Liu Jiandong, Guo Xiangna, Yang Lu, Tao Tao, Cao Jun, Hong Zexuan, Zeng Fanning, Lu Yitian, Lin Chunshui, Qin Zaisheng
Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Department of Anesthesiology, The Affiliated Dongnan Hospital of Xiamen University, School of Medicine, Xiamen University, Zhangzhou, China.
Front Neurosci. 2022 May 23;16:889292. doi: 10.3389/fnins.2022.889292. eCollection 2022.
Celastrol plays a significant role in cerebral ischemia-reperfusion injury. Although previous studies have confirmed that celastrol post-treatment has a protective effect on ischemic stroke, the therapeutic effect of celastrol on ischemic stroke and the underlying molecular mechanism remain unclear. In the present study, focal transient cerebral ischemia was induced by transient middle cerebral artery occlusion (tMCAO) in mice and celastrol was administered immediately after reperfusion. We performed lncRNA and mRNA analysis in the ischemic hemisphere of adult mice with celastrol post-treatment through RNA-Sequencing (RNA-Seq). A total of 50 differentially expressed lncRNAs (DE lncRNAs) and 696 differentially expressed mRNAs (DE mRNAs) were identified between the sham and tMCAO group, and a total of 544 DE lncRNAs and 324 DE mRNAs were identified between the tMCAO and tMCAO + celastrol group. Bioinformatic analysis was done on the identified deregulated genes through gene ontology (GO) analysis, KEGG pathway analysis and network analysis. Pathway analysis indicated that inflammation-related signaling pathways played vital roles in the treatment of ischemic stroke by celastrol. Four DE lncRNAs and 5 DE mRNAs were selected for further validation by qRT-PCR in brain tissue, primary neurons, primary astrocytes, and BV2 cells. The results of qRT-PCR suggested that most of selected differentially expressed genes showed the same fold change patterns as those in RNA-Seq results. Our study suggests celastrol treatment can effectively reduce cerebral ischemia-reperfusion injury. The bioinformatics analysis of lnRNAs and mRNAs profiles in the ischemic hemisphere of adult mice provides a new perspective in the neuroprotective effects of celastrol, particularly with regards to ischemic stroke.
雷公藤红素在脑缺血再灌注损伤中发挥着重要作用。尽管先前的研究已经证实雷公藤红素治疗对缺血性中风具有保护作用,但其对缺血性中风的治疗效果及潜在分子机制仍不清楚。在本研究中,通过小鼠大脑中动脉短暂闭塞(tMCAO)诱导局灶性短暂性脑缺血,并在再灌注后立即给予雷公藤红素。我们通过RNA测序(RNA-Seq)对雷公藤红素治疗后的成年小鼠缺血半球进行lncRNA和mRNA分析。在假手术组和tMCAO组之间共鉴定出50个差异表达的lncRNA(DE lncRNA)和696个差异表达的mRNA(DE mRNA),在tMCAO组和tMCAO + 雷公藤红素组之间共鉴定出544个DE lncRNA和324个DE mRNA。通过基因本体(GO)分析、KEGG通路分析和网络分析对鉴定出的失调基因进行生物信息学分析。通路分析表明,炎症相关信号通路在雷公藤红素治疗缺血性中风中起关键作用。选择4个DE lncRNA和5个DE mRNA通过qRT-PCR在脑组织、原代神经元、原代星形胶质细胞和BV2细胞中进行进一步验证。qRT-PCR结果表明,大多数选定的差异表达基因显示出与RNA-Seq结果相同的倍数变化模式。我们的研究表明雷公藤红素治疗可有效减轻脑缺血再灌注损伤。对成年小鼠缺血半球lncRNA和mRNA谱的生物信息学分析为雷公藤红素的神经保护作用提供了新的视角,特别是在缺血性中风方面。