转录组分析揭示了脓毒性心肌病中与细胞焦亡相关基因的作用机制。

Transcriptome analysis reveals the mechanism of pyroptosis-related genes in septic cardiomyopathy.

机构信息

Department of Cardiology, Zhongnan Hospital of Wuhan University, Wuhan, China.

Institute of Myocardial Injury and Repair, Wuhan University, Wuhan, China.

出版信息

PeerJ. 2023 Oct 19;11:e16214. doi: 10.7717/peerj.16214. eCollection 2023.

DOI:10.7717/peerj.16214

PMID:37872948

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10590578/

Abstract

BACKGROUND

Septic cardiomyopathy (SC) is characterized by myocardial dysfunction caused by sepsis and constitutes one of the serious complications of sepsis. Pyroptosis is a unique proinflammatory programmed cell death process. However, the role of pyroptosis in the development of SC remains unclear, and further study is required. The purpose of this study is to identify pyroptosis-related genes (PRGs) in SC and explore the mechanism of pyroptosis involved in the regulation of SC formation and progression.

METHODS

Differential expression analysis and enrichment analysis were performed on the SC-related dataset GSE79962 to identify differentially expressed genes (DEGs). PRGs were screened by intersecting genes associated with pyroptosis in previous studies with the DEGs obtained from GSE79962. The expression pattern of them was studied based on their raw expression data. Additionally, corresponding online databases were used to predict miRNAs, transcription factors (TFs) and therapeutic agents of PRGs. Lipopolysaccharide (LPS)-induced cell damage models in H9C2 and AC16 cell lines were constructed, cell activity was detected by CCK-8 and cell pyroptosis were detected by Hoechst33342/PI staining. Furthermore, these PRGs were verified in the external datasets (GSE53007 and GSE142615) and LPS-induced cell damage model. Finally, the effect of siRNA-mediated PRGs knockdown on the pyroptosis phenotype was examined.

RESULTS

A total of 1,206 DEGs were screened, consisting of 663 high-expressed genes and 543 low-expressed genes. Among them, ten PRGs (SOD2, GJA1, TIMP3, TAP1, TIMP1, NOD1, TP53, CPTP, CASP1 and SAT1) were identified, and they were mainly enriched in "Pyroptosis", "Ferroptosis", "Longevity regulating pathway", and "NOD-like receptor signaling pathway". A total of 147 miRNAs, 31 TFs and 13 therapeutic drugs were predicted targeting the PRGs. The expression trends of SOD2 were confirmed in both the external datasets and LPS-induced cell damage models. Knockdown of SOD2 induced increased pyroptosis in the AC16 LPS-induced cell damage model.

CONCLUSIONS

In this study, we demonstrated that SOD2 is highly expressed in both the SC and LPS-induced cell damage models. Knockdown of SOD2 led to a significant increase in pyroptosis in the AC16 LPS-induced cell damage model. These findings suggest that SOD2 may serve as a potential target for the diagnosis and treatment of SC.

摘要

背景

脓毒症性心肌病（SC）是由脓毒症引起的心肌功能障碍的特征，是脓毒症的严重并发症之一。细胞焦亡是一种独特的炎症程序性细胞死亡过程。然而，细胞焦亡在 SC 发展中的作用尚不清楚，需要进一步研究。本研究旨在鉴定 SC 中与细胞焦亡相关的基因（PRGs），并探讨涉及调节 SC 形成和进展的细胞焦亡的机制。

方法

对 SC 相关数据集 GSE79962 进行差异表达分析和富集分析，以鉴定差异表达基因（DEGs）。通过将先前研究中与细胞焦亡相关的基因与 GSE79962 中获得的 DEGs 进行交集，筛选 PRGs。根据其原始表达数据研究它们的表达模式。此外，还使用相应的在线数据库预测 PRGs 的 microRNAs、转录因子（TFs）和治疗剂。构建 H9C2 和 AC16 细胞系中脂多糖（LPS）诱导的细胞损伤模型，通过 CCK-8 检测细胞活性，通过 Hoechst33342/PI 染色检测细胞焦亡。进一步在外部数据集（GSE53007 和 GSE142615）和 LPS 诱导的细胞损伤模型中验证这些 PRGs。最后，通过 siRNA 介导的 PRGs 敲低检测对细胞焦亡表型的影响。

结果

共筛选出 1206 个 DEGs，其中包括 663 个高表达基因和 543 个低表达基因。其中，鉴定出 10 个 PRGs（SOD2、GJA1、TIMP3、TAP1、TIMP1、NOD1、TP53、CPTP、CASP1 和 SAT1），它们主要富集在“细胞焦亡”、“铁死亡”、“长寿调控途径”和“NOD 样受体信号通路”。总共预测到针对 PRGs 的 147 个 microRNAs、31 个 TFs 和 13 种治疗药物。SOD2 的表达趋势在外部数据集和 LPS 诱导的细胞损伤模型中均得到证实。在 AC16 LPS 诱导的细胞损伤模型中敲低 SOD2 导致细胞焦亡显著增加。