Chen Yuanzhuo, Liu Dan, Li Xiaofeng, Nie Ke, Yan Songjuan, Peng Hu, Zhuang Yugang, Sun Mingming
Department of Emergency, Shanghai Tenth People's Hospital, School of Medicine Tongji University, Shanghai, 200072, China.
Department of Emergency, Shanghai Tenth People's Hospital, School of Medicine Tongji University, Shanghai, 200072, China.
Biochem Biophys Res Commun. 2023 May 7;655:90-96. doi: 10.1016/j.bbrc.2023.03.002. Epub 2023 Mar 7.
Acute lung injury (ALI) induced by sepsis and its complications have high morbidity and mortality rates globally. The objective of this study was to enhance our understanding of the underlying mechanism of ALI by identifying potential splicing events that are regulated in this condition.
The CLP mouse model was utilized for mRNA sequencing, and the expression and splicing data were analyzed. Verification of the changes in expression and splicing induced by CLP was conducted using qPCR and RT-PCR.
Our results showed that splicing-related genes were regulated, suggesting that splicing regulation may be a key mechanism in ALI. We also found that more than 2900 genes displayed alternative splicing in the lungs of mice with sepsis. Using RT-PCR, we verified that TLR4 and other genes had differential splicing isoforms in the lungs of mice with sepsis. We confirmed the presence of TLR4-s in the lungs of mice with sepsis using RNA-fluorescence in situ hybridization.
Our results suggest that sepsis-induced ALI can significantly alter splicing in the lungs of mice. The list of DASGs and splicing factors is valuable for further study in the search for new treatment approaches for sepsis-induced ALI.
由脓毒症及其并发症诱发的急性肺损伤(ALI)在全球范围内具有较高的发病率和死亡率。本研究的目的是通过识别在此病症中受到调控的潜在剪接事件,加深我们对ALI潜在机制的理解。
采用CLP小鼠模型进行mRNA测序,并对表达和剪接数据进行分析。使用qPCR和RT-PCR对CLP诱导的表达和剪接变化进行验证。
我们的结果表明剪接相关基因受到调控,提示剪接调控可能是ALI的关键机制。我们还发现,超过2900个基因在脓毒症小鼠的肺中表现出可变剪接。使用RT-PCR,我们验证了TLR4和其他基因在脓毒症小鼠的肺中有不同的剪接异构体。我们使用RNA荧光原位杂交证实了脓毒症小鼠肺中存在TLR4-s。
我们的结果表明,脓毒症诱导的ALI可显著改变小鼠肺中的剪接。差异可变剪接基因(DASG)和剪接因子列表对于进一步研究寻找脓毒症诱导的ALI的新治疗方法具有重要价值。