Sepsis plasma-derived exosomal miR-1-3p induces endothelial cell dysfunction by targeting SERP1.
Sepsis plasma-derived exosomal miR-1-3p induces endothelial cell dysfunction by targeting SERP1.
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DOI：
10.1042/cs20200573
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发表时间：
2021-01-29
期刊：
影响因子：
--
通讯作者：
Zhang X
中科院分区：
文献类型：
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作者：
Gao M;Yu T;Liu D;Shi Y;Yang P;Zhang J;Wang J;Liu Y;Zhang X
急性肺损伤（acute lung injury，ALI）是脓毒症患者死亡的主要原因。外泌体通过调节内皮细胞炎症反应、氧化应激和凋亡参与ALI的发生和发展，引起严重的肺血管渗漏和间质水肿。本研究探讨了脓毒症期间外泌体miRNAs对内皮细胞的影响。我们发现，在盲肠结扎穿孔（CLP）大鼠外泌体测序和脓毒症患者的外泌体，以及脂多糖（LPS）刺激的人脐静脉内皮细胞（HUVEC）中，miR-1- 3 p表达显著增加。然而，miR-1- 3 p在ALI中的具体生物学功能仍不清楚。因此，转染miR-1- 3 p的模拟物或抑制剂以调节其在HUVEC中的表达。采用CCK-8试剂盒、流式细胞仪、鬼笔环肽染色、Transwell法、乳酸脱氢酶（LDH）活性测定和Western blotting法检测细胞增殖、凋亡、收缩、通透性和膜损伤。利用miRNA相关数据库预测miR-1- 3 p靶基因，并通过荧光素酶报告基因进行验证。通过siRNA阻断靶基因的表达，探讨其作用机制。结果表明，在体内和体外，miR-1- 3 p表达增加，应激相关内质网蛋白1（SERP 1）表达减少。SERP 1是miR-1- 3 p的直接靶基因。上调的miR-1- 3 p通过靶向SERP 1抑制细胞增殖，促进细胞凋亡和细胞骨架收缩，增加单层内皮细胞通透性和膜损伤，导致内皮细胞功能障碍，削弱血管屏障功能，参与ALI的发生发展。miR-1- 3 p和SERP 1可能是脓毒症诱导的肺损伤的有希望的治疗候选者。
Acute lung injury (ALI) is the leading cause of death in sepsis patients. Exosomes participate in the occurrence and development of ALI by regulating endothelial cell inflammatory response, oxidative stress and apoptosis, causing serious pulmonary vascular leakage and interstitial edema. The current study investigated the effect of exosomal miRNAs on endothelial cells during sepsis. We found a significant increase in miR-1-3p expression in cecal ligation and puncture (CLP) rats exosomes sequencing and sepsis patients’ exosomes, and lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs) in vitro. However, the specific biological function of miR-1-3p in ALI remains unknown. Therefore, mimics or inhibitors of miR-1-3p were transfected to modulate its expression in HUVECs. Cell proliferation, apoptosis, contraction, permeability, and membrane injury were examined via cell counting kit-8 (CCK-8), flow cytometry, phalloidin staining, Transwell assay, lactate dehydrogenase (LDH) activity, and Western blotting. The miR-1-3p target gene was predicted with miRNA-related databases and validated by luciferase reporter. Target gene expression was blocked by siRNA to explore the underlying mechanisms. The results illustrated increased miR-1-3p and decreased stress-associated endoplasmic reticulum protein 1 (SERP1) expression both in vivo and in vitro. SERP1 was a direct target gene of miR-1-3p. Up-regulated miR-1-3p inhibits cell proliferation, promotes apoptosis and cytoskeleton contraction, increases monolayer endothelial cell permeability and membrane injury by targeting SERP1, which leads to dysfunction of endothelial cells and weakens vascular barrier function involved in the development of ALI. MiR-1-3p and SERP1 may be promising therapeutic candidates for sepsis-induced lung injury.