环状 RNA-Katnal1 通过 miR-31-5p/GSDMD 轴增强脓毒症诱导的肝损伤中的炎症性细胞焦亡。
circ-Katnal1 Enhances Inflammatory Pyroptosis in Sepsis-Induced Liver Injury through the miR-31-5p/GSDMD Axis.
机构信息
Department of Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001 Heilongjiang Province, China.
Department of Critical Care Medicine, The Sixth Affiliated Hospital of Harbin Medical University, Harbin, 150028 Heilongjiang Province, China.
出版信息
Mediators Inflamm. 2022 Aug 8;2022:8950130. doi: 10.1155/2022/8950130. eCollection 2022.
BACKGROUND
Sepsis is a systemic inflammatory response that can elicit organ dysfunction as well as circulatory diseases in serious cases. When inflammatory responses are especially dysregulated, severe complications can arise, including sepsis-induced liver injury. Various microRNAs along with circular (circ) RNAs are involved in inflammatory responses; nevertheless, their functions in regulating sepsis-induced liver injury remain unknown. The cecal ligation and puncture (CLP) procedure can induce liver injury as well as polymicrobial sepsis.
METHODS
In this study, CLP was used to induce liver injury as well as polymicrobial sepsis. Then, liver function, inflammatory cytokine expression, and hepatic histopathology were evaluated. High-throughput sequencing was employed to investigate the abnormal hepatic circRNA expression after CLP. Raw264.7 cells were utilized to simulation an sepsis inflammation model with LPS induce. The relative mRNA as well as protein levels of TNF-, IL-1, and IL-6 was explored by quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays. We explored functional connections among circRNAs, miR-31-5p, and gasdermin D (GSDMD) using dual-luciferase reporter assays. Western blot was employed to test GSDMD, caspase-1, and NLRP3 expression in mice and cell models.
RESULTS
Our results showed that CLP-induced sepsis promoted liver injury via increasing inflammatory pyroptosis. The abnormal expression of circ-Katnal1 played an important role in CLP-induced sepsis. Downregulating circ-Katnal1 suppressed LPS-induced inflammatory pyroptosis in Raw264.7 cells. Bioinformatics and luciferase reporter results confirmed that miR-31-5p and GSDMD were downstream targets of circ-Katnal1. Inhibiting miR-31-5p or upregulating GSDMD reversed the protective effects of silencing circ-Katnal1.
CONCLUSION
Taken together, circ-Katnal1 enhanced inflammatory pyroptosis in sepsis-induced liver injury through the miR-31-5p/GSDMD axis.
背景
败血症是一种全身炎症反应，在严重情况下可引起器官功能障碍和循环系统疾病。当炎症反应特别失调时，可能会出现严重并发症，包括败血症引起的肝损伤。各种 microRNA 和环状 (circ) RNA 参与炎症反应；然而，它们在调节败血症引起的肝损伤中的作用尚不清楚。盲肠结扎和穿刺 (CLP) 程序可诱导肝损伤和多微生物败血症。
方法
本研究采用 CLP 诱导肝损伤和多微生物败血症。然后，评估肝功能、炎症细胞因子表达和肝组织病理学。采用高通量测序技术研究 CLP 后肝组织 circRNA 表达的异常。使用 LPS 诱导的 Raw264.7 细胞模拟败血症炎症模型。通过定量聚合酶链反应 (PCR) 和酶联免疫吸附测定法研究 TNF-α、IL-1 和 IL-6 的相对 mRNA 和蛋白水平。我们通过双荧光素酶报告基因实验研究 circRNA、miR-31-5p 和 gasdermin D (GSDMD) 之间的功能联系。Western blot 检测小鼠和细胞模型中 GSDMD、caspase-1 和 NLRP3 的表达。
结果
我们的结果表明，CLP 诱导的败血症通过增加炎症细胞焦亡促进肝损伤。circ-Katnal1 的异常表达在 CLP 诱导的败血症中起重要作用。下调 circ-Katnal1 抑制了 LPS 诱导的 Raw264.7 细胞炎症细胞焦亡。生物信息学和荧光素酶报告结果证实，miR-31-5p 和 GSDMD 是 circ-Katnal1 的下游靶标。抑制 miR-31-5p 或上调 GSDMD 逆转了沉默 circ-Katnal1 的保护作用。
结论
综上所述，circ-Katnal1 通过 miR-31-5p/GSDMD 轴增强败血症诱导的肝损伤中的炎症细胞焦亡。
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