Exosomal microRNA-342-5p secreted from adipose-derived mesenchymal stem cells mitigates acute kidney injury in sepsis mice by inhibiting TLR9.

IF 3.7 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS Biological Procedures Online Pub Date : 2023-04-21 DOI:10.1186/s12575-023-00198-y

Wei Liu, Chenghuan Hu, Buyao Zhang, Mingxia Li, Fuxing Deng, Shuangping Zhao

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引用次数: 3

Abstract

Background: Sepsis-related acute kidney injury (AKI) is an inflammatory disease associated with extremely high mortality and health burden. This study explored the possibility of exosomes secreted by adipose-derived mesenchymal stem cells (AMSCs) serving as a carrier for microRNA (miR)-342-5p to alleviate sepsis-related AKI and investigated the possible mechanism.

Methods: Serum was obtained from 30 patients with sepsis-associated AKI and 30 healthy volunteers for the measurement of miR-342-5p, blood urea nitrogen (BUN), and serum creatinine (SCr) levels. For in vitro experiments, AMSCs were transfected with LV-miR-342-5p or LV-miR-67 to acquire miR-342-5p-modified AMSCs and miR-67-modified AMSCs, from which the exosomes (AMSC-Exo-342 and AMSC-Exo-67) were isolated. The human renal proximal tubular epithelial cell line HK-2 was induced by lipopolysaccharide (LPS) to construct a cellular model of sepsis. The expression of Toll-like receptor 9 (TLR9) was also detected in AKI cells and mouse models. The interaction between miR-342-5p and TLR9 was predicted by dual luciferase reporter gene assay.

Results: Detection on clinical serum samples showed that BUN, SCr, and TLR9 were elevated and miR-342-5p level was suppressed in the serum of patients with sepsis-associated AKI. Transfection with LV-miR-342-5p reinforced miR-342-5p expression in AMSCs and AMSC-secreted exosomes. miR-342-5p negatively targeted TLR9. LPS treatment enhanced TLR9 expression, reduced miR-342-5p levels, suppressed autophagy, and increased inflammation in HK-2 cells, while the opposite trends were observed in LPS-induced HK-2 cells exposed to AMSC-Exo-342, Rapa, miR-342-5p mimic, or si-TLR9. Additionally, the effects of AMSC-Exo-342 on autophagy and inflammation in LPS-induced cells could be weakened by 3-MA or pcDNA3.1-TLR9 treatment. Injection of AMSC-Exo-342 enhanced autophagy, mitigated kidney injury, suppressed inflammation, and reduced BUN and SCr levels in sepsis-related AKI mouse models.

Conclusion: miR-342-5p transferred by exosomes from miR-342-5p-modified AMSCs ameliorated AKI by inhibiting TLR9 to accelerate autophagy.

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脂肪源性间充质干细胞分泌的外泌体microRNA-342-5p通过抑制TLR9减轻脓毒症小鼠的急性肾损伤。

背景:败血症相关性急性肾损伤(AKI)是一种具有极高死亡率和健康负担的炎症性疾病。本研究探讨了脂肪源性间充质干细胞(AMSCs)分泌的外泌体作为microRNA (miR)-342-5p载体减轻脓毒症相关AKI的可能性，并探讨了其可能的机制。方法:采集30例败血症相关性AKI患者和30名健康志愿者的血清，测定miR-342-5p、血尿素氮(BUN)和血清肌酐(SCr)水平。在体外实验中，用LV-miR-342-5p或LV-miR-67转染AMSCs，获得mir -342-5p修饰的AMSCs和mir -67修饰的AMSCs，并从中分离外泌体(msc - exo -342和msc - exo -67)。采用脂多糖(LPS)诱导人肾近端小管上皮细胞系HK-2建立脓毒症细胞模型。在AKI细胞和小鼠模型中也检测到toll样受体9 (TLR9)的表达。通过双荧光素酶报告基因测定预测miR-342-5p与TLR9之间的相互作用。结果:临床血清样本检测显示败血症相关性AKI患者血清BUN、SCr、TLR9升高，miR-342-5p水平被抑制。转染LV-miR-342-5p可增强AMSCs和amsc分泌外泌体中miR-342-5p的表达。miR-342-5p负性靶向TLR9。LPS处理增强了TLR9表达，降低了miR-342-5p水平，抑制了自噬，并增加了HK-2细胞的炎症，而LPS诱导的HK-2细胞暴露于AMSC-Exo-342、Rapa、miR-342-5p mimic或si-TLR9时，观察到相反的趋势。此外，AMSC-Exo-342对lps诱导的细胞自噬和炎症的作用可以通过3-MA或pcDNA3.1-TLR9处理而减弱。在败血症相关AKI小鼠模型中，注射AMSC-Exo-342增强自噬，减轻肾损伤，抑制炎症，降低BUN和SCr水平。结论:miR-342-5p通过外泌体从miR-342-5p修饰的AMSCs中转移，通过抑制TLR9加速自噬来改善AKI。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biological Procedures Online 生物-生化研究方法

CiteScore

10.50

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0.00%

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审稿时长

>12 weeks

期刊介绍： iological Procedures Online publishes articles that improve access to techniques and methods in the medical and biological sciences. We are also interested in short but important research discoveries, such as new animal disease models. Topics of interest include, but are not limited to: Reports of new research techniques and applications of existing techniques Technical analyses of research techniques and published reports Validity analyses of research methods and approaches to judging the validity of research reports Application of common research methods Reviews of existing techniques Novel/important product information Biological Procedures Online places emphasis on multidisciplinary approaches that integrate methodologies from medicine, biology, chemistry, imaging, engineering, bioinformatics, computer science, and systems analysis.