Sheng Ziyi, Song Hua, Gao Xianzhi, Shu Bian, You Yu, Liu Zuojin
Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing Medical University, Chongqing, 40010, China.
Inflammation. 2024 Nov 26. doi: 10.1007/s10753-024-02172-6.
This study aimed to investigate whether and how lipopolysaccharide (LPS) activated hepatic stellate cells (HSCs) regulate macrophage activity and to explore the impact of microRNAs (miRNAs) in exosomes from HSCs on this process. Mice subjected to LPS or cecal ligation and puncture (CLP) were used to explore sepsis-induced liver injury. Liver injury was evaluated using HE staining, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured. LPS-Exo or N-LPS-Exo from HSCs were added to hepatic macrophages, and iNOS, IL-1β, and TNF-α expression was detected via Western blotting. miRNA microarray analysis and PCR were used to evaluate differentially expressed miRNAs between LPS-Exo and N-LPS-Exo. Target genes were screened using the TargetScan database and verified with luciferase assays and WB. Inflammation and macrophage activity were observed in vivo using HE and CD86 staining in mice injected with PKH67-labeled LPS-Exo or N-LPS-Exo. Sepsis-related liver injury activates hepatic stellate cells, which regulate macrophage activity through exosomes. Specifically, exosomal miR-146a-5p secreted by hepatic stellate cells targets KLF-4, regulating the macrophage inflammatory response through the JNK signaling pathway. Exosomes containing miRNA-146a-5p released from HSCs following LPS treatment may increase macrophage sensitivity to LPS and trigger an inflammatory response. Exosomal miR-146a-5p derived from HSCs accelerates sepsis-induced liver injury by suppressing KLF-4 expression.
本研究旨在调查脂多糖（LPS）激活的肝星状细胞（HSCs）是否以及如何调节巨噬细胞活性，并探索HSCs来源的外泌体中的微小RNA（miRNAs）在此过程中的作用。使用接受LPS或盲肠结扎穿刺（CLP）的小鼠来探究脓毒症诱导的肝损伤。通过苏木精-伊红（HE）染色评估肝损伤，并测量丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平。将HSCs来源的LPS-Exo或N-LPS-Exo添加到肝巨噬细胞中，通过蛋白质免疫印迹法检测诱导型一氧化氮合酶（iNOS）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的表达。使用miRNA芯片分析和聚合酶链反应（PCR）评估LPS-Exo和N-LPS-Exo之间差异表达的miRNAs。使用TargetScan数据库筛选靶基因，并通过荧光素酶测定和蛋白质免疫印迹法进行验证。在注射PKH67标记的LPS-Exo或N-LPS-Exo的小鼠中，通过HE和CD86染色在体内观察炎症和巨噬细胞活性。脓毒症相关的肝损伤激活肝星状细胞，肝星状细胞通过外泌体调节巨噬细胞活性。具体而言，肝星状细胞分泌的外泌体miR-146a-5p靶向 Kruppel样因子4（KLF-4），通过JNK信号通路调节巨噬细胞炎症反应。LPS处理后HSCs释放的含有miR-146a-5p的外泌体可能会增加巨噬细胞对LPS的敏感性并引发炎症反应。HSCs来源的外泌体miR-146a-5p通过抑制KLF-4表达加速脓毒症诱导的肝损伤。
