningbo, Zhejiang, China.
Gene. 2024 Jan 30;893:147888. doi: 10.1016/j.gene.2023.147888. Epub 2023 Oct 13.
Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS), acting as one common sepsis-associated organ injury, induces uncontrolled and self-amplifies pulmonary inflammation. Given the lack of clinically effective approaches, the mortality rate of it still remains high. Suramin(SUR), as an antiparasitic drug initially, was found to ameliorate sepsis associated ALI in our previous work. However, the underlying mechanism of its protective effects has not been clarified. Pyroptosis, categorized as an inflammatory form of programmed cell death, could aggravate lung inflammatory responses via inducing alveolar macrophages (AM) pyroptosis.
MH-S AM cell line was stimulated with or without lipopolysaccharide (LPS) or suramin, and the differential expression genes (DEGs) were excavated using RNA sequencing (RNA-seq). To identify the regulatory roles of these genes, pyroptosis-related genes (PRGs), GO/KEGG and GSEA analysis were conducted. We also performed WB, qRTPCR and ELISA to validate the RNA-seq results and further expound the protective effect of suramin.
624 DEGs were identified between control (CON) and lipopolysaccharide (LPS) groups, and enrichment analysis of these genes revealed significantly enriched pathways that related to immune system and signal transduction. Meanwhile, 500 DEGs were identified in LPS/SUR+LPS group. In addition to the pathways mentioned above, IL-17 pathway and C-type lectin receptor signaling pathway were also enriched. All 6 pathways were connected with pyroptosis. Concurrently, the "DESeq2" R package was used to identify differentially expressed PRGs. Nod1, Nod2, interleukin (IL)-1b, IL-6, tumor necrosis factor (TNF), NLRP3 were upregulated under LPS stimulation. Then, in SUR+LPS group, Nod2, IL-6, IL-1b, NLRP3 were downregulated. The validation results of WB, qRT-PCR, and ELISA showed: the protein and mRNA expression levels of NLRP3, caspase-1, GSDMD and the concentrations of IL-1b, IL-18 were decreased when treated with suramin and LPS.
Suramin could inhibit NLRP3/caspase-1/GSDMD canonical pyroptosis pathway in LPS-induced MH-S alveolar macrophages.
急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）作为一种常见的脓毒症相关器官损伤，可引起失控和自我放大的肺部炎症。由于缺乏临床有效的治疗方法，其死亡率仍然很高。苏拉明（SUR）最初是一种抗寄生虫药物，在我们之前的工作中发现可以改善脓毒症相关的 ALI。然而，其保护作用的潜在机制尚未阐明。细胞焦亡，作为一种程序性细胞死亡的炎症形式，可通过诱导肺泡巨噬细胞（AM）焦亡来加重肺炎症反应。
用脂多糖（LPS）或苏拉明刺激 MH-S AM 细胞系，使用 RNA 测序（RNA-seq）挖掘差异表达基因（DEGs）。为了确定这些基因的调节作用，进行了细胞焦亡相关基因（PRGs）、GO/KEGG 和 GSEA 分析。我们还进行了 WB、qRT-PCR 和 ELISA 验证 RNA-seq 结果，并进一步阐述苏拉明的保护作用。
在对照（CON）和脂多糖（LPS）组之间鉴定出 624 个 DEGs，这些基因的富集分析显示，与免疫系统和信号转导相关的途径显著富集。同时，在 LPS/SUR+LPS 组中鉴定出 500 个 DEGs。除了上述途径外，IL-17 途径和 C 型凝集素受体信号通路也有富集。所有 6 条途径都与细胞焦亡有关。同时，使用“DESeq2”R 包鉴定差异表达的 PRGs。Nod1、Nod2、白细胞介素（IL）-1b、IL-6、肿瘤坏死因子（TNF）、NLRP3 在 LPS 刺激下上调。然后，在 SUR+LPS 组中，Nod2、IL-6、IL-1b、NLRP3 下调。WB、qRT-PCR 和 ELISA 的验证结果表明，苏拉明和 LPS 处理后，NLRP3、caspase-1、GSDMD 的蛋白和 mRNA 表达水平以及 IL-1b、IL-18 的浓度降低。
苏拉明可抑制 LPS 诱导的 MH-S 肺泡巨噬细胞中 NLRP3/caspase-1/GSDMD 经典细胞焦亡途径。
